AIM: To investigate the influence and mechanism of recombinant macrophage migration inhibitory factor(rMIF) on fibroblasts.METHODS: MRC-5 fibroblasts were divided into two groups: the treated group was treated with rMIF(25-100 ?g/L,12 h,24 h or 48 h) and the control was non-rMIF treatment.The activity of proliferation in both groups was investigated and compared by CCK-8 means.Synthesis of collagen in the culture supernatants was detected by the hydroxyproline.The expression of collagen type I mRNA was examined using RT-PCR analysis.The level of collagen type I protein induced by rMIF was quantified by Western blotting.RESULTS: The production of proliferation ratio of fibroblasts treated with 50 ?g/L and 100 ?g/L rMIF at 24 h or 48 h were increased obviously(P

we first find that the protoplast of Xanthomonas campestris can synthesize and secret Xanthan in the high permeable nutrition containing sucrose as substrate.

Microbiology is an important,fundamental and obligatory course in contemporary life science.This article introduces that teaching group of microbiology in Nankai University realizes transformation of teaching center,fully embodies the modernization of teaching notion and gives full play to students' main effect practically by adhering to teaching reform as center,optimizing teaching method as measure,communicating in and after class and using multi-media and teaching web.Therefore,teaching system is established to adapt to modern teaching notion and eventually microbiology course becomes a cultivation platform to foster elites with both solid fundamental theory and innovating mind.

Gamma-linolenic acid (GLA, C18:3delta6.9.12) is nutritional and important polyunsaturated fatty acid in human and animal diets. GLA play an important role in hormone regulation and fatty acid metabolization. Furthermore it is also the biological precursor of a group of molecules, including prostaglandins, leukotrienes and thromboxanes. Vast majority of oilseed crops do not produce GLA, but linoleic acid (LA, C18:2delta9.12) as its substrate. GLA is only produced by a small number of oilseed plants such as evening promrose ( Oenotheera spp.), borage (Borago officinalis) and etc. delta6-fatty acid desaturase (D6D) is the rate-limiting enzyme in the production of GLA. It can convert from linoleic acid to linolenic acid. To produce GLA in tobacco, plant expression vector was first constructed. To facilitate preparation of plant expression constructs, flanking Xba I and Bgl II restriction enzyme sites were added to the coding region of clone pTMICL6 by PCR amplification. pTMICL6 contains delta6-fatty acid desaturase gene cloned from Mortierella isabellina which is an oil-producing fugus. The PCR product was purified and subcloned into the plant expression vector pGA643 to generate the recombinant vector pGAMICL6 which contains the ORF of the D6D gene of Mortierella isabellina, together with regulatory elements consisting of the cauliflower mosaic virus 35S promoter and the nopaline synthase (nos) termination sequence. The plasmid pGAMICL6 was transformed into Agrobacterium tumefaciens strain LBA4404 by method of freeze thawing of liquid nitrogen. Transformants were selected by plating on YEB medium plates containing kanamycin and streptomycin and grown overnight at 28 degrees C, then transformants were further identified by PCR. The positive transformant containing the plant expression vector pGAMICL6 was transformed into tobacco ( Nicotiana tabacum cv. Xanthi) via Agrobacterium infection. Transgenic plants were selected on 100 microg/mL kanamycin. Plants were maintained in axionic culture under controlled conditions. Total nucleic acids were extracted and purified from anti-kanamycin transgenic tobacco and were analysed by PCR. 48 out of 80 transgenic plants were positive, in other words, transformation efficiency is 60% . This shows that Mortierella isabellina D6D gene is transformed into tobacco. Genomic DNA from PCR positive transgenic tobacco plants was digested with Hind III restriction enzyme and fractionated by agarose gel electrophoresis. Southern blotting was performed with strandard procedures for vacuum transfer of nucleic acids to nylon membrane. The probe was delta6-fatty acid desaturase gene from M. isabellina, which was labeled with DIG-dUTP via random-primed labeling. Hybridization and immumological detection were carried out the kit of DIG detection. The result shows single hybridizing bands in each of the transgenic tobacco plants DNA, but no hybridization was observed to non-transgenic tobacco. This indicates that delta6-fatty acid desaturase gene is integrated into the genome of transgenic tobacco. To provide further evidence that the introduction of the M. isabellina cDNA into the tobacco genome was responsible for the novel desaturation products, total RNA was isolated from GLA-positive transgenic tobacco plants via both PCR and Southern blotting and separated by electrophoresis through 1% formaldehyde agarose gel. Northern blotting including probe labeling, hybridization and detection was the same as Southern blotting in operation approach. A positive hybridization signal of identical mobility was obtained from RNA isolated from the transgenic tobacco plants, but not from the control tobacco plant. At last, total fatty acids extracted from the positive transgenic tobacco were analyzed by gas chromatography (GC) of methyl esters to confirm the transgenic tobacco containing a functional delta6-fatty acid desaturase gene. The result shows that two peaks were observed in the chromatogram of FAMes. GLA and octadecatetraenoic acid (OTA, C18:4delta6.9.12.15) respectively have 19.7% and 3.5% of the total fatty acids in the transgenic plant. The presence of both GLA and OTA indicates that the delta6-fatty acid desaturase used both linoleic acid and a-linolenic acid (ALA, C18:3delta6.9.12.15) as substrates, and this may be responsible for the decrease in ALA observed in the transgenic line. That was the first report about the expression of M. isabellina delta6-fatty acid desaturase gene in tobacco. All results mentioned above have laid the foundation of the thorough studying on an breeding transgenic oilseeds containing GLA to change the fatty acid composition of conventional oilseeds, it is significant to study on regulation mechanism of fatty acid desaturase.
Agrobacterium tumefaciens ; genetics ; Fatty Acid Desaturases ; genetics ; metabolism ; Genetic Vectors ; genetics ; Mortierella ; enzymology ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Polymerase Chain Reaction ; Tobacco ; genetics ; metabolism ; gamma-Linolenic Acid ; biosynthesis

Objective To describe the prevalence of deliberate self-harm (DSH) and suicidal behaviors (SIB) as well as the relationship between them in a college student population.Methods A total of 4063 medical students were selected under the cluster sampling method in Anhui province.Data were analyzed by Pearson Chi-square and logistic regression.Results A total of 4063 ( 13.4％ )students reported that they had deliberately harmed themselves during the past 12 months.The acts of DSH with 1 and more than or equal to 2 times occurrence accounted for 3.7％ and 9.6％ among all the respondents.The act of DSH was significantly higher among boys than that among girls.Rates of suicide ideation,suicide plan,attempted suicide and SIB in the last year were 4.5％,1.4％,0.6％ and 4.9％,respectively.However no statistically significant difference was found in different sex.Students in the DSH group that with SIB were found to have lower positive coping levels (OR=0.5,95％CI:0.3-0.8),getting less support from fricnds (3-5 vs.less than or equal to 2:OR=0.6,95％CI:0.3-0.9;more than or equal to 6 vs.less than or equal to 2:OR=0A,95％CI:0.2-0.8),with higher negative coping levels (OR=2.1,95％CI:1.2-3.7),having more serious depressive symptoms (OR=2.9,95％ CI:1.6-5.2) and anxiety symptoms (OR=2.2,95％ CI:1.2-3.8),having more serious sleeping problems (OR=1.7,95％CI:1.1-2.8 ) and perceived fat (fat vs.moderate:OR=2.0,95％CI:1.1-3.6) than the DSH group without SIB.The rates of SIB in students with DSH behavior were significantly higher than those in students without those behaviors (OR=4.7,95％ CI:3.5-6.4).Psyehosocial variables could attenuate the relationship between the DSH status and suicidal events (OR=3.3,95％CI:2.4-4.5).The DSH frequency exhibited a curvilinear relationship to SIB (OR=3.1-10.0) and psychosocial variables also attenuated this relationship (OR=2.4-5.9).Conclusion It was well known that SIB was not a suicidal gesture but our findings suggested that the presence of DSH might trigger the suicidal attempts.

OBJECTIVETo study the accumulation of fluoride in rat hippocampus and its effect on cholinesterase activity.

METHODSRats were subchronically exposed to NaF, and fluoride concentration and cholinesterase activity in rat hippocampus were determined.

RESULTSFluoride concentration in rat hippocampus was significantly correlated with the dosage of fluoride, and there were significant differences among high dosage group [(13.03 +/- 1.79) micro g/g], low dosage group [(9.83 +/- 0.92) micro g/g] and control [(8.27 +/- 1.11) micro g/g], P < 0.01. Acetylcholinesterase activities among three groups [(0.111 +/- 0.031) micro mol/mg, (0.143 +/- 0.025) micro mol/mg, (0.183 +/- 0.027) micro mol/mg] were also significantly different (P < 0.01), which was negatively correlated with fluoride concentration in rat hippocampus (r = -0.700, P < 0.01). The activity of butylcholinesterase in high dosage group [(0.041 +/- 0.010) micro mol/mg] was different from that of control [(0.067 +/- 0.025) micro mol/mg, P < 0.05], but the activity was not significantly related with fluoride concentration in rat hippocampus (r = -0.317, P = 0.094).

CONCLUSIONFluoride may go through the blood-brain barrier and accumulate in rat hippocampus, and inhibit the activity of cholinesterase.

Acetylcholinesterase ; metabolism ; Animals ; Blood-Brain Barrier ; Butyrylcholinesterase ; metabolism ; Fluoride Poisoning ; metabolism ; Fluorides ; pharmacokinetics ; Hippocampus ; metabolism ; Male ; Organ Size ; Rats ; Rats, Sprague-Dawley

Design space approach is applied in this study to enhance the robustness of first ethanol precipitation process of Codonopsis Radix (Dangshen) by optimizing parameters. Total flavonoid recovery, dry matter removal, and pigment removal were defined as the process critical quality attributes (CQAs). Plackett-Burman designed experiments were carried out to find the critical process parameters (CPPs). Dry matter content of concentrated extract (DMCE), mass ratio of ethanol to concentrated extract (E/C ratio) and concentration of ethanol (CEA) were identified as the CPPs. Box-Behnken designed experiments were performed to establish the quantitative models between CPPs and CQAs. Probability based design space was obtained and verified using Monte-Carlo simulation method. According to the verification results, the robustness of first ethanol precipitation process of Dangshen can be guaranteed by operating within the design space parameters. Recommended normal operation space are as follows: dry matter content of concentrated extract of 45.0% - 48.0%, E/C ratio of 2.48-2.80 g x g(-1), and the concentration of ethanol of 92.0% - 92.7%.
Chemical Precipitation ; Chemistry, Pharmaceutical ; methods ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification

Objective To investigate the predietors of road traffic accidents(RTAs)in middle school students.a ease-control study WaS designed to seek new clues for prevention and control on RTAs in this population.Methods Cases were 116 students who experienced RTAs from 2005/01/01 to 2005/12/31.116 students were.served as controls who were from the same clams of the cases and matched by age and sex.Information on personal behavior,family conditions,and knowledge,attitude,practice(KAB)on road traffic of these children were collected and compared.Conditionallogistic regressions were conducted to analyze the relationship between these factors and RTAs.Results Compared to the controls.the cases showed more negative attitude and risk-taking-behaviors especially in riding bicycle.However,knowledge on road safety showed less significant differences between eaSe and control groups.The negative attitude and risk-taking-behaviors were independent risk factors for RTAs by logistic regression.In addition.distance from home to school that longer than 1000 m seemed a risk factors for RTAs.while high education levels among mothers WaS a protective factor.Conclusion Road safety attitude and behavior.distance from home to school were the main influenee factors for RTAs among middle school students.It seemed more important in improving the attitude and behavior on road safety than to enhance the knowledge for prevention and eontrol of RTAs among middle school students.

Adult ; China ; epidemiology ; Circoviridae Infections ; epidemiology ; virology ; DNA, Viral ; analysis ; Female ; Genotype ; Hepatitis, Viral, Human ; virology ; Humans ; Male ; Substance Abuse, Intravenous ; virology ; Torque teno virus ; genetics ; isolation & purification

OBJECTIVETo evaluate the vascularized (deep circumflex iliac vessels) iliac crest graft with internal oblique muscle as a method for reconstruction of composite mandibular defect.

METHODSVascularized iliac crest graft with internal oblique muscle was used to reconstruct the composite mandibular defects in 10 patients. All clinical data were analyzed retrospectively. A detailed inspection of the case-notes was undertaken to ascertain the presenting diagnosis, the surgery, the complications and the outcome. The type of mandibular defect was recorded.

RESULTSOf the 10 patients with composite mandibular defects including mandibular body, mandibular angle, mandibular ramus and the soft tissue around them, 7 patients were recorded with the defects of mandibular condyles. During the follow-up period from 3 months to 24 months, primary wound healing was observed in all patients, except one patient with minor muscular necrosis. All patients were satisfied with their facial contour and mandibular shape, without tumor recurrence. Donor site problems important enough to be recorded in the notes were minimal.

CONCLUSIONSThe vascularized iliac crest graft with internal oblique muscle offers a useful solution for reconstruction of composite mandibular defect. There is sufficient height and depth of bone to maintain a facial contour and mandibular shape. It can be used as a routine surgical technique to reconstruct composite mandibular defect.

Abdominal Muscles ; transplantation ; Adolescent ; Adult ; Female ; Humans ; Ilium ; blood supply ; transplantation ; Male ; Mandibular Injuries ; surgery ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; Young Adult