Aged ; Altitude ; Endothelin-1 ; blood ; Female ; Humans ; Hypertension, Pulmonary ; Male ; Middle Aged ; Pulmonary Heart Disease ; physiopathology ; Ventricular Function, Right

Background In the past few decades,the balance of Th1/Th2 is often used to explain the immune mechanisms of fungal infection and fungal disease.More recently,a novel subset of CD4+ effector Th cells has been found to participate in anti-fungal infection response.However,whether Th17 is involved in the immune response in fungal keratitis is unclear up to now.Objective Present study was to investigate the expression change of Th17 type cytokine and its specific transcription factor,retinoid-related orphan nuclear receptor gamma t (RORγt),in the cornea of Fusarium solani keratitis.Methods Ninety-six clean BALB/c mice were divided into Fusarium solani keratitis model group and control group by randomized digital table.Fusarium solani keratitis models were established by epikeratophakia-assisted corneal epithelial erasion and interlayerly injection of 5 μl (1 × 106 CFU/ml) Fusarium solani solution in the right eyes,and the equal volume of PBS was injected in the same way in the control group.10％ KOH wet film was used to examine the fungal hyphea and funga strain was identified by inoculation.The corneas were examined under the slit lamp microscope 1 day,3,5,7 days after modeling and the inflammatory response was scored based on the criteria of Wu and Hu.The histopathological examination of corneas was performed in the time points above.Real time fluorescence quantitative PCR was used to detect the expression levels of interleukin-17 (IL-17) mRNA and RORγt mRNA in the corneas.The expression of IL-17 protein in the corneas was detected by ELISA.The use and raise of the mice followed the Statement of Association for Research in Vision and Ophthalmology.Results The inflammatory scores were 3.2±0.8,6.6± 1.1,9.4± 1.1 and 6.8±0.8 in 1 day,3,5,7 days after modeling,showing a significant difference among them (F =89.786,P =0.010).The inflammatory scores were higher in the third and seventh day than that in the first day (P＜0.05),but they were significantly lower than that in the fifth day (P＜0.05).The infiltration of inflammatory cells showed a coincident tendency with the score.The expressing levels of IL-17 mRNA (2-ΔΔCt) in the corneas were 4.12±0.73,20.72±1.81 and 14.16±1.88 in 3,5,7 days after modeling,with statistically significant differences in comparison with those in the control group (P＜0.01),and the expression level was significantly higher in the fifth day than those in the first,third and seventh day in the model group(P＜0.01).The expression levels of IL-17 protein (ng/g) were significantly increased 1 day,3,5,7 days in the model group compared with the control group (P＜0.01).A similar change was found in the expression of RORγt mRNA to that of IL-17 mRNA.Conclusions Expressions of IL-17 and its transcription factor RORγt upregulate in the fungal keratitis and has an association with inflammatory degree,which suggests that Th17 subset may play an important role in the immune responses of fungal keratitis.

Objective To analyze the imaging characteristics of thoracic LDRD(Low-dose directly Digital Radiographic Device) and its artifacts.Methods 188 patients were performed with LDRD and common thoracic X-ray film respectively in our hospital during two weeks.Results Among the 188 cases,1.60%(3/188) showed thoracic motion artifacts.46.8%(88/188) appeared as tentorial prominence along left heart edge and 2.6%(5/188) along the right one.1 artifact was in aorta-pulmonary artery window(0.53%).Conclusion(1)Less than 0.5 should be taken as reference in podoid enlargement diagnosis.(2)Pseudomorph from heart motion may result from cardio-phase,cardiac contraction,heart rate,arrhythmia,local abnormal pulse of left heart edge,different enlarged velocity of cardiac cavity during heart beat,etc.(3)The motion artifacts in thoracic LDRD has no important influence in clinical diagnosis and therapy.

Background Oxidative damage plays an important role in pathogenesis of age-related macular degeneration( AMD ),and its mechanism is the destroy of blood-retinal barrier.Müller cells is a primary component to stabilize the inner barrier of the blood-retina.Researches showed that epidermal growth factor(EGF) can promote the proliferation and migration of animal Müller cells,but less study was found in the effect of EGF on human Müller cells. Objective The present study was to investigate the effects of EGF on the proliferation and migration of human Müller cells and its molecular mechanism. Methods Human Müller cell line MIO-M1 cells were cultured and incubated,and cultured cells were identified using glial fibrillory acidic protein (GFAP),factor Ⅷ,α-smooth muscle actin( α-SMA ),keratin and S-100.Different concentrations of EGF( 0,1,10,30,100 mg/L)was added in freeserum DMEM,and the positive rate of the cells was calculated using 5-bromo-2-deoxyuridine(BrdU) method.The cells were divided into EGF group,H2 O2 group,EGF + H2 O2 group,glucose oxidase ( GO ) group,GO + EGF group,EGF + LY294002+H2O2 group according to the different intervention,and the effects of LY294002 on the proliferation of Müller cells (A590 )were detected by colorimetric assay for cellular growth and survival( MTT assay).The scratch test of Müller cells was used to assess the influence of EGF(0,1,10,30,100 mg/L)on H2 O2-induced damage of human Müller cell.Western blot was used to detect the cell proliferation under the protection of EGF on co-cultured cells using LY294002 and H2O2 and the activation of Akt signal pathways. Results The proliferative rates of the cells were 28.0％,32.9％,39.0％ in 10,30,100 mg/L EGF groups respectively and obviously higher than those in 0,1 mg/L EGF groups (24.5 ％,26.2 ％ ).Under the H2O2 culture,GO culture,respectively,the A570 value of the Müller cell in high concentrations of EGF groups was significantly increased in comparison with lower concentrations EGF groups with the statistical significance among the groups( F=23.582,P=0.000).Compared with EGF+H2O2 group,the A570value of the Müller cells was lowed in EGF+LY294002+H2O2 group.The maximum migration rate of Müller cells was found in 10 mg/L EGF group.Western blot revealed that the presence of H2O2 reinforced the expression of Akt in Müller cells,however,pretreatment with 100 mg/L EGF antagonized the harmful effect of H2O2 on Müller cells.Meanwhile,pretreatment with EGF and LY294002 reduced the expression of Akt in Müller cells. Conclusions EGF can induce the proliferation and migration of human Müller cells with the strongest effect in 10 mg/L.100 mg/L exogenous EGF has a stronger protection to the Müiller cells against H2O2-induced cell damage by activating the PI3KAkt cell survival pathway.

Objective To provide a practicable method for the complete display and localization of the posterolateral structures (PLS) of the normal knee through MRI study. Methods 30 tibial bone specimens were observed to establish the bony landmark for localizing the knee. In 50 cadaver knees, the angles between lateral tibial plateau and the long axis of the individual structure of PLS were measured. Then the scan methods of the oblique MR images were determined based on above results. The routine and oblique scans of T 1WI were performed in 40 normal knees. The display effect and appearance of the PLS were observed on MRI. Results The lateral tibial plateau was a stable bony landmark for measuring and localizing of the knee. In the 40 normal knees, The fibular collateral ligament could be intactly displayed on 70? posterior coronal oblique images in 34 cases (85%). The popliteus could be better seen on either 45? medial sagittal oblique in 34 cases (85%) or 60? posterior coronal oblique planes in 36 cases (90%). The popliteofibular ligament could be intactly appreciated on both 60? posterior coronal oblique in 32 cases (80%) and 70? lateral sagittal oblique images in 34 cases (85%). Although the arcuate ligament and the fabellofibular ligament could occasionally be seen on routine and oblique images, but the display rate was lower. Conclusion The oblique MR imaging can intactly display the main structures of PLS, and can be useful in diagnosing the injuries in those structures.

Objective To investigate the effect of escharectomy on rats' pulmonary NF-?B activation and the expression of pulmonary proinflammatory cytokines in early stage of burn injury.Method Wistar rats were randomly divided into three groups:group A(control group),group B(postburn without escharectomy),group C(escharectomy at early stage of burn injury).Thermal-injuried rats underwent 35% TBSA full-thickness burns. Activation of pulmonary NF-?B at 12 hours and 24 hours postburn was tested by electrophoretic mobility shift assay (EMSA),and at the same time expressions of pulmonary TNF-?mRNA were measured by reverse transcription polymerase chain reaction(RT-PCR)and release of pulmonary TNF-?were assayed by enzyme-linked immunosorbent assay(ELISA).Results Compared with control group,activity of pulmonary NF-?B in group B was markedly increased,reached(19.56?1.36)?10~4 A at 12 hours and(15.23?1.94)?10~4 A at 24 hours,which was higher than that in group A[(4.36?0.38)?10~4 A,P

Objective To isolate and identify the adult neural stem cells from the parenchyma of spinal cord in adult mouse.Methods The parenchymal spinal cord from adult mouse was dissected and dissociated by mechanical trituration.The tissue suspension was cultured in serum-free DMEM/F12 medium supplemented with EGF and B27.The cell colonies generated from a single cell were screened by limited dilution and incubated with BrdU.The cell colonies were transferred into medium with serum to induce differentiation.The cells were identified with antibodies to Nestin,BrdU,MAP2 and GFAP by immunofluorescence staining.Results The cells were cultured for seven days to generate proliferative neurospheres.The majority of cells in these neurospheres expressed Nestin and were differentiated into MAP2-positive cells and GFAP-positive cells in medium containing with fetal bovine serum.Conclusion A significant number of neural stem cells are present in the parenchymal adult mouse spinal cord and can proliferate and also give rise to neurons and glia in vitro.

Objective To study the effects of efflux pump inhibitors(CCCP and PAβN)on carbapenems in Pseudomonas aernginosa(P.aeruginosa)clinical isolates and investigate the association between the resistance to imipenem or meropenem and expression levels of efflux pumps of P.aeruginosa.Methods MICs of imipenem or meropenem combined with efflux pump inhibitors including carbonyl cyanide m-chlorophenylhydrazone(CCCP,107 strains)and Phe-Arg-β-naphthylamide(PAβN,71 strains)against imipenem-resistant strains were determined by agar dilution method,and changes of MICs were observed.For 32 strains with different resistant phenotypes to imipenem and meropenem,the mRNA expression levels of three efflux pump genes(mexA,mexD and mexF)were quantified by real time fluorescent quantitative PCR.Results The resistance rate of imipenem and meropenem didn't prove any significant difference in the presence of efflux pump inhibitors.The X2 value of imipenem combined with CCCP and PAβN were 0.338 and 0.086,respectively(P＞0.05),while that of meropenem combined with CCCP and PAβN were 1.065 and 1.458(P＞0.05).No significant in MICs of carbapenems were seen in over half of P. aeruginesa isolates. MICs of carbapenems was significantly downregulated for 4-fold or above in eight isolates. Overexpression of efflux pumps genes were present in 24 of 27 carbapenem-resistant isolates(88. 9% ). Efflux pumps genes including MexAB-OprM, MexCD-OprJ and MexEF-OprN were all overexpressed in 13 isolates,constituting 54. 2% of all carbapenem-resistant isolates. There were 3 isolates in which beth MexAB-OprM and MexCD-OprJ showed overexpression,constituting 12. 5%. Also,MexAB-OprM and MexEF-OprN overexpressed in 3 isolates. There were 2 isolates (8.3%) showing MexEF-OprN overexpression and MexAB-OprM alone. MexCD-OprJ didn't showed overexpression alone. Furthermore,the expression levels of efflux pumps genes mexA,mexD and mexF in isolates susceptible to both in imipenem and meropenem were 0. 48±0. 48,0. 48±0. 53 and 0. 30±0. 41,respectively,which were much lower than that in carbapenem-resistant ones (P＜0. 05 ). MexA gene was expressed at a higher level in meropenemresistant isolates than meropenem-susceptible ones (P＜0. 05 ). Conclusions When the concentration of CCCP and PAβN were 5 μg/ml and 20 μg/ml respectively,the efforts on the carhapenems resistance of P.aeruginosa were small Overexpression of MexAB-OprM might play an important role in meropenemresistance in P. aerugines. Overexpression of MexCD-OprJ and MexEF-OprN was associated with imipenemresistance. However,the relationship between them and meropenem-resistance need to be explored in the future.

Objective To assess the endemic trend of Kaschin-Beck disease in Tibet and to provide scientific basis for prevention and etiology study of the disease. Methods A questionnaire designed by us was administered to 905 participants who were from Lhundrop county, Medro Gongkar county of Lhasa municipality and Sangri county of Lhoka region in July to November, 2007. The Kashin-Beck disease diagnostic criteria(GB 16003-1995) was used for clinical diagnosis, and children 5 to 14 years old were taken right wrist X-ray film for diagnosis.Results One hundred and forty-four genealogies were recruited in this study. The interview and clinical examination were done to 905 persons, 208 persons were detected with Kaschin-Beck disease, and the detectable rate was 22.98%(208/905). The numbers of patients with degrees Ⅰ , Ⅱ and Ⅲ of Kaschin-Beck disease were 148, 43 and 17, respectively, with proportion of 71.15%(148/208), 20.67%(43/208) and 8.17%(17/208) out of all patients, respectively. The detectable rates of Kaschin-Beck disease were 29.73% (102/343) and 18.86%(106/562), respectively in Lhasa and Lhoka district, and the difference between this two districts was statistically significant(x2= 15.257, P＜ 0.01) . A total of 368 males and 537 females were recruited in this study, the detectable rates of male and female with Kaschin-Beck disease were 19.29% (71/368) and 25.51% (137/537), respectively,and the difference between male and female was statistically significant (x2 = 5.372, P ＜ 0.01) . In this study most patient were between 31 to 70 years old, the patients with degrees Ⅱ or Ⅲ of Kaschin-Beck disease were mostly above 40 years old. There were only 5 patients who were less and equal 20 years old in chinical diagnosis. The Xray positive detectable rate of children between 5 to 14 years old was 6.85% (10/146). Conclusions The condition of Kashin-Beck disease area is relatively stable in these two regions in recent years, and shows a downward trend. However, there are still positive child cases diagnosed by X-ray, which should arouse the attention of the relevant departments to further strengthen the implementation of control measures.

OBJECTIVETo explore the changes in the mRNA expression of adiponectin (Adp), adiponectin receptors(AdpR), and leptin in different adipose tissues of Wannanhua pigs at different stages of development, and their sexual dimorphism.

METHODSFive Wannanhua boars and five Wannanhua gilts were sampled at birth, 30, 45, 90, and 180 days of age respectively. The delta delta Ct relative quantification real-time PCR was used to detect the transcription levels of Adp, AdpR1, AdpR2, and leptin mRNAs in subcutaneous (SC) and perirenal (PR) adipose tissues, and beta-actin were used as internal standards.

RESULTSThe expression level of Adp, AdpR1, AdpR2, and leptin mRNA in SC and PR adipose tissue were changed with age significantly (P < 0.01). In general, Adp mRNA expression in SC adipose tissue was significantly lower than that in PR adipose tissue (P < 0.05), while AdpR1, AdpR2, and leptin mRNA expression in SC adipose tissue were significantly higher than those in PR adipose tissue (P < 0.05 or P < 0.01). Although the sexual dimorphism were found in apart genes or apart days of age, Adp, AdpR1, AdpR2, and leptin mRNA expression both in SC adipose tissue and PR adipose tissue had no significant differences between Wannanhua gilts and boars in general. Significant positive correlation was found between Adp and AdpR1, AdpR2 (P < 0.05 or P < 0.01), and significant negative correlation was found between Adp and leptin (P < 0.05) in SC adipose tissue and PR adipose tissue respectively (P < 0.05).

CONCLUSIONThe expression of Adp, AdpR1, AdpR2, and leptin mRNA in adipose tissue of Wannanhua pigs followed specific developmental patterns and tissue specificity. Adp correlated with its receptors.

Actins ; metabolism ; Adiponectin ; metabolism ; Adipose Tissue ; growth & development ; metabolism ; Animals ; Female ; Leptin ; metabolism ; Male ; RNA, Messenger ; genetics ; Receptors, Adiponectin ; metabolism ; Swine