Objective To monitor the function of infection on myelination in white matter damage,neonatal Wistar rats of postnatal day 2 (P2) and postnatal day 7 (P7) were injected intraperitoneally with the same doses of lipopolysaccharides (LPS),and 2',3 '-cyclic nucleotide phosphodiesterase (CNPase) and myelin basic protein (MBP) were labeled in immature oligodendrocytes and mature oligodendrocytes.To investigate the function of tumor necrosis factor(TNF)-α according to test the change of TNF-α expression in the brain.Methods Ninty-six neonatal Wistar rats were randomly divided into four groups (each group 24 rats):group A:LPS (5.0 mg/kg) was injected intraperitoneally on P2 ; group B:LPS (5.0 mg/kg) was injected intraperitoneally on P7 ;group C1 and C2 were control groups in which equal amount of normal saline was injected intraperitoneally on P2 or P7.The expression of CNPase at 24 h after injection and MBP at P14 in brain tissue of each group were measured by immunohistochemistry and express of TNF-α mRNA at 4 h after the injection was measured by RT-PCR.Results Punctate hemorrhage in the corpus callosum,external capsule and intraventricular hemorrhage were seen in group A.Periventricular leukomalacia appeared in the corpus callosum and glial cells hyperplasia could be seen periventricular in P14 rat brains,but not found in the group B and any of the saline-injected rat brains.Compared with group C1 and C2 respectively,CNPase-positive cells showed obvious decrease in the area of white matter in periventricular in group A(106.93 ± 2.62 vs 113.67 ± 2.69,P ＜ 0.01) and group B (96.37 ± 1.82 vs 101.65 ± 2.01,P ＜ 0.01).Following LPS treatment in group A,the protein expression of MBP in neonatal brain decreased evidently compared with group C1 at P14 (128.21 ± 2.99 v s 134.81 ± 2.98,P ＜ 0.01),while no significant change was found between group B and group C2(134.77 ±3.68 vs 134.81 ±2.98,P ＞0.05).After 4h of the LPS treatment,the level of TNF-α mRNA was greatly increased in group A,it was significantly higher than that in group B (1.79 ± 0.04 vs 1.18 ± 0.04,P ＜ 0.01).Conclusion Intraperitoneal injection of LPS to the development neonatal rats can lead to dysmyelination and white matter damage.The expression of TNF-oα mRNA increased significantly in these immature neonatal rats,while only myelination delay occurred in those of mature neonatal rats without dysmyelination.

Animals ; Colonoscopy ; methods ; Colorectal Neoplasms ; pathology ; Disease Models, Animal ; Heterogeneous-Nuclear Ribonucleoproteins ; metabolism ; Precancerous Conditions ; pathology ; Rodentia

AIM:To observe the clinical effect of femtosecond laser assisted penetrating keratoplasty.
METHODS: Twenty-four cases ( 24 eyes ) with corneal lesions were performed with femtosecond laser assisted penetrating keratoplasty. Preoperative and postoperative endothelial cell density and visual quality were compared.RESULTS: One week after operation, corneal grafts were clear in 21 eyes (87. 5%), mild cloudy in 3 eyes (12.5%);visual acuity ≥0. 5 in 18 eyes (75. 0%), 0. 2 ~0.4 in 6 eyes ( 25. 0%). After 3mo the mean corneal astigmatism was 2. 16±0. 21D ( range 2. 25 ~ 3. 09D). Compared to conventional penetrating keratoplasty which mean corneal astigmatism was average 3. 67±0. 38D after operation, there was significant difference between two groups ( P< 0. 05 ). There were significant differences between preoperative and postoperative visual acuity and astigmatism (both P<0. 05).
CONCLUSION: Femtosecond laser assisted penetrating corneal transplantation operation can improve patient's visual quality. And compared to traditional penetrating keratoplasty astigmatism decreased significantly, incision can be made in individual shape more precisely and neatly.

?AlM: To analyze and study the changes of intraocular pressure ( lOP) , visual field and P-ERG on patients with big cup/disk (C/D) in 24h.
?METHODS: A total of 120 cases ( 240 eyes ) diagnosed with big C/D (C/D>0. 3) were divided into group A (C/D<0. 6, 67 cases, 114 eyes ) and group B ( C/D≥0. 6, 73 cases, 126 eyes). Forty cases (80 eyes) with small C/D (C/D≤0. 3) were chosen as control group. All cases underwent 24h lOP examination, vision examination of 30-2 SlTA - standard static threshold and pattern electroretinogram ( P-ERG) examination. The differences between the examination indexes of the three groups were analyzed.
?RESULTS:There was no significant statistical difference in the 24h average lOP (P>0. 05) among the three groups, while the amplitudes had significant statistical differences (P<0. 05). Compared with the control group, the lOP amplitudes of group B were obviously higher, and the difference was of statistical significance (P<0. 05). ln terms of static threshold visual fields, the mean deviations (MD) and pattern standard deviations (PSD) of the three groups had significant statistical differences ( P < 0. 05 ). Comparison between every two groups:compared with the control group, the MD and the PSD were significantly increased in group B, and the difference was of statistical significance (P<0. 05); while there was no significant statistical difference in group A (P>0. 05);Compared with group A, the MD and the PSD were significantly higher in group B, and the difference was of statistical significance (P<0. 05). ln terms of P-ERG examination, there was no significant statistical difference in P50’s latent periods among the three groups (P>0. 05), but there were significant statistical differences in amplitudes (P<0. 05). Comparison between every two groups: compared with the control group, the P50’s amplitude was significantly decreased in group B, and the difference was of statistical significance (P<0. 05), while there was no significant statistical difference in group A (P>0. 05); Compared with group A, the P50’s amplitude of group B was significantly lower, and the difference was of statistical significance (P<0. 05).
?CONCLUSlON: When the patients’ C/D is no less than 0. 6, there are obvious changes of 24h lOP, static threshold visual field and P-ERG’s P50 wave. For patients with big C/D, a single lOP examination is far from sufficient, so an in-depth visual function examination should be performed. Meanwhile, it’s feasible to regard C/D 0. 6 as a screening criterion for suspected glaucoma.

Aim To establish a model of the vascular endothelial cell (VEC) injury in SDrats.Methods SD rats were randomly divided into the control and the modelgroups. The model rats were injected with adrenaline diluted to 2. 5 times 0. 05 mg?100 g-1 (tid) for 5 d continously. From the 4th d, they were irritated for 5 min in the0℃ cold-water in the middle between adrenaline injections.The control rats weregiven 0. 9% NS as above. At 6th d, blood samples were taken from carotid arteries ofthe rats and the CEC counts, t - PA、PAI activities, 6-keto-PGF1? concentrations andthe platelet aggregation rate(max) were detected respectively. Results In the modelgroup, as compared with those in the control group, t - PA activity and 6-keto-PGF1?concentration decreased significantly(P

This study is to investigate the effect and mechanism of puerarin on DNA damage of HaCaT cells induced by UVB. Puerarin pre-treated cells were irradiated with UVB at 30 mJ x cm(-2). Twenty four hours after irradiation, DNA damage was detected by comet assay, ceramide was measured by thin layer chromatography and gas chromatography, intracellular free calcium ion was analyzed by flow cytometry, the phosphorylation level of p38 protein was examined by Western blotting method. Levels of DNA damage, ceramide, free calcium ion and p-p38 protein were elevated in UVB model cells. Contrary to the model group, all indicators above were reduced in all groups pre-treated by puerarin. Puerarin restrains the ceramide accumulation to block downstream p38 MAPK pathway and calcium ion rising, therefore reduces DNA damage in HaCaT cells induced by UVB.
Calcium ; metabolism ; Cell Line ; Ceramides ; metabolism ; DNA Damage ; drug effects ; radiation effects ; Down-Regulation ; Humans ; Isoflavones ; pharmacology ; Keratinocytes ; cytology ; metabolism ; Phosphorylation ; Signal Transduction ; drug effects ; Ultraviolet Rays ; adverse effects ; p38 Mitogen-Activated Protein Kinases ; metabolism

A method of LC-QTOF/MS combining with chemical synthesis has been used to determine the structures of three novel bile acids from bear bile powder. Reference substances of tauroursodeoxycholic acid and taurochenodeoxycholic acid were oxidized by pyridinium chlorochromate. The products were analyzed by LC-QTOF/MS. Total 4 products including 3 isomers were predicted and identified according to the PCC oxidation theory and LC-QTOF/MS results. Bear bile powder samples were dissolved by methanol and analyzed by LC-QTOF/MS. Three unknown peaks were found and identified as 2-[[(3beta, 5beta)-3-hydroxy-7, 24-dioxocholan-24-yl]amino]-ethanesulfonic acid, 2-[[(5beta)-3, 7, 24-trioxocholan-24-yl]amino]-ethanesulfonic acid and 2-[[(5beta, 7beta)-7-hydroxy-3, 24-dioxocholan-24-yl]amino]-ethanesulfonic acid, separately, by matching their results with that of oxidation products above.
Animals ; Bile ; chemistry ; Bile Acids and Salts ; analysis ; chemistry ; Chromatography, Liquid ; methods ; Isomerism ; Molecular Structure ; Oxidation-Reduction ; Powders ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Taurochenodeoxycholic Acid ; chemistry ; Ursidae

Objective To study the exact function of human gene 5 transactivated by pre-S1 protein of hepatitis B virus(PS1TP5)by investigating the gene expression of PS1TP5 in yeast cells. Methods Reverse transcription-polymerase chain reaction(RT-PCR)was performed to amplify the gene of PS1TP5 using the mRNA of HepG2 cells as template and the gene was cloned into pGEM-T vector.The gene of PS1TP5 was cut from pGEM-T-PS1TP5 vector and cloned into yeast expressive plasmid pGBKT7,then pGBKT7-PS1TP5 was transformed into yeast cell AH109.The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western hybridization.Results PS1TP5 gene was successfully amplified and identified by DNA sequencing.The digested fragment was cloned into pGBKT7 vector and transformed into yeast cell AH109.The results of SDS-PAGE and Western assay showed that the relative molecular weight of the expressed product was about 36 950,and PS1TP5 protein existed in yeast cells.Conclusion The findings suggest that PS1TP5 can be successfully expressed in yeast cell.

OBJECTIVETo assess the value of dynamic contrast-enhanced MRI (DMRI) in predicting early response to neoadjuvant chemotherapy (NAC) in patients with locally advanced breast cancer (LABC) and to assess the accuracy of MRI in evaluation of residual disease after NAC.

METHODSForty-three women with LABC (44 lesions, all were invasive ductal carcinoma) underwent DMRI before, after the first and final cycles of NAC. For each patient, the tumor volume, early enhancement ratio (E1), maximum enhancement ratio (Emax), and maximum enhancement time (Tmax), dynamic signal intensity-time curve were obtained during treatment. The residual tumor volumes obtained by DMRI were compared with pathological findings to assess the accuracy of DMRI.

RESULTSAfter the first cycle of NAC, the mean volume of responders decreased insignificantly (P = 0.055), but after NAC, mean volume of residual tumor decreased significantly (P = 0.000). Morphological changes: 29 cases showed a concentric shrinkage pattern while 7 cases showed a dendritic shrinkage pattern. Significant differences were found in E1, Emax and Tmax between responders and non-responders (P < 0.05). After the first cycle of NAC, E1, Emax and Tmax of responders changed significantly (P < 0.001), while there was no significant change in non-responders (P > 0.05). After NAC, the dynamic signal intensity-time types were changed in responders, and tended to be significantly flattening, while no significant change was found in non-responders. The residual tumor volume correlation coefficient between MRI and pathology measurements was very high (r = 0.866, P < 0.01).

CONCLUSIONDMRI is useful to evaluate the early response to NAC in LABC. The presence and volume of residual tumor in LABC patients treated with NAC can be accurately evaluated by DMRI.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; drug therapy ; pathology ; Carboplatin ; administration & dosage ; Carcinoma, Ductal, Breast ; drug therapy ; pathology ; Chemotherapy, Adjuvant ; Contrast Media ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Staging ; Neoplasm, Residual ; Paclitaxel ; administration & dosage

OBJECTIVETo express and purify the HIV p24 gene in E. coli cells, and identify p24 antigen activity.

METHODSThe full length gene fragment of HIV p24 was amplified by PCR and inserted into the pRSET vector in order to construct the pRSET-p24 recombined vector. After transforming into E. coil, the purified p24 protein was prepared by metal-ligand affinity chromatography (IMAC). The accuracy of inserted gene and activity, specificity of HIV p24 proteins were detected by two enzymes digestion technology, SDS-PAGE, Western Blot (WB) and ELISA.

RESULTSThe length of the HIV p24 gene fragment was 690 bp after digesting the recombinant plasmid T-p24 and pRSET-p24 with BamH I and Hind III. The expressed proteins had a single expected band of about 24 x 10(3) in SDS-PAGE. The specificity and activity of p24 protein were tested by WB and ELISA.

CONCLUSIONThe HIV p24 sequence from HIV-1 gene plasmid hasbeen expressed in E. coil. This protein possessed good specificity and activity.

Blotting, Western ; Chromatography, Affinity ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Gene Expression ; HIV Core Protein p24 ; genetics ; isolation & purification ; metabolism ; Humans ; Plasmids ; genetics ; Polymerase Chain Reaction ; Recombinant Proteins ; isolation & purification ; metabolism