Objective To investigate the construction of compound membrane with corneal stromal cells and collagen-chitosan by tissue engineering technique and its biocompatibility.Methods Rabbit and human corneal stromal cells were separated and seeded into collagen-chitosan membrane.The compound membrane was transplanted into rabbit corneal stroma.Then the growth condition of keratocytes,the effect on normal keratocytes and degradation of compound membrane were detected by corneal confocal microscope,anterior OCT and histological and immunohistochemical methods ex vivo 1,2,4 weeks after grafting.Results The rabbit and human corneal stromal cells grown well in collagen-chitosan scaffold.The compound membrane degradated gradually after grafting.There was no necrosis and dissolvation.Corneal epithelium,stroma and endothelial cells were all normal.Conclusion Collagen-chitosan can be used as a biological scaffold for construction of corneal stroma.Corneal confocal microscopy and anterior OCT are new methods to observe the biological activity of constructed corneal stroma.

Objective To investigate the effects of GGNBP2 on proliferation,migration and invasion of human gli-oma U251 cells and the potential mechanism. Methods U251 glioma cells were transfected with lentiviral vector carrying GGNBP2 to establish a stable overexperssion of GGNBP2 in U251 cell line. After verification of the efficiency of transfec-tion by real-time PCR, Western Blot, PCR and CCK-8 were applied to detect the proliferation of U251 cells. The Tran-swell chamber assay was applied to measure the migration and invasion of human U251 glioma cells. Western blot was applied to measure protein levels of AKT, p-AKT, PCNA and MMP9. Result The stable U251 glioma cell line overex-pressing GGNBP2 was successfully established. Compared with the control group, overexpression of GGNBP2 significant-ly inhibited the proliferation(P<0.05),invasion(57±6 vs. 203±6,205±7,F=512.4,P < 0.05)and migration (74±7 vs. 254±14,248±13,F=242.5,P<0.05) of U251 cells. The protein expression levels of p-AKT (F=45.4, P<0.05), PCNA (F=348.5, P<0.05) and MMP-9 (F=88.7, P<0.05) in GGNBP2 group were markedly decreased compared with the control group. Conclusion GGNBP2 may suppress the proliferation, invasion and migration of glioma though down-regulation of p-AKT, which in turn inhibits PCNA and MMP-9 expression.

Vitreoscilla Hemoglobin(VHb), with the function of increasing the growth of and product yield by a heterologous host, has been widely use in the area of fermentation, environment protection, transgenic animal and plant, recombinant protein expression, etc. Fusion protein of VHb with other enzyme or protein can enhance activity and stability of the enzyme or isolation efficiency of the protein. The reconstitution of VHb will be helpful to obtain ‘novel’ proteins which have better activity.

ObjectiveTo investigate the correlation between glucose excursions and oxidative stress in the elderly type 2 diabetic patients. MethodsFifty five elderly type 2 diabetic patients (32 males and 23 females) were recruited. All the patients were submitted to continuous glucose monitoring system (CGMS) for three days. According to the result of CGMS monitoring ,the patients were divided into two groups: high glucose excursion group (30 cases) and low glucose excursion group (25 cases). In high glucose excursion group, the therapy was adjusted for 4 weeks based on the CGMS monitoring. After the treatments, the specimens of blood were collected again to detect the related indexes. Results(1) There were no differences in age, blood pressure, fasting blood glucose and glycosylated hemoglobin between the two groups (all P>0.05). The stardard deviation of blood glucose was higher, and the maximun and average amplitude of glycemic excursion were significantly lower in high glucose excursion group than in low glucose excursion group (t= 5. 4620,5. 9416,3. 8281, all P<0.05) ;(2) Compared with low glucose excursion group, plasma concentration of 8-iso-PGF2a was obviously higher in high glucose excursion group[(57.56 ± 3.86)ng/L vs.(34. 21±3. 82) ng/L, t= 18. 221, P=0. 0000)] . (3) Stepwise regression analysis showed that the standard deviation of blood glucose was involved in regression model (β= 1. 959, P= 0. 013).ConclusionsIn elderly patients with type 2 diabetes mellitus, glucose excursion is related with oxidation stress, which suggests that the glucose excursion may be the risk factor for oxidation stress.

Objective To investigate the expression of SEL1L and p63 protein in esophageal squamous cell carcinoma and precarcinomacous lesion.Methods Immunohistochemical staining(EnVision method)was employed to detect the expression of SEL1L and p63 protein in 60 samples of esophageal squamous cell carcinoma,32 samples of high grade esophageal intraepithelial neoplasia,13 samples of low grade esophageal intraepithelial neoplasia and 33 samples of normal esophageal mucosa.Results The positive rate of SEL1L protein expression was 61.5%in low grade intraepithelia neoplasia,90.6%in high grade intraepithelial neoplasia and 96.7%in esophageal squamous cell carcinoma,significantly higher than that in normal esophageal mucosa(6.1%)(P0.05).Conclusion Both the expression of SEL1L and p63 protein increases steadily in the progression of esophageal squamous cell carcinoma,which indicates that the two genes may play a role and cooperate with each other in the carcinogenesis.

[Summary] Thyroid-associated ophthalmopathy ocular damage involved in lacrimal, tear film, cornea, and conjunctiva which can lead to ocular discomfort and decreased visual.The mechanism of thyroid-associated ophthalmopathy ocular surface damage and treatment is reviewed.

Reversible cerebral vasoconstriction syndrome (RCVS) is characterized by thunderclap headache and reversible cerebral vasoconstriction, with other neurologic signs and symptoms. To the best of our knowledge, there were only a few cases of RCVS presenting both as both convexity subarachnoid hemorrhage (cSAH) and posterior reversible encephalopathy syndrome (PRES). Herein, we report a case of a 32-year-old woman with RCVS who presented with recurrent thunderclap headaches that occurred 50 days after delivery, with cSAH and PRES on magnetic resonance imaging (MRI). She had significant clinical and radiological recovery on 3 months’ follow-up. The clinical coexistence of cSAH and PRES in our case with RCVS is quite rare. This case illustrates the importance of awareness of the diagnosis of RCVS among clinicians even when initial brain and vascular imaging are normal. Early diagnosis and treatment are crucial for better prognosis.

Objective - ，，，，，- - - To establish a gas chromatographic method for the determination of trans 1 1 1 4 4 4 hexafluoro 2 ［ - （）］ Methods - （） butene HFO 1366mzz E in workplace air. HFO 1366mzz E in air was directly collected with aluminum foil ， ， ， composite plastic bag separated by dimethylpolysiloxane capillary column detected by flame ionization detector and Results - （） - 3， quantified with external standard method. The linear range of HFO 1366mzz E was 6.82 68 200.00 mg/m with the 3， correlation coefficient of 0.999 9. The detection limit and the lower limit of quantitation were 0.59 and 1.98 mg/m respectively. - - - The recovery rate was within 95.45% 103.05%. The relative standard deviation of within batch precision and between batch - - ， precision were 2.26% 5.07% and 4.09% 6.82% respectively. The samples can be stored at room temperature for at least seven Conclusion ， ， days. This method is simple to use with a wide linear range low detection limit high accuracy and precision and - （） good sample stability. It can be used for the detection of HFO 1366mzz E in the air of workplace

Objective To evaluate the curative effect of stress urinary incontinence (SUI) with Tconsling suspension surgery. Methods From May 2010 to September 2011,we treated 9 cases SUI with T-consling suspension surgery.The average age was 51 years (range 45 -63 years).They all suffered from stress incontinence 1 -5 years. Urodynamic test were performed preoperatively and stress urinary incontinence was confirmed.Abdominal leak point pressures is between 75 - 103 cm H2O (average 87 cm H2O),and no obvious cystocele vaginalis and others pelvic organ prolapsed disease were detected.Operation was done by transvaginal front wall incision to place T-consling suspension tape beneath the middle urethra.Results Operation time were between 15 to 25 mins,with 17 minutes on average.EBL was 5 - 30 ml.Urinary catheters were drawn 3 -4 d postoperatively.In 9 cases,8 were cured,one improved.The improved case had leakage when abdominal pressure increases,but became less significant,and the other cases had no leakage when abdominal pressure increases.The short-term cure rate was 89％. Conclusions The Tconsling suspension is safe,effective and easy to perform for the treatment of SUI,however,longer follow-up and more cases are necessary to confirm the long-term effects of this procedure.

Abstract: Objective - - To establish a pre column derivatization high performance liquid chromatography method for detecting Methods dimethyl sulfate (DMS) in workplace air. DMS in workplace air was collected with mercaptopyridine impregnated （ silicone tube. The derivative of DMS and mercaptopyridine was eluted by mobile phase phase A: water, phase B: acetonitrile, ∶ the volume ratio was 40 60) , and separated with a C18 column, then detected with diode array detector and quantitated by a Results - standard curve. The linear range of DMS was 0.17 40.00 mg/L, with the correlation coefficient of 0.999 95. The detection limit and the lower limit of quantitation were 0.05 and 0.17 mg/L respectively. The minimum detection concentration and minimum quantitation concentration were 0.02 and 0.04 mg/m³, respectively (air sample volume of 4.5 L, 1.0 mL sample - - - solution). The average desorption efficiency was 98.40% 102.00%. The within run and between run relative standard deviations - - were 0.61% 3.92% and 1.71% 6.00%, respectively. The samples could be stored at room temperature for at least 14 days. Conclusion This method can be used to detect DMS in workplace air.