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.

OBJECTIVEUsing molecular epidemiology methods to investigate relationship between genotypes and drug-resistance of neisseria (N.) gonorrhoeae in Shanghai area.

METHODSA random amplified polymorphic DNA (RAPD) fingerprint method at the molecular level was used to differentiate the strains which were isolated from the outpatients of sexually transmitted disease clinics. The sensitivity to antibiotic of the 78 N. gonorrhoeae strains on 9 different antibiotics was tested and the relationship between different genotypes and phenotypes was studied.

RESULTSSelected RAPD primer could give out a group of amplification polymerase chain reaction bands with some main segments common to all the N. gonorrhoeae strains tested and some segments were different among the N. gonorrhoeae strains. All the 78 N. gonorrhoeae strains could be classified as three different groups (I, II and III). The strains could also be distinguished as four types (A, B, C and D) according to drug-resistance status. Using correspondence analysis method, the relationship between the three genotypes and four resistance types could be identified.

CONCLUSIONRAPD fingerprint seemed a useful genotyping method and could be used for molecular epidemiological studies.

Adolescent ; Adult ; Anti-Bacterial Agents ; pharmacology ; China ; epidemiology ; DNA Fingerprinting ; DNA, Bacterial ; genetics ; Drug Resistance, Bacterial ; Female ; Genotype ; Gonorrhea ; epidemiology ; microbiology ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Molecular Epidemiology ; Neisseria gonorrhoeae ; classification ; drug effects ; genetics ; Random Amplified Polymorphic DNA Technique

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

OBJECTIVETo observe the maintenance effect of polyglycosides of Tripterygium wilfordii (GTW) on remission in postoperative Crohn disease (CD).

METHODSFrom 2005 to 2007, 45 adult cases of postoperative Crohn disease were randomly divided into two groups, GTW group and mesalazine group, which received GTW and mesalazine treatment respectively. CD activity index (CDAI) and clinical markers were collected at 0, 3, 6, 12 months or at the onset of symptoms. Ileocolonoscopy was performed at the end of the trial (1 year after operation) or at the onset of symptoms, and recurrence score were recorded.

RESULTSNo clinical recurrence was ascertained in both groups at 3 months. Four patients (18.2%) in GTW group relapsed and 5 (21.7%) in mesalazine group relapsed at 6 months (P=0.530). Seven patients (31.8%) in GTW group and 9 (39.1%) in mesalazine group relapsed at one year (P=0.421). Ten patients (45.5%) in GTW group had endoscopic recurrence compared with 14 (60.9%) in mesalazine group at one year(P=0.231). There were no significant differences between two groups.

CONCLUSIONGTW is similar to mesalazine in maintenance of remission of postoperative Crohn disease.

Adolescent ; Adult ; Aged ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Crohn Disease ; drug therapy ; Glycosides ; therapeutic use ; Humans ; Male ; Mesalamine ; therapeutic use ; Middle Aged ; Phytotherapy ; Postoperative Period ; Recurrence ; Treatment Outcome ; Tripterygium ; chemistry ; Young Adult

OBJECTIVETo investigate risks and clinical effects of operative treatment for cervical vertebral fracture and dislocation associated with unilateral vertebral artery injury.

METHODSThis group consisted of 76 cases of closed cervical spine trauma combined with unilateral vertebral artery injury (23 cases of bilateral facet dislocation, 28 unilateral facet dislocation and 25 fracture). All patients underwent prospective examination of cervical spine MRI and vertebral artery two-dimensional time-of-flight (2D TOF) magnetic resonance angiography (MRA), and anterior cervical decompression. The healthy vertebral artery paths were evaluated before the surgery, and were protected during the surgery according to the anatomical signs.

RESULTSThere were no acute or chronic clinical damage symptoms in 76 cases after surgery. No neural damage symptoms were observed in patients with normal neural functions. The neural functions of incomplete paralyzed patients were improved in different grades.

CONCLUSIONSReliable anterior operation can produce good results for cervical fracture and dislocation with unilateral vertebral artery injury. Detecting the course of uninjured vertebral artery before operation and locating the anatomical site during operation are effective to avoid damaging vertebral artery of uninjured side.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cervical Vertebrae ; injuries ; Child ; Female ; Humans ; Joint Dislocations ; surgery ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Vertebral Artery ; injuries ; Wounds, Nonpenetrating ; surgery