Objective To observe the changes of intracerebral amino acid transmitters in the periventricular leukomalacia (PVL) of newborn rats with microdialysis and so as to explore the role of excitotoxicity in PVL.Methods Replicated the model for PVL at the age of postnatal day 5 (P5) by intracerebral injection of 3-nitropropionic acid (3-NPA).Before injection of 3-NPA,and 15 min,30 min,45 min,60 min,75 min,90 min after injection of 3-NPA,collected the sample of extracellular fluid (ECF) at the corpus callosum above the left ventricle through microdialysis,respectively.After microdialysis,the experimental rats were allowed to survive to P6-P14,and then they were killed and the brains were prepared for HE stain.The amino accids of dialysate were quantified through high performance liquid chromatography(HPLC),and then the excitotoxic index (EI) was calculated.Results Fifteen min to 45 min after injection of 3-NPA,the concentrations of glumate (Glu) and aspartic acid (Asp) of ECF elevated significantly,and then returned to the normal levels.Fifteen min to 75 min and 15 min to 30 min after injection of 3-NPA,the concentrations of glycine (Gly) and GABA significantly elevated,respectively,and returned to normal levels at 90 min and 45 min after injection of 3-NPA,respectively.But the EI,which indicated the balance of excitatory amino acids(EAAs) and inhibitory amno aciols(IAAs),significantly elevated 15 min to 75 min after injection of 3-NPA,then retured to normal level after 90 min.Sub-cortical and periventricular white matter rarefaction and significant lateral ventricle enlargement were observed in HE staining.Conclusions Changes of intracerebral amino acid transmitters in the PVL of newborn rats show regularity:EAAs,IAAs of ECF and EI elavate in the early stage,and then return to the normal level quickly.It indicates that excitotoxicity play a great role in PVL,especially at the early stage.Therefore,the preventions of PVL must be executed at the early stage.

Background The incidence of myopia is gradually increasing,and how to choose a better corrective method of myopia for the best visual demand is very important.Objective This study was to compare visual quality of implantable collamer lens (ICL) implantation with femtosecond laser in situ keratomileusis (FSLASIK) for moderate and high myopia using double-pass optical quality analysis system (OQAS).Methods A non-randomized controlled clinical trail was performed.Fifty-two eyes with-4.00 to-9.00 D of 26 consecutive patients were included in NO.1 Hospital of Xi'an from January 2015 to January 2016.Twenty-four eyes of 12 patients with the corneal thickness ＜500 μm received ICL implantation as ICL group and 28 eyes of 14 patients which corneal thickness was ≥500 μm underwent FS-LASIK surgery as FS-LASIK group.The demography was matched between the two groups (all at P＞0.05).All the patients were followed-up for 3 months after surgery.The preoperative best corrected distance visual acuity (BCDVA),spherical equivalent (SE),postoperative uncorrected distance visual acuity (UCDVA),BCDVA and SE were examined and compared between two groups.The parameters from OQAS were evaluated and intergrouply compared,including the objective scattering inders (OSI),modulation transfer function (MTF) cut off frequency,Strehl ratio and OQAS values under the contrast of 100％,20％ and 9％ (OV100,OV20,OV9).Results The postoperative BCDVA was not significantly different from preoperative UCDVA in both ICL group and FS-LASIK group (-0.04±t0.10 vs.0.05±0.12;-0.07±0.12 vs.0.00±0.12) (t=3.128,2.358,both at P＞0.05).No statistically significant differnces were found in SE,UCDVA and BCDVA after operation between ICL group and FS-LASIK group (t =1.292,0.900,-0.653,all at P＞0.05).OQAS examination showed that MTF cut off,OSI,Strehl ratio,OV100 were not significantly different after operation between ICL group and FS-LASIK group (t=-2.032,-1.440,-0.224,all at P＞0.05).The postoperative OV20 and OV9 were 0.82±0.14 and 0.80±0.21 in the ICL group,which were significantly higher than those in the FS-LASIK group(0.59±0.15 and 0.47±0.13) (t =4.105,4.702,both at P＜0.05).Conclusions Both ICL implantation and FS-LASIK provide good optical and visual quality for moderate to high myopic eyes,and ICL appears to have a better visual quality in comparison with FS-LASIK under the contrasts of 20％ and 9％.

Objective To study the differentiation and proliferation ability of the spinal neural stem cells (NSCs) at different gestational ages in fetal rats. Methods Sprague-Dawley fetal rats were divided into group A (12 days of pregnancy), group B (14 days of pregnancy) and group C (16 days of pregnancy). NSCs were separated with enzyme-assisted microdissection. The diameter and numbers of NSCs balls were measured at different time. The cell growth curve was drawn with CCK8 colorimeter. NSCs were identified with BrdU/Nestin immunohistochemical staining. They were induced with 10% fetal bovine serum for 10 days, and the expression of β-tubulinⅢ and glial fibrillary acidic protein was detected with immunocytochemistry. Results There were cells expressed BrdU, Nestin, β-tubulinⅢ and GFAP in all the group. The most cells (22.74±0.79%) expressed β-tubulinⅢ in the group B, but no significant difference between group B and group C. The cell vitality on the 5th day of third-generation neural stem cells was the most in group B. Conclusion For enzyme-assisted microdissection, it may obtain more neurons to isolate the neural stem cells from 14 days of pregnancy pregnant rats.

Objective To investigate the inhibitory effect of aspirin on liver metastasis of colon cancer in mice and the possible mechanism.Methods A total of 32 BALB/C mice were injected with CT26 colorectal cancer cells to establish colon cancer liver metastatic model,with 3 mice dead,15 mice in control group and 14 mice in experimental group.The control group was given saline 0.2 mL/d,the experimental group were given aspirin 30 mg/kg.The liver weight and the number of metastatic tumors were calculated after 30 days of intervention.HE and CD31 staining was performed by immunohistochemistry to observe the metastasis and angiogenesis.The protein expression of VEGF and cox-2 were analyzed by Western blot.Results The average liver weight and number of liver metastases nodules in the experimental group were significantly lower than those in the control group(P<0.05).Pathological examination showed that the experimental group of mice the number of liver cells and liver tumor angiogenesis were significantly less than the control group(P<0.05).Western blot showed that the expression of VEGF and cox-2 of CT26 cells were down-regulated after treated with aspirin.Conclusion Aspirin can down regulate the protein expression of VEGF and cox-2 protein to inhibit liver metastasis of colon tumor proliferation and angiogenesis,thereby inhibiting metastasis of colon cancer cells,for therapeutic purposes.

Objective To construct the typeⅢ secretion system (T3SS) deficient mutant of O157∶H7 EDL933, and to detect its ability of attachment after infection with HeLa cells.Methods The recombinant DNA fragments obtained kana-mycin resistant gene were constructed by overlap extension PCR and transferred into EDL933/pKD46 to replace escR gene by homologous recombination.Results The T3SS-deficient mutant (ΔescR) was successfully construted.Compared with the wild-type EDL933, the growth rate and attachment on HeLa cells of ΔescR were significantly decreased.Conclusion The deletion of escR gene, which encodes T3SS structural protein EscR, affects the attachment of EDL933, suggesting a better basis for future studies of T3SS.

BACKGROUND:Surgical treatment for pigmented vil onodular synovitis can maximize the excision of synovial lesions and recovery of joint function.
OBJECTIVE:To investigate the knee joint function and relapse rate fol owing treatment of pigmented vil onodular synovitis with total knee replacement and arthroscopic synovectomy.
METHODS:We retrospectively analyzed 34 cases of knee pigmented vil onodular synovitis admitted for surgical treatment in the Department of Orthopedics, Xiangya Hospital of Central South University, China from December 2006 to December 2011. In these cases, 24 patients received arthroscopic synovectomy and 10 patients were subjected to total knee replacement. Adjuvant radiotherapy was conducted according to postoperative patient’s conditions. Lysholm scoring was employed in the arthroscopic synovectomy group, and American Knee Society scoring was used in the total knee replacement group. Knee function in two groups was compared before and after treatment. Fol ow-up observation was performed to compare knee function recovery and relapse rate between two groups.
RESULTS AND CONCLUSION:Thirty-four patients were effectively fol owed up for 12-66 months, mean 41.3 months. Statistical analysis showed that in the arthroscopic synovectomy group, the postoperative Lysholm score was (86.3± 10.3) points, significantly higher than the preoperative score which was (55.5±13.2) points (t=3.81, P=0.016, P<0.05). In the total knee arthroplasty group, the American Knee Society score was increased from (40.7±2.2) points preoperatively to (90.2±1.1) points postoperatively (t=6.27, P<0.01). In the arthroscopic synovectomy group, the American Knee Society score was increased from (34.2±3.9) points preoperatively to (80.8±1.9) points postoperatively (t=16.58, P<0.01). Arthroscopic synovectomy combined with adjuvant radiotherapy can achieve better outcomes in pigmented vil onodular synovitis patients, while the total knee replacement for advanced with advanced knee diffuse pigmented vil onodular synovitis is better to restore knee function and the recurrence rate is low.

Objective To evaluate the performance of the ABX Micro C-reactive protein(CRP)in determination of CRP.Methods The analytic characteristics including precision,carry-over,linearity, stability,interference and comparability were examined.Results The coefficient of variation(CV)was less than 5.1%,10% and d.3% for within-run,between-run and between-day,respectively.Carryover was less than 1.2%.Whole blood samples held at either room temperature or 4℃ were stable for 48 hours with relative deviation less than 6.0% relatively.Linear range was 1.0-70.0 mg/L using undiluted samples.The comparison between the ABX Micro CRP and Behring Nephelometer Ⅱ was well correlated Both serum:Y=0.996 7X-0.398 5,r~2=0.965 9;serum for BN Ⅱ,whole-blood samples for the ABX Micro CRP:Y=0.908 8X-0.138 2,r~2=0.959 4;both serum and whole-blood samples for the ABX Micro CRP: Y=1.001 7X-0.898 2,r~2=0.952 7.No obvious interference was observed by hyperhemoglobinemia and hyperlipidemia.Conclusion The determination of CRP test with ABX Micro is accurate and reliable.

The glucoamylase gene (glaA) of Aspergillus niger CICIM F0410 was cloned, sequenced and expressed. The integrated plasmid pBC-Hygro-glaA carrying the glaA was constructed and transformed into A. niger F0410. Transformants with multiple copies of glaA integrated in the chromosome were selected by 150 microg/mL hygromycin B and identified by real-time PCR. Two to three multiples of glaA in the chromosome were found to be optimal for higher expression of glucoamylase. Shake-flask fermentation under optimal conditions showed that glucoamylase secreted by the transformant GB0506 was 17.5% higher than parental strain F0410 at the end of fermentation. In conclusion, increasing copy number of glaA by chromosomal integration significantly improves the yield of glucoamylase in the industrial strain of A. niger.
Aspergillus niger ; genetics ; Cloning, Molecular ; Glucan 1,4-alpha-Glucosidase ; biosynthesis ; genetics ; Plasmids ; Recombinant Proteins ; biosynthesis

Objective To construct a prokaryotic plasmid expressing the recombinant protein of enterohemorrhagic Escherichia coli(EHEC) effector NleB1 and to prepare the polyclonal antibody of mouse anti-NleB1.Methods The nleB1 (990 bp) gene was amplified from the genome EHEC O157∶H7 and cloned into the expression plasmid pET24a to construct the recombinant plasmid pET24a-nleB1 that was transformed into E.coli BL21(DE3).After induction with isopropylthio-gelactoside( IPTG) , the His-tag fusion proteins were purified by Ni+affinity chromatography and gel slices.The polyclonal antibody was prepared by immunizing BALB/c mice with purified recombinant proteins and analyzed by Western blotting and ELISA.Results The pET24a-nleB1 recombinant plasmid was successfully constructed, the fusion protein was ex-pressed and purified,and the polyclonal antibody was obtained by immunizing mice with purified fusion protein.Western blotting and ELISA staining demonstrated that the polyclonal antibody was successfully obtained.Conclusion The prepara-tion of the polyclonal antibody against EHEC O157∶H7 NleB1 will be of help for further studies on the function of NleB1 protein.

Serum cortisol and plasma ACTH were determined in 10 patients with severe sepsis and 12 with septic shock on day 1,3,5 after diagnosis were made,and the data were compared with 12 control patients. To evaluate the hypothalamic-pituitary-adrenal(HPA)axis function in patients with severe sepsis and septic shock,1?g ACTH stimulation test was applied after hormone concentrations were obtained on day 1.Compared with the control patients,ACTH level was significantly higher in patients with severe sepsis and lower in septic shock(P