2.Changes in corneal nerve fiber length and density before and after eye opening in mouse and its significance
Cun, WANG ; Ting, FU ; Zhi-jie, LI ; Chao-yong, XIA
Chinese Journal of Experimental Ophthalmology 2012;30(7):581-585
Background Recently,there were many studies on corneal innervations during mammalian development.However,there were fewer studies on discussing corneal innervations before and after mouse eye openings.Objective The present study was to investigate the change in the regulation of corneal nerve fiber length and density before and after mouse eye openings to offer a basis for clinical research in human.Methods Thirty SPF C57BL/6 mice were divided into postnatal 1 day(P1 d),P7 d,P13 d(1 day before eye opening),P14 d(eye halfopened),P17 d(1 day after eye opening)and P23 d(7 day after eye opening)groups,with 5 mice and 10 eyes for each group.Entire corneal stretches were prepared and immunostaining with an anti-neuron-specific β-Ⅲ tubulin antibody was performed to label the corneal nerve fibers.Confocal microscopic pictures from the corneal dorsal-nasal region (DN),dorsal-temporal(DT),ventral-nasal region(VN)and ventral-temporal(VT)were taken using Delta Vision Core.From these pictures,the mouse corneal area,total length and density of nerve fibers in the 4 regions were calculated.The use of the animals complied with Statement of ARVO.Results Corneal areas of P1 d,P7d,P13 d,P14 d,P17 d and P23 d mice were(0.404±0.007),(1.362±0.154),(1.573±0.080),(1.603±0.046),(1.847±0.052),(2.445±0.798)mm2,respectively ; the total lengths of nerve fibers were(3.718±1.044),(19.065±3.350),(23.687±0.907),(27.309±2.477),(31.989±3.976),(41.214±1.573)mm,respectively ; the densities of nerve fibers were(9.592±1.138),(14.506±1.908),(15.088±1.241),(16.772±1.897),(16.821±2.102),(17.660±1.216)mm/mm2,respectively,all showing significant increases with age(F =22.906,P =0.000 ; F =0.424,P =0.000 ; F =2.375,P=0.000).A positive correlation of the increasing corneal areas and increasing lengths of nerve fibers was found(r=0.983,P<0.01).Nerve fiber densities in the four corneal regions significantly increased with age(DN region:F =0.159,P =0.000 ; DT region:F =2.1 72,P =0.001 ; VN region:F =1.998,P =0.000 ; VT region:F=2.352,P=0.000).From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0% without significant difference(t =0.589,P =0.572); and the corneal nerve fiber densities in the DT region,VN region and VT region decreased by 4.6%,5.5% and 0.1%,respectively,without significant difference from P14 d to P17 d(t=0.549,P=0.596;t=0.701,P=0.501 ;t=-0.100,P=0.919).Conclusions The development of nerve fibers in the whole cornea or the four corneal regions is influenced by eye opening in mouse to various extents.From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0% without significant difference.From P14 d to P17 d,the corneal nerve fiber densities in the DT region,VN region and VT region decrease by 4.6%,5.5% and 0.1%,respectively,without significant difference.Afterwards,the growth of nerve fibers increased in pace and the growth rate is recovered.
3.Using the SELDI Protein Chip System to Detect Changes in Protein Expression in Vero Cells after Infection
Zhi-jun, LIU ; Bin, WANG ; Zhi-yong, YAN ; Xu-xia, SONG ; Dong-meng, QIAN ; Zhi-qiang, BAI
Virologica Sinica 2007;22(1):68-73
Human herpes simplex virus 1 (HSV-1) causes facial,ocular,and encephalitic disease and is associated with latent infection and cancer.Here,we developed a means of studying the pathogenesis of HSV-1 infection at the protein level by using the SELDI Protein Chip to detect changes of protein expression in Vero cells cultured in vitro.After infection with HSV-1 and culture for 12,24 or 48 h,cells were harvested and lysed.IMAC3 arrays were applied to SELDI-TOF-MS to detect proteomic differences before and after infection.The chip detected a series of differentially expressed protein peaks.Interestingly,both peaks at 16 912 Da and 17 581 Da corresponded precisely with the molecular mass of ISG 15,which may participate in antiviral activity during the process of infection.Thus,the results we obtained can serve as a basis to study the pathogenesis of HSV-1 and the interaction between the virus and its host.In addition,they can help in the discovery of new therapeutic targets for treatment of HSV-1 infection.
4.Preparation of 131I-K237 and the experimental study on targeting therapy in nude mice bearing human lung cancer
Huo-qiang, WANG ; Jun-yong, XIA ; Zhi-yong, ZHANG ; Long, ZHAO ; Jin-xu, ZHENG
Chinese Journal of Nuclear Medicine 2010;30(6):390-394
Objective To establish the radiolabeling method for peptide K237 with 131I and investigate the biodistribution and therapeutic efficacy of 131I-K237 on nude mice bearing human lung cancer.Methods Iodogen method was used for labeling K237. The bioactivity of 131I-K237 was tested by human umbilical vein endothelial cell ( HUVEC ) proliferation inhibitory assay and the affinity of 131I-K237 was examined by competition binding studies. Twenty-five mice were divided into five groups randomly, including physiologic saline (group 1), K237 (40 μg) (group 2), 131I ( 11. 1 MBq) (group 3), 131I-K237 (K237 40 μg, 11. 1 MBq) intravenously ( group 4), and 131I-K237 ( K237 40 μg, 11.1 MBq) intratumorally (group 5). Injections were repeated at 15 d after the first injection. The tumor growth inhibition rate was calculated. Student's t-test and analysis of variance (ANOVA) were used for testing significant differences of data. Results The inhibition rate of HUVEC proliferation had no significant difference between radiolabeled K237 and unlabeled K237 ( (73.69 ± 5.36) % vs ( 62.68 ± 3.83 ) %, t = 1.67, P > 0.05 ). The growth of transplanted lung cancer was inhibited by 75. 01 % in group 4, 78.99% in group 5, 31.15% in group 2 and 12.61% in group 3, respectively. The average tumor volume of groups 4 and 5 were significantly smaller than that of groups 1,2, and 3 ( F = 15. 233 and 13.611, respectively, P <0. 01 ). Conclusion 131I-K237 can be readily radiolabeled and it can effectively inhibit the growth of tumor in nude mice bearing human lung cancer.
5.Progress of studies on genetics of childhood acute leukemia.
Journal of Experimental Hematology 2013;21(3):791-795
This study on determination of leukemia-specific chromosomal abnormalities and their relationship with prognosis of childhood acute leukemia (AL) had an important significance for childhood acute leukemia. In recent years, the efficacy of treatment of childhood AL has been greatly improved, but relapse is still a main factor affecting prognosis. Treatment based on the risk stratification by cytogenetic abnormalities can improve the prognosis and survival rate. In the past 3 decades, the genetic techniques have developed rapidly and many new genetic abnormalities have been found. This review highlights the main chromosomal and genomic abnormalities of 3 common childhood AL, including B-cell precursor acute lymphoblastic leukemia (BCP-ALL), T-cell acute lymphoblastic leukemia (T-ALL) and acute myeloid leukemia (AML).
Acute Disease
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Child
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Humans
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Leukemia
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genetics
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Leukemia, Myeloid, Acute
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genetics
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Precursor B-Cell Lymphoblastic Leukemia-Lymphoma
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genetics
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Precursor T-Cell Lymphoblastic Leukemia-Lymphoma
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genetics
6.Trabeculectomy punch combined with a novel RS tube for the treatment of punctal stenosis
Yong-Zhe, TANG ; Hong-Ling, LYU ; Hai-Zhi, MA ; Xiao-Xia, LIU ; Kang-Fu, LIANG
International Eye Science 2017;17(7):1365-1367
AIM: To investigate the feasibility and clinical effect of punctoplasty by using trabeculectomy punch combined with a novel RS tube for the treatment of punctal stenosis.METHODS: Totally 39 patients (39 eyes) with punctual stenosis were selected from October 2013 to October 2015 in the Second People`s Hospital of Foshan.All patients underwent punctoplasty by using trabeculectomy punch combined with a novel RS tube.These tubes were removed at 3mo after operation.A follow-up of 6mo was taken for final analysis.The fluorescein dye disappearance test score was recorded before the operation and at 1,3 and 6mo after the extubation.The curative effect of the operation at 6mo after the extubation was assess.RESULTS: Fluorescein dye disappearance test: the scores at 1,3 and 6mo after the extubation all decreased compared with the preoperative ones.The difference was statistically significant(P<0.05).At the last following up, 35 eyes (90%) were cured completely, 4 eyes (10%) were improved significantly, no patients recurred.Effective rate was 100%.No serious intraoperative and postoperative complications happened.CONCLUSION: Punctoplasty by using trabeculectomy punch combined with novel RS tubes is a safe and effective method for the punctul stenosis, which is easy to perform, with high success rate.
7.Hepatoprotective Effects of Silybin-Phosphatidylcholine Complex in Septic Rats
xing-xia, LIU ; pei-yong, QIU ; jian-zhi, GAO ; zheng-yue, CHEN
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To investigate the hepatoprotective effect of silybin-phosphatidylcholine complex(SPC) in sepsis rats.Me-(thods) Fifty Wistar immature rats were randomly divided into 3 groups:normal control(10 cases),septic group(30 cases) and interfe-(rence) group(10 cases).In septic group and interference group,rats were treated by cecal ligation and puncture(CLP).Meanwhile,the interference group was given SPC before 2 hours of CLP and after 2 hours of CLP.Blood of rats was taken from all groups to determine the levels of serum tumor necrosis factor-?(TNF-?),interleukin-1(IL-1),alanine aminotransferase(ALT) and aspartate aminotransferase(AST) at varying intervals.Results The levels of serum TNF-?,IL-1,ALT and AST after CLP were significantly elevated in septic group compared with the normal control group(P
8.Nutrition Factors Influence the Production of Insecticidal Crystal Proteins Cry1 and Cry2 from Bacillus thuringiensis 4.0718
Fei LIU ; Li-Qiu XIA ; Xue-Zhi DING ; Yong YI ; Xiang-Tao MO ; Wei WEI ;
Microbiology 2008;0(08):-
In order to increase the production of insecticidal crystal proteins Cry1 and Cry2, firstly, Plack-ett-Burman design was applied to evaluate the effectiveness of the related nutrition factors; it was found that the soybean powder and MnSO4?H2O were significant factors for Cry1 production, but the yield of Cry2 wasn’t effected remarkably in such medium. Then the steepest ascent experiment was adopted to approach the optimal region of the medium composition. Lastly, the optimal concentration of the soybean powder and MnSO4?H2O was 11.5 and 0.02 g/L, obtained by response surface methodology (RSM). The final yields of Cry1 and Cry2 was 0.32 mg/mL and 0.11 mg/mL, increasing twice more than that in the medium optimized before. The median lethal concentration (LC50) of optimal medium was 1.09 ?L/mL. The toxicity to Heli-coverpa armigera was significantly enhanced than the old one.
9.Studies on Shedding Virus and the Formation of Aerosol of H_9N_2 Subtype AIV
Yong-Zhi CAO ; Wei-Ming MA ; Xiao-Xia LI ; Tong-Jie CHAI ; Hai-Yan ZHANG ;
Microbiology 2008;0(10):-
In order to study the regularity of shedding virus from infected SPF chickens and the formation of aerosol of H9N2 subtype AIV, SPF chickens were bred in a positive and negative pressure isolator. Aerosol samples were collected by AGI-30 (All Glass Impinger-30) extractor, and simultaneously trachea and cloaca samples were collected by tracheal swabs and cloacal swabs in different periods after challenged with vi- ruses. The above-mentioned samples were detected by HI, Dot-ELISA and RT-PCR methods. The results in- dicated that aerosols were isolated from the 4 days to the 43 days after inoculation. It was proved that H9N2 subtype AIV could copy themselves in respiratory passage and cloaca, and then could formation of aerosols. AIV H9N2 subtype could be isolated from cloacal and tracheal swabs 3 days after inoculation and lasted for 45 days, viruses were detected from all infected SPF chickens on 7 days.
10.Molecular typing of methieillin-resistant Staphylococcus aureus isolated from hospitalized patients in Qingdao
Fu-Guo YANG ; Zhi-Yong YAN ; Chun-Xia BI
Chinese Journal of Epidemiology 2008;29(12):1230-1234
Objective To explore the molecular types of methicillin-resistant Staphylococcus aureus (MRSA) strains present in major hospitals in Qingdao area, using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) methods, trying to find out the epidemiological characteristics of these MRSA isolates. Correlation of the PFGE types with microbiological phenotypes and clinical data was also studied. Methods 360 isolates of MRSA were procured during 2003 to 2007 from major hospitals in Qingdao. PFGE technology was applied to comparatively analyze the chromosomal DNA digested with endonuclease Sma Ⅰ . Comparison of DNA fragments patterns from each MRSA strain and cluster analysis were performed with the Bionumericus version ' 2.0' software. A dendogram was generated using PFGE macrorestriction fragments on gel images. Data was used to predict the possibility of each PFGE type via SPSS software version 11.0, using the variables as predictors including groups on patient's age, gender, source and the site where MRSA was isolated. Antibiotic sensitivity patterns of these MRSA isolates were determined by K-B tests, and a correlation between these patterns and PFGE types was investigated. Housekeeping genes were amplified with PCR and sequenced in representative strains of variant PFGE types to identify their allelic profile. Results 5 types of PFGE patterns (M0-M4) were identified with MI being the predominant and M2 next to it which was significantly correlated to the isolates from wounds. M3 type strains were mainly isolated from ICU wards and there were a few cases complied with M4 type with no correlated variant factors found in this study. A unique pattern of MRSA isolates with its M0 distinct from other types had not been reported. No significant association was found between PFGE individual types,gender or age groups. M1 and M2 types were the major proportional PFGE patterns among different hospitals. No vancomycin-resistant isolates were detected among 360 MRSA strains. No significant association was found between individual antibiotic resistance and specific PFGE types. Data from MLST analysis showed that the aUelic profiles of M1 and M3 type strain had the same ST239 linage which was commonly present in China. For M2 and M4 representative strains, the allelic profiles were ST5 and ST240, respectively. ST45 and ST398 were corresponding to two PFGE patterns clustered as M0 type. Conclusion Nosocomial infection due to MRSA was evenly distributed among different age groups and no gender bias was observed. The PFGE types of MRSA strains isolated in major hospitals in Qingdao were highly correlated with the sources of isolates and ST239 isolate seemed the prevalent and widespread one. Strategies should be designed to further monitor and prevent or minimize the spread of ST5 MRSA isolates and the like, in Qingdao area.