Heat Stress Disorders ; etiology ; Humans ; Mining ; Occupational Diseases

AIM: To discuss the establishment of immediate diabetic keratopathy animal model of C57BL/6 mouse induced by ahigh-fat and high-glucose diet.
METHODS: Diabetes mellitus was induced by a high-fat and high-glucose diet in C57BL/6 mouse. 1% rose bengal was stained on the cornea to examine the integrality of the corneal epithelium at 2 ~ 12mo after completion of the model. Corneal epithelial wound healing was observed using a vivo epithelial debridement model which was dyed by sodium fluorescein. Corneal morphology histology was examined by pathological methods.
RESULTS: The high-fat and high-glucose diet C57BL/6 mouse in 2mo had showed general symptoms of diabetes: polydipsia, polyphagia, polyuria, weight loss etc. The model had a steady-state high glucose (≥18mmol/L), also the weight was lower compared with normal control mouse. 1% rose bengal corneal staining had dot coloring at 2mo after completion of the model, the stained area and extent were gradually increased with the extension of the duration of diabetes, almost all the cornea was stained at 12mo after completion of the
model. With the passage of time into a mold, the cornea epithelial healing time become longer: 2mo was about 40h;3mo was about 120h; 4, 6, 12mo was about 144h;the coloboma were gradually increased at 12mo after completion of the model, then the area was reduced gradually until complete healing, the time was 96~120h, showed repeating phenomenon.
CONCLUSION: The mouse were induced by high-fat and high-glucose diet can be used as animal models of diabetic keratopathy: the damage of epithelium for corneal and delay healing on epithelium and other symptoms.

OBJECTIVETo investigate the inhibitory effect of bone marrow mesenchymal stem cells (BMSCs) on E coliinduced prostatitis in rats.

METHODSBMSCs were isolated, cultured and amplified by the attached choice method. Fifty SD rats were randomized into five groups of equal number: normal control, acute bacterial prostatitis (ABP) , chronic bacterial prostatitis (CBP), ABP + BMSCs, and CBP + BMSCs, and the animals in the latter four groups were injected with E. coli into both sides of the prostate under ultrasound guidance for 1 - 14 days to induce ABP and for 4 - 12 weeks to induce CBP. The control rats were injected with the same amount of PBS. Two weeks after injection of BMSCs into the prostates, pathomorphological changes in the prostate were observed under the light microscope and the mRNA and protein levels of IL-1β and TNF-α determined by RT-PCR and ELISA, respectively, followed by statistical analysis with SPSS 18.0.

RESULTSHistopathological evaluation showed typical pathological inflammatory changes in the prostates of the rats in the ABP and CBP groups, including glandular structural changes, interstitial edema, inflammatory cell infiltration, and fibrous hyperplasia, which were all remarkably relieved after treated with BMSCs. The mRNA and protein levels of IL-β ([0.829 ± 0.121] and [271.75 ± 90.59] pg/ml) and TNF-α ([0.913 ± 0. 094] and [105.78 ± 19. 05] pg/ml) in the ABP and those of IL-1β ([0. 975 ± 0. 114] and [265. 31 ± 71. 34] pg/ml) and TNF-α ([0. 886 ± 0. 084] and [107. 45 ± 26. 11 ] pg/ml) in the CBP groups were significantly higher than those in the control rats ([0. 342 ± 0.087] and [45.76 17. 99] pg/ml, P <0. 05); ([0.247 ± 0.054] and ([19.42 ± 7. 75] pg/ml, P <0. 01) as well as than those in the ABP + BMSCs ([0. 433 ± 0. 072] and [51. 34 ± 22. 13] pg/ml, P < 0. 05 ) ; ( [0. 313 ± 0. 076] and [28. 38 ± 8. 78] pg/ml, P < 0. 01) and the CBP + BMSCs group ([0.396 ± 0.064] and [56.37 ± 21.22] pg/ml, P <0.05); ([0.417 ± 0.068] and [29.21 ± 10.22] pg/ml, P <0.01).

CONCLUSIONInjection of BMSCs can reduce E coli-induced prostatic inflammation reaction, which.may be associated with its reduction of inflammatory cell infiltration and the expressions of IL-1β and TNF-α in the prostate tissue.

Acute Disease ; Animals ; Bone Marrow Cells ; physiology ; Chronic Disease ; Escherichia coli Infections ; therapy ; Humans ; Interleukin-1beta ; genetics ; Male ; Mesenchymal Stromal Cells ; physiology ; Prostate ; metabolism ; Prostatitis ; metabolism ; microbiology ; therapy ; RNA, Messenger ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

A study was made on the pharmacokinetic regularity of effective components salvianolic acid B and ferulic acid in Salviae Miltiorrhizae Radix et Rhizoma (SMRR) and Chuanxiong Rhizoma(CR) in rats, so as to discuss the compatibility mechanism of Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma. Rats were randomly divided into three groups and intravenously injected with 50 mg x kg(-1) salvianolic acid B for the single SMRR extracts group, 0.5 mg x kg(-1) ferulic acid for the single CR extracts group and 50 mg x kg(-1) salvianolic acid B + 0.5 mg x kg(-1) ferulic acid for the SMRR and CR combination group. The blood samples were collected at different time points and purified by liquid-liquid extraction with ethyl acetate. With chloramphenicol as internal standard (IS), UPLC was adopted to determine concentrations of salvianolic acid B and ferulic acid. The pharmacokinetic parameters of salvianolic acid B and ferulic acid were calculated with WinNonlin 6.2 software and analyzed by SPSS 19.0 statistical software. The UPLC analysis method was adopted to determine salvianolic acid B and ferulic acid in rat plasma, including linear equation, stability, repeatability, precision and recovery. The established sample processing and analysis methods were stable and reliable, with significant differences in major pharmacokinetic parameters, e.g., area under the curve (AUC), mean residence time (MRT) and terminal half-life (t(1/2)). According to the experimental results, the combined application of SMRR and CR can significantly impact the pharmacokinetic process of their effective components in rats and promote the wide distribution, shorten the action time and prolong the in vivo action time of salvianolic acid B and increase the blood drug concentration and accelerate the clearance of ferulic acid in vivo.
Animals ; Apiaceae ; chemistry ; Benzofurans ; blood ; pharmacokinetics ; Coumaric Acids ; blood ; pharmacology ; Drug Interactions ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Rhizome ; chemistry ; Salvia miltiorrhiza ; chemistry

OBJECTIVEThe SSR information in the transcriptome of Erigeron breviscapus was analyzed in this study, in order to further develop new functional genes SSR markers laid a solid foundation.

METHODSSR loci were searched in all of 52,060 unigenes by using est_timmer. Perl program and SSR primers were designed by Primer3. Furthermore, 36 pairs of primers were randomly selected for the polymorphism analysis on 13 Erigeron breviscapus plants collected from different places.

RESULTA total of 3639 SSRs were found in the transcriptome of Erigeron breviscapus, distributed in 3260 unigenes with the distribution frequency of 6.99%. Di-nucleotide repeat was the main type, account for as much as 34.41% of all SSRs, followed by mono-nucleotide (31.41%) and tri-nucleotide repeat motif (28.08%). The di-nucleotide repeat motifs of AT/AT and AC/GT were the predominant repeat types (28.71%). The tri-nucleotide repeat motifs of AAT/AT was the predominant repeat types (7.94%). For validation the availability of those SSR primers, we randomly selected 36 pairs of primers for PCR amplification. Among them, 34 pair primers (94.44%) produced clear and reproductive bands, 19 pair primers showed polymorphism (52.78%), and 13 Erigeron breviscapus plants were divided into 2 groups.

CONCLUSIONThere are numerous SSRs in Erigeron breviscapus transcriptome with high frequency and various types, this will provide abundant candidate molecular markers for genetic diversity study and genetic map in this plant.

China ; DNA Primers ; genetics ; Erigeron ; classification ; genetics ; Genetic Variation ; Microsatellite Repeats ; Phylogeny ; Polymorphism, Genetic ; Transcriptome

Objective This study is to investigate the set-up accuracy of thermoplastic mask used for immobilization of nasopharyngeal carcinoma (NPC) patients being treated by Intensity Modulated Radia- tion Therapy (IMRT).Methods Nineteen patients with early stage nasopharyngeal carcinoma (T1- T2N0M0),treated by fractionated intensity-modulated radiation therapy,underwent repeated CTs during their 6-week treatment course.We evaluated their anatomic landmark coordinates in a total of 85 repeated CT data sets and respective x,y and z shifts relative to their position in the 19 treatment-planning reference CTs.Results The translation in x,y,and z-axes with their mean value derived from both positive and negative set-up errors was-0.84 mm(x-axis),+0.65 mm(y-axis) and +0.01 mm(z-axis).Mean target isocenter translation was (0.89?0.69) mm,(0.82?0.79) mm,(0.95?1.24) mm in x,y and z-direc- tions,respectively.The mean total magnitude vector and 95% CI of isocenter motion were 1.87 mm and 4.65 mm.The data measured over the 6-week fractionated course of treatment revealed a slight deterioration of isocenter accuracy.The 95% CI,considered by us to be a valuable parameter for characterizing the sys- tem,of 4.17 mm for measurement within the first 3 weeks increased to 5.12 mm in the last 3 weeks of treat- ment.Conclusions The sequential CT scanning is a pronounced valuable method of evaluating the quality of departmental specific patient positioning procedures.The thermoplastic mask is eyed as well suited tool for immobilization and repositioning of NPC patients receiving intensity-modulated radiation therapy.

AIMTo clone and analyse the coding cDNA sequence of alpha1, beta2 and gamma2 subunit of GABAA receptor in American king Pigeon.

METHODSWithdrew total RNA from the American king pigeon brain, reverse transcribing general primers to acquire a gene set cDNA. Designing specific primers of three subunit mRNA of the GABAA receptor, by RT-PCR respectively expanded the conservative gene of al subunit, beta2 subunit and gamma2 subunit of GABAA receptor, and carried on clone, plastid identification and the sequence measurese of three genes.

RESULTSThe experiment on sequence measures has succeeded that sequence analysis indicated that lengths of the conservative gene of alpha1 subunit, beta2 subunit and gamma2 subunit of GABAA receptor was respectively 899 bp, 597 bp and 563 bp, homology on reference sequence was respectively 94.99%, 94.64% and 96.28%.

CONCLUSIONHomology is high on the conservative gene of alpha1 subunit, beta2 subunit and gamma2 subunit of GABAA receptor of brain tissue of pigeon and chicken but there is a discriminating characteristic in different kinds of animals.

Animals ; Brain ; metabolism ; Cloning, Molecular ; Columbidae ; DNA, Complementary ; genetics ; RNA, Messenger ; genetics ; Receptors, GABA-A ; classification ; genetics ; Sequence Analysis, DNA

Background The pathological change of the anterior uveitis is the disruption of blood-aqueous barrier.Slit lamp examination appears to be limited for the evaluation of inflammatory response,and fluorescine angiography is an objective approach.However,there are few relative studies up to now in China.Objective Aim of this study was to observe the characteristics and assess the clinical applications of iris fluorescein angiography (IFA)in Chinese uveitis with brown iris.Methods Forty eyes of 40 normal subjects and 21 eyes of 13 patients with the anterior uveitis were collected in this study.IFA,slit-lamp examination and iris photograph were performed on the subjects.All individuals were informed consented at the initiation of this study.Results In normal eyes,fluorescence in iris vessels was blocked by the melanin pigment,but peripupillary weak fluorescent leakage was seen in the normal eyes with the age of ＞60 years old.The multiple patterns of fluorescence leakage were found in the patients suffered from uveitis of various etiologies although the negative slit-lamp finding,including the leakage of fluorescein around the pupillary margin and radial iris vessels in the eyes with mild diseases,transmitted fluorescence of regular iris vessels in the eyes with diffuse and local iris atrophy,and vascular tufts of the pupillary margin with coiled interwind tight clusters of thin vessels at the early phase in the eyes with dilated capillaries,microvascular anomalies and new vessel formation.The hyperfluorescence remained throughout the IFA duration.Conclusions IFA findings in uveitis vary depending on the topography,type and severity of inflammation.IFA has a good clinical applying value because of its objective assessment ability of the degree of the blood-aqueous barrier breakdown and iris neovascularization breakdown.It can exhibit the unvisible lesion under the slit-lamp and monitor the efficacy of medical theraphy in patients with active or quiescent uveitis.

Objective To explore the feasibility of constructing tissue engineered porcine corneal stroma with skin fibroblasts in vivo.Methods Skin fibroblasts were isolated from embryonic porcine,cultured and expanded in vitro.Cells were labeled with green fluorescence protein(GFP) gene by retro-viral infection.Cells at passage 3 were seeded on polyglycolic acid(PGA) non-woven fibers to form a cell-scaffold complex.The complexes were then implanted into porcines' corneal stroma after culturing in vitro for 1 week.Engineered stroma was observed continuously and harvested after 8 weeks for gross and histological evaluation.PGA with corneal stromal cells was served as control. Results The engineered tissue in the stroma gradually became transparent over a period of 8 weeks,showing no difference with the control group.Histologically,the engineered stromal lamellar was relatively regular and similar to the control.The implanted cells were confirmed by GFP expression under fluorescent microscope.By transmission electron microscopy examination, no significant difference in the diameter of collagen fiber was observed between the engineered stroma and normal stroma. Conclusion Tissue engineered corneal stroma may be formed with skin fibroblasts in porcine corneal microenvironment.

BACKGROUNDSuccessful lung transplantation has been limited by the scarcity of donors. Brain death (BD) donors are major source of lung transplantation. Whereas BD process induces acute lung injury and aggravates lung ischemia reperfusion injury. Carbon monoxide (CO) inhalation at 50-500 parts per million (ppm) can ameliorate lung injury in several models. We examined in rats whether CO inhalation in BD donor would show favorable effects on lung grafts.

METHODSRats were randomly divided into 4 groups. In sham group, donor rats received insertion of a balloon catheter into the cranial cavity, but the balloon was not inflated. In BD-only group, donor rats were ventilated with 40% oxygen after BD confirmation. In BD+CO250 and BD+CO500 groups, donor rats inhaled, after BD confirmation, 250 ppm or 500 ppm CO for 120 minutes prior to lung procurement, and orthotopic lung transplantation was performed. The rats were sacrificed 120 minutes after the lung transplantation by exsanguination, and their blood and lung graft samples were obtained. A total of 8 rats fulfilling the criteria were included in each group.

RESULTSThe inhalation decreased the severity of lung injury in grafts from BD donors checked by histological examination. CO pretreatment reversed the aggravation of PaO2/FiO2 in recipients from BD donors. The CO inhalation down-regulated pro-inflammatory cytokines (TNF-alpha, IL-6) along with the increase of anti-inflammatory cytokine (IL-10) in recipient serum, and inhibited the activity of myeloperoxidase in grafts tissue. The inhalation significantly decreased cell apoptosis in lung grafts, inhibiting mRNA and protein expression of intercellular adhesion molecule-1 (ICAM-1) and caspase-3 in lung grafts. Further, the inhalation activated phosphorylation of p38 expression and inhibited phosphorylation of anti-extracellular signal-regulated kinase (ERK) expression in lung grafts. The effects of CO at 500 ppm were greater than those at 250 ppm.

CONCLUSIONSCO exerts potent protective effects on lung grafts from BD donor, exhibiting anti-inflammatory and anti-apoptosis functions by modulating the mitogen-activated protein kinase (MAPK) signal transduction.

Administration, Inhalation ; Animals ; Apoptosis ; Brain Death ; Carbon Monoxide ; administration & dosage ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; Inflammation ; prevention & control ; Intercellular Adhesion Molecule-1 ; analysis ; genetics ; Lung Transplantation ; methods ; Male ; Phosphorylation ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Tissue Donors ; p38 Mitogen-Activated Protein Kinases ; metabolism