AIM: To explore the feasibility of culturing human umbilical vein endothelial cells ( HUVEC ) on acellular corneal stroma and performing the posterior lamellar endothelial keratoplasty ( PLEK ) treating corneal endothelial decompensation.
METHODS: Thirty New-Zealand rabbits were divided into three groups randomly, 10 rabbits for experimental group, 10 for stroma group and 10 for control group. Corneal endothelial cells were removed to establish animal model of corneal endothelial failure. PLEK was performed on the rabbits of experimental group and stroma group, and nothing was transplantated onto the rabbits of control group with the deep layer excised only. Postoperative observation was taken for 3mo. The degree of corneal edema and central corneal thickness were recorded for statistical analysis.
RESULTS:Corneas in experimental group were relieved in edema obviously compared with that in stroma group and the control group, and showed increased transparency 7d after the operation. The average density of endothelial cells was 2 026. 4±129. 3cells/mm2 , and average central corneal thickness was 505. 2±25. 4μm in experimental group, while 1 535. 6±114. 5μm in stroma group and 1 493. 5±70. 2μm in control group 3mo after operation.
CONCLUSION:We achieved preliminary success in our study that culturing HUVEC on acellular corneal stroma and performing PLEK for corneal endothelial decompensation. HUVEC transplanted could survive in vivo, and have normal biological function of keeping cornea transparent. This study provides a new idea and a new way clinically for the treatment of corneal endothelial diseases.

Objective To explore the effect of the mutant and expression level of N-ras on chronic myelogenous leukemia. Method We investigated the mutant by direct sequencing in a K562 human chronic myelogenous leukemia cell line, with determination of the expression level of N-ras mRNA in K562 by RT-PCR. Result No single point mutation was detected in K562 cell line, furthermore, the expression level of N-ras gene is abnormaly high in contrast to normal human. Conclusion Our results indicated that the expression level of N-ras gene was obviously high in K562 cell line, and the underlying mechanism was not only mutation, so that further investigation is called for.

Objective To explore the relationship of gallbladder emptying function and its hemodynamics in diabetic patients. Methods Gallbladder volume ,emptying function and its arterial blood flow were measured by ultrasound. Results The gallbladder volumes of fasting (FV ) and postprandial (RV) increased and the ejection fraction (EF) of gallbladder was decreased in DM group than in NC group (P<0.01). In DM group ,the gallbladder artery’s resistance index (RI) was higher than in NC group [(0.61 ± 0.06) vs (0.70 ± 0.08)](P<0.01). Minimum diastolic blood flow velocity (Vmin) was lower than in NC group [(8.72 ± 1.43) vs (6.05 ± 1.89)cm/s](P<0.01). RI was associated with HbA1c (r= 0.827 ,P< 0.01) and LDL‐C(r= 0.820 ,P< 0.01). Conclusion Gallbladder emptying dysfunction in diabetic patients is associated with the hemodynamics of gallbladder.

Magnetic resonance (MR) images can be used to detect lesions in the brains of patients with multiple sclerosis (MS). An automatic method is presented for segmentation of MS lesions using multispectral MR images in this paper. Firstly, a Pd-w image is subtracted from its corresponding T1-w images to get an image in which the cerebral spinal fluid (CSF) is enhanced. Secondly, based on kernel fuzzy c-means clustering (KFCM) algorithm, the enhanced image and the corresponding T2-w image are segmented respectively to extract the CSF region and the CSF-MS lesions combinatoin region. A raw MS lesions image is obtained by subtracting the CSF region from CSF-MS region. Thirdly, based on applying median filter and thresholding to the raw image, the MS lesions were detected finally. Results were tested on BrainWeb images and evaluated with Dice similarity coefficient (DSC), sensitivity (Sens), specificity (Spec) and accuracy (Acc). The testing results were satisfactory.
Algorithms ; Brain ; pathology ; Humans ; Image Enhancement ; Magnetic Resonance Imaging ; Multiple Sclerosis ; diagnosis ; Sensitivity and Specificity

Background Bevacizumab has been widely used in the treatment of new blood vessel disease in ophthalmology.The investigation of the pharmacokinetics and safety after intracameral injection of bevacizumab can offer the basis for the management of iris neovascularization and neovascular glaucoma.Objective The present study was to observe the distribution of bevacizumab(avastin)in eye tissue and toxic effects following the injection of anterior chamber.Methods Twenty-four New Zealand albino rabbits were divided into two groups randomly.0.05 ml (1.25mg)of Bevacizumab was intracamerally injected into the left eyes in the experimental group,and a balanced salt solution of 0.05 ml was injected in the same way into the left eyes of the control group.The anterior segment of eyes and ocular fundus were examined by slit-lamp microscope and direct ophthalmoscope after injection.Intraocular pressure was measured and corneal endothelial microscopy was performed before and after the injections.Five rabbits of the two groups were sacrificed on the first day,the fourth day,the seventh day,the fourteenth day,and the thirtieth day after injection,and the eyeballs were enucleated for histopathological examination.The ultrastructure of eye tissue was observed under the transmission electron microscope on the fourth day and the thirtieth day,and then immunofluorescence staining were performed to assess the distribution of bevacizumab in the eye tissues.This experiment complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission(Version 1988).Results No abnormality in the cornea,lens,vitreous and retina was observed after the injection of bevacizumab under the slit lamp microscope and direct ophthalmoscope.No significant differences were found in intraocular pressure and corneal endothelial cell density in the bevacizumab group compared with the control group before injection and 2 hours,1 day,7 days,14 days,30 days after injection(P =0.760,P =0.956).No histopathological and ultrastructural changes of the cornea,lens,chamber angle,iris,ciliary body and retina were seen after the injection in the experimental group and control group under the light microscope and transmission electron microscope.Bevacizumab was distributed in the anterior chamber angle,iris,ciliary body,choroid and retina in injected eyes and fellow eyes after intracameral injection with red fluorescence and presented the dynamic changes with the lapse of time.The immunofluorescence response of eye tissue to bevacizumab was weaker in the fellow eyes compared with injected eyes.Bevacizumab was mainly distributed in the vessel wall and lumen.Conclusions Bevacizumab can quickly distribute in the vascular tissue of the anterior chamber angle,iris,ciliary body,choroid and retina in injected eyes after intracameral injection without obvious toxic effects to eye tissue.Bevacizumab administered intracamerally may be a new strategy or a joint strategy for iris neovascularisation.

Objective To investigate the effects of Puerarin on glucose and lipid metabolism and gastric motility in early period type 2 diabetic (T2DM) rats.Methods Rat model of T2DM was established by high fat-sugar diet fed and low-dose streptozotocin-treated. SD rats were divided randomly into normal control group (NC), normal+Puerarin group (NP), diabetes control group (DC) and diabetes+Puerarin group (DP). NP and DP rats were given Puerarin 400 mg/(kg?d) once per day for 5 weeks, NC and DC rats were given PBS. Half time of gastric emptying and emptying rate were evaluated by SPECT. The serum level of FBG, GSP, FFA, TC, TG and INS were measured by kit.Results Compared with NC group, DC rats had higher FBG, FFA, TC, GSP, TG and emptying rate, but INS and half time of gastric emptying decreased significantly (P<0.05,P<0.01). Compared with DC group, TG, GSP, FFA and emptying rate of DP rats were reduced (P<0.05), but had more half time of gastric emptying (P<0.05). The results of multivariate stepwise regression analysis showed that FBG related to half time of gastric emptying.Conclusion Type 2 diabetic rats have faster gastric motility, higher blood glucose and lipid. Puerarin might improve the disorders of GSP, TG, FFA and gastric emptying in diabetic rats.

Objective:To compare two methods for the determination of compound cod-liver oil emulsion. Methods:The content of ciprofloxacin in compound cod-liver oil emulsion was determined by HPLC and UV, respectively. The determination by HPLC was performed on a Thermo C18 column (150 mm ×4.6 mm, 5 μm). The mobile phase was 0.025 mol·L-1phosphate-acetonitrile (87∶13)andpHwasadjustedto3.0±0.1withtriethylamine. Thedetectionwavelengthwas277nmandtheflowratewas1.5ml·min-1. The detection wavelength of UV was 277 nm. Results:The average recovery of HPLC and UV was 100. 44% and 100. 84% with RSD of 1. 01% and 1. 09% (n=9), respectively. The detection results of the two methods were compared by paired sample t-test, and no statistically significant difference was found (P>0. 05). Conclusion:The two methods are specific and accurate, and can be used for the determination of ciprofloxacin hydrochloride in compound cod-liver oil emulsion.

Objective To investigate the effect of diaphragm training on respiratory function and the activities of daily living after cerebral apoplexy.Methods Seventy-eight patients with post-stroke fatigue were randomly divided into a treatment group and a routine therapy group,each of 39.Both groups were given conventional rehabilitation treatment,while the treatment group was additionally provided with diaphragm muscle training.Before and after 4 weeks of treatment,both groups were evaluated using pulmonary function tests,a fatigue severity scale (FSS),Fugl-Meyer motor function assessment (FMA) and the modified Barthel index (MBI).Results Vital capacity,forced vital capacity,forced vital capacity during the first second,and especially the maximum ventilatory volume of the treatment group were all significantly improved after the treatment.There was also significant improvement in that group's average FSS score,but not in that of the control group.The average FMA and MBI scores for both groups had improved significantly after the intervention,with the average FMA score of the treatment group significantly higher than that of the control group.However,no significant differences in the average MBI score were observed after the treatment.Conclusion Diaphragm training can significantly improve motor function and the daily life of stroke survivors.The mechanism may be related to improved respiratory function and decreased severity of fatigue.

Objective: To discuss the clinical efficacy of bandage contact lens (BCL)in pterygium excision combined with conjunctival transplantation for the older adults,and to explain its advantages.Methods:The clinical materials of 70 older patients (70 eyes ) who underwent the pterygium excision combined with conjunctival transplantation were retrospectively analyzed.They were divided into two groups according to the postoperative treatment.In the observation group,35 cases (35 eyes)wore BCL after operation.In control group,35 cases (35 eyes)were covered with normal eye pad.The scores of blurred vision,photophobia,foreign body sensation, lacrimation,eye-soreness,corneal fluorescence stain (FSC)and epithelial defect areas were observed at 1,3,7 and 10 d afer operation.Results:The scores of blurred vision,photophobia,foreign body sensation,lacrimation,eye-soreness of the patients in observation group were significantly lower than those in control group at 1,3,and 7 d afer operation (P <0.05).The scores of FSC and epithelial defect areas of the patients in observation group were significantly lower than those in control group at 3 and 7 d afer operation (P <0.05).There were no significant differences in blurred vision,photophobia,foreign body sensation,lacrimation,eye-soreness,FSC and epithelial defect areas of the patients at 10 d afer operation between two groups (P >0.05).No complications of infection and implant healing badness were observed in all patients.Conclusion:BCL can significantly reduce the irritations after pterygium excision combined with conjunctival transplantation for the older adults and promote the healing of the corneal epithelium.

Background Diabetic retinopathy (DR) is a common ocular complication of diabetes,and its pathogenesis is associated with a variety of factors.c-Jun N terminal kinase (JNK),one of the genes involving in apoptosis,plays an important role in the pathology of diabetes,and relative research is catching increasing interests in recent years.Objective This study was to quantify the expression of JNK3 in retinas of DR murine.Methods Forty-eight SPF male C57BL/6 mice were randomly divided into the diabetes group and the normal control group.Diabetic mouse models were establishend by intraperitoneal injection of 1％ streptozocin (STZ) dissolved by sodium citrate buffer,and equvilant volume of sodium citrate buffer was used in the same way in the mice of the control mice.The left eyeballs were obtained 2,4,8 weeks after modeling and the retinas were collected.Real-time quantitaive PCR was perfored to detect the expression of JNK3 mRNA in retinas.The use and care of the experimental mice complied with the Administration of Experimental Animals in Kunming Medical College.Results Blood glucose levels were significantly higher in 2,4,8 weeks after modeling in the diabetic group compared with the normal control group (t=-5.675,-5.498,-5.347,all at P＜0.01).The relative expression levels of JNK3 mRNA (A value) in the retinas were significantly different between the groups at various time points (Fgroup =102.345,P＜0.05 ; Ftime =131.679,P＜ 0.05).The relative expression levels of JNK3 mRNA in the retinas were 3.21 ±0.14 and 5.43 ±O.37 in 4 and 8 weeks after modeling in the diabetic group,which were significantly elevated in comparison with the normal control group (2.54±0.42 versus 2.26±0.67) (t =4.073,23.399,both at P＜0.05).Compared with the second week and fourth week,the relative expression levels of JNK3 mRNA in the retinas in the eighth week were significantly raised in the diabetic group (t =10.756,16.857,both at P ＜ 0.05).Conclusions JNK3 expression in the retina upregulates in diabtic mice in a time-dependent manner.JNK3 is paopably involved in the pathogenesis and development of DR.