AIM To study the effect of Vit E on the MLCK activity and expression in the liver of atherosclerosis model rabbits. METHODS The MLCK activity of rabbit liver was measured by the method of ?- 32P incorporated and its expression was detected by immunofluorescent. RESULTS The model of atherosclerosis was estabilished. After rabbits were fed with cholesterol for four weeks and twelve weeks, the activity of MLCK increased markedly, and there was significantly statistical difference compared with the normal control (P0.05). MLCK expression increased after the rabbits was fed with cholesterol for four weeks, and this increase became more obvious had been the rabbits was fed with cholesterol for twelve weeks. The expression decreased when vitamin E had been added into the cholesterol fed. CONCLUSION The pathology of liver may be associated with the increase of the activity of MLCK. Vit E may reduce MLCK activity and protect hepatocyte from injury.

Objective:To establish a method for measuring the activity of cornea epithelium quantitatively. Methods:Rabbit corneas were burnt either by alkali or by CO2 laser. The lamellar cornea was cut at the end of 1,2 and 3 weeks and cultured in 2 ml DMEM with 5% CO2, 37℃ for 1 h.Then 200 μl of MTT was added to the culture followed by incubation for another 4 h. The supernatant was discarded and 4 ml of DMSO was added into each culturedish for dissolving MTT completely under the condition of room temperature.200 μl of DMSO sample was added to each well of 96-well plate and each sample was triplicated. The absorbance of the plate was measured at 490 nm ultraviolet. Results:The D value of the burnt corneas was obviously lower than that of the normal ones(P<0.01). Conclusion:MTT method can be used to measure the activity of cornea epithelium quantitatively.

Depression is a common emotional disorder that seriously affects people's life and health all over the world. The pathogenesis of depression is complex, and traditional Chinese medicine (TCM) for antidepressants has a good therapeutic effect because of its multi-component, multi-pathway, and multi-target action mode. At present, the anti-depressive mechanism of TCM has not been fully clarified, but it is clear that depression is closely related to metabolic health. Therefore, in order to further explore the anti-depressive mechanism of TCM, this paper proposes research strategies on the anti-depressive mechanism of TCM based on functional metabolomics from the perspective of metabolism, the potential biomarkers of depression are analyzed with the help of multi-omics combined analysis technology, and the functional molecules of TCM for antidepressant are studied. Molecular biology techniques are used to accurately capture the molecular interactions between biomarkers of depression and functional compounds, which identify effective drug targets and further elucidate the biochemical functions and related mechanisms involved in depression metabolic disorders. This paper systematically reviews the research strategies and applications of functional metabolomics in the anti-depressive mechanisms of TCM, expounds on the core value of functional metabolomics, and summarizes the current research status and hot issues of TCM for antidepressants in recent years, providing new methods and new ideas for the study of mechanisms of TCM with the help of functional metabolomics.

Humans ; Siblings ; Forensic Medicine ; Forensic Genetics

OBJECTIVETo investigate whether GM-CSF can enhance the antiviral effect of HBsAg DNA vaccine.

METHODSDivided animals into 8 groups. Group A: pcDNA3.1-S 100microg; Group B: pcDNA3.1-GM-CSF-S 100microg; Group C: pcDNA3.1-S-GM-CSF 100microg; Group D: pcDNA3.1-S 50microg + pcDNA3.1-GM-CSF 50microg; Group E: pcDNA3.1-GM-CSF 100microg; Group F: recombinant HBsAg vaccine 1microg; Group G: pcDNA3.1,100microg; Group H: PBS 100microl. Serum HBsAg level and concentration of IL-2, IL-4 and IFN-gamma were examined using commercial ELISA kit. The [3H] thymidine incorporation into DNA of Spleen cells was measured; HBsAg expression of hepatocytes from HBV-transgenic mice was assessed using immunohistochemical analysis.

RESULTSSerum HBsAg level was lower and concentration of IL-2, IFN-gamma and SI was higher in mice immunized with pcDNA3.1-GM-CSF-S than those from mice immunized with pcDNA3.1-S and other groups (F=11.262, P<0.01, F=8.147, P<0.01, F=62.275, P<0.01, F=116.28, P<0.01. Less Hepatic HBsAg expression and decline of pcDNA3.1-GM-CSF-S of mice immunized with pcDNA3 were observed in comparison with control groups (F=41.439, P<0.01). Liver histological analysis showed no evidence of necrosis or inflammation in various groups.

CONCLUSIONThe plasmid co expressing GM-CSF and HBsAg could improve HBsAg-specific humoral and cellular immune responses induced by plasmid encoding HBsAg alone and enhance HBsAg DNA vaccine antivirus effect.

Animals ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B Vaccines ; immunology ; Interferon-gamma ; biosynthesis ; Interleukin-2 ; biosynthesis ; Interleukin-4 ; biosynthesis ; Lymphocyte Activation ; Male ; Mice ; Mice, Transgenic ; Plasmids ; Vaccines, DNA ; immunology

Objective To detect the mutations of ATP2C1 gene in patients with Hailey-Hailey dis- ease (HHD).Methods PCR and direct sequencing were performed in 17 patients and 120 healthy controls to screen the mutations in the exons of ATP2C1 gene.Results Eight mutations were identified in nine probands, including three deletion mutations (nt1464-1487 del/nt1462-1485del,1523delAT,2375delTTGT),three splice site mutations (360—2A→G,1415—2A→T,2243+2T→C) and two missence mutations (C920T and G1942T).None of the above mutations was found in the controls.Conclusion Eight specific novel mutations were identified in nine probands of HHD,which could be causative factors of the disease.

OBJECTIVETo investigate the association of angiotensin-converting enzyme 2 (ACE2) gene polymorphisms and coronary artery disease (CAD) in patients with type 2 diabetes mellitus (T2DM).

METHODSThis study involved 121 patients with T2DM and 94 with diabetic macroangiopathy. The polymorphisms of G8790A in ACE2 gene was analyzed using PCR-restriction fragment length polymorphism analysis in these patients, and the clinical, biochemical and echocardiographic data were also analyzed.

RESULTSNo obvious difference was found in the genotyping data between the two groups. Among the male patients with diabetic macroangiopathy, the interventricular septal end-diastolic thickness (IVSTd) were significantly greater in patients of GG genotypes of ACE2 gene G8790A than in those of AA genotypes (P<0.01), and the left ventricular mass (LVMI) and urine protein were also significantly higher in GG genotypes (P<0.05). No similar results were found the uncomplicated diabetic group or the female diabetic patients with CAD.

CONCLUSIONThe ACE2 gene G8790A polymorphism plays a role in the pathogenesis of CAD in patients with type 2 diabetes, suggesting that ACE2 genotyping is helpful to screen the susceptible patients.

Adult ; Aged ; Alleles ; Base Sequence ; Case-Control Studies ; Coronary Disease ; complications ; genetics ; Diabetes Mellitus, Type 2 ; complications ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Peptidyl-Dipeptidase A ; genetics ; Polymorphism, Genetic ; Sequence Analysis, DNA

OBJECTIVETo analysis the risk factors predicting intracardial thrombus after prosthetic valve replacement.

METHODSThe clinical data of 29 cases from January 2005 to April 2009 with intracardial thrombus after prosthetic valve replacement during a 1-year follow-up was retrospectively analyzed. There were 11 male and 18 female, aged from 12 to 70 years with a mean of 48 years. The risk factors of intracardial thrombus were examined by univariate and multivariate analysis.

RESULTSUnivariate analysis found that bioprosthetic valve replacement, anticoagulation using aspirin, valve replacement at mitral position, atrial fibrillation, preoperative and postoperative internal diameter of left atrium, postoperative fibrinogen were predict factors of intracardial thrombus after prosthetic valve replacement (P < 0.05). Logistic regression analysis showed valve replacement at mitral position (OR = 9.815, P < 0.05), atrial fibrillation (OR = 5.267, P < 0.05), preoperative internal diameter of left atrium (OR = 4.529, P < 0.05) were significant risk factors of intracardial thrombus after prosthetic valve replacement.

CONCLUSIONSValve replacement at mitral position, atrial fibrillation, and preoperative internal diameter of left atrium are the correlated risk factors of intracardial thrombus after prosthetic valve replacement. Anticoagulation after prosthetic valve (especially bioprosthetic valve) replacement should be standardized to prevent intracardial thrombus formation.

Adolescent ; Adult ; Aged ; Child ; Female ; Follow-Up Studies ; Heart Diseases ; etiology ; Heart Valve Prosthesis Implantation ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Retrospective Studies ; Risk Factors ; Thrombosis ; etiology ; Young Adult

OBJECTIVETo analysis the causes of valve prosthesis-patient mismatch (PPM) after mitral valve replacement in Chinese patients.

METHODSConsecutive 100 patients for elective mitral valve replacement from January 2009 to June 2009 were enrolled and followed for this study. There were 37 males and 63 females. The mean age at operation was (52 ± 9) years (ranging 32 to 76 years). The predominant mitral valve lesion was stenosis in 60 patients, regurgitation in 14 patients and mixed in 26 patients. Among them, 63 patients were combined tricuspid valve regurgitation. Mitral valve effective orifice area was measured by Doppler echocardiography in 100 patients who received mitral valve replacement and indexed for body surface area (EOAI). PPM was defined as not clinically significant if the EOAI was above 1.2 cm(2)/m(2), as moderate if it was >0.9 and ≤ 1.2 cm(2)/m(2), and as severe if it was ≤ 0.9 cm(2)/m(2). By using the criteria, all 100 patients were classified to two groups: PPM group and no PPM group. The clinical characteristic of the patients between the two groups was compared to determine the causes of PPM and the predictors of outcomes after mitral valve replacement, such as the gender, age, valve prosthesis type, size, body surface area, and mitral valve lesion, et al.

RESULTSOf the 100 patients after MVR, 52 (52.0%) had significant PPM, 51 (51.0%) had moderate PPM, and 1 (1.0%) had severe PPM. In comparison to patients in no PPM group, patients in PPM group had a significantly larger body surface area [(1.76 ± 0.17) m(2) vs. (1.59 ± 0.13) m(2), P < 0.01] and higher prevalence of male gender (55.8% vs. 16.6%, P < 0.01). The other preoperative and operative data were similar in both groups, such as the valve prosthesis type, size, and mitral valve lesion, et al. There were no significant differences in postoperative Doppler-echocardiographic data of cardiac structure and heart function between the two groups (P > 0.05).

CONCLUSIONSThe higher incidence of PPM in mitral valve position was in male or large body surface area patients. At the time of operation, surgeons should consider the related factors, such as the patient's gender and body surface area, et al. A larger prosthesis size might be implanted to avoid PPM in mitral valve position.

Adult ; Aged ; Female ; Heart Valve Prosthesis ; adverse effects ; Heart Valve Prosthesis Implantation ; adverse effects ; methods ; Humans ; Male ; Middle Aged ; Mitral Valve ; surgery

OBJECTIVETo set up random amplified polymorphic DNAs (RAPD) method in genotyping Neisseria gonorrhoeae on DNA level, and to explore its use to trace the source of infection.

METHODSFour different pretreatments were used to extract the Neisseria gonorrhoeae genomic DNA with its advantages and disadvantages compared. Arbitrary sequence was then used to amplify the genomic DNA of Neisseria gonorrhoeae and RAPD fingerprint maps was applied to distinct the Neisseria gonorrhoeae strains. Finally, RAPD fingerprint of Neisseria gonorrhoeae strain between patient and his/her sexual partner was compared.

RESULTSCetyltrimethylammonium bromide method was classical in extracting genomic DNA, and could get integrated genomic DNA and good fingerprint maps, since main segments were common to all the Neisseria gonorrhoeae but some were different among strains so that the fingerprint of different Neisseria gonorrhoeae were distinctive. However, fingerprint maps of Neisseria gonorrhoeae collected from sex partners were quite similar.

CONCLUSIONBased on genomic levels, effective fingerprint maps could be identified and to classify the Neisseria gonorrhoeae into different genotypes. RAPD fingerprint maps could be used to trace the source of infection.

DNA Fingerprinting ; DNA, Bacterial ; analysis ; Genotype ; Humans ; Neisseria gonorrhoeae ; classification ; genetics ; Random Amplified Polymorphic DNA Technique