Objective To investigate the effect of indomethacin (IND)-loaded micelles on rheumatoid arthritis and its mechanism. Methods Freund’s-induced adjuvant arthritis and carrageenan-induced acute arthritis in SD rats were employed to evaluate the therapeutic effect of IND-loaded polymeric micelles after intraarticuar drug administration. Fifty-six SD rats were randomly divided to receive IND-loaded micelles at a dose of 4.5, 1.5, 0.5 mg/kg respectively, IND injection, oral administration of IND, blank PNIPAm/EAB-PPP-1 micelles at the 8~ th day once a day for a week after subcutaneous injection of 0.1 ml Freund’s into voix pedis of the right hind foot. The normal control rats took normal saline instead of Freund’s. There were 8 rats in each group. Results IND-loaded micelles could inhibit the swollen joint. No rats of intraarticular drug administration developed ulceration. Conclusion The sustained therapeutic efficacy could be achieved through intraarticular injection of IND-loaded micelles. Local delivery of IND can avoid severe gastrointestinal stimulation that frequently results from oral administration of IND.

BACKGROUND: It is important to study the methods of culturing bone marrow-derived mesenchymal stem cells (MSCs) to obtain a great amount of high purity MSCs for applying ocular tissues constructed by tissue engineering technique to treat eye diseases.OBJECTTVE: To separate and culture in vitro MSCs from bone marrow of the adult rats by density gradient centrifugation combined with adherence culture, and observe the growing characteristics and the possibility of mass multiplication.DESIGN: A completely randomized grouping design/repetitive measuring experiment.SETTING: Pathological laboratory, Zhongshan Ophthalmic Center, Sun Yat-sen University.MATERIALS: Four six-week-old SD rats about 250 g, grade Ⅱ of cleaning, were provided by the Animal Center of Sun Yat-sen University [certificate number: SCSK(Yue)2004/0011], about 250 g each rat and there was no limit to the sex. The main reagents and instruments included low sugar Dulbecco modified Eagle culture medium (DMEM/F12, American Gibco Corporation), trypsin (fetal bovine serum (FBS, Hangzhou Sijiqing Bio-Engineering Material Research Institute), American Gibco Corporation), disodium edetate, lymphocyte separating medium, fibronectin, CD44, CD34, CD31 monoclonal antibodies, two-step-method kit for immunohistochemistry (Beijing Zhong shan Biotechnology Corporation).METHODS: This experiment was conducted at the Key laboratory of Ophthalmology (Sun Yat-sen University), Ministry of ethanol (750 g/L) for 10 minutes. Under aseptic condition, the medullaris cavitas was exposed, the syringe containing application m edium was directly punctured into the femoral cavity, the cells in the medullaris cavitas were washed out with the culture medium containing heparin and taken as the cell suspension. The bone marrow-derived MSCs were separated and purified by density gradient centrifugation combined with adherence culture, and the growing conditions of the wells. When the cells had generally connected with each other, they were fixed with methanol or dimethy ketone in situ for 10 minutes, and then hematoxylin eosin (HE) staining or immunohistochemical staining. Antibodies against fiinoculated to 96-well culture plate by a cell density of 4.25×107/L, with 200 μL every well; then put the culture plate into culture box. Then from the next day to the sixth day, 5 g/L MTT solution was added into two rows (20 μL every well) every day, continuously cultured for 4 hours, then the supernatant was removed, and 200 μL DMSO was added to each well, agitated for 5 minutes, then detected the absorbance (A) values at 570 nm wave length, and a growth curve was drawn.branes were clear and the cell bodies were lucent. Being cultured for 2 days, there appeared adherent cells. 3 days later,most of the adherent cells extended and appeared to be in polygon, star or long-shuttle shapes. 4 days later, the cells showed to be in division growth stage; and about 12 days later, the cell clones were connected to each other, appearing curve of subcultured MSCs was in S shape. Cells began growing fast on the 2nd day after being passaged, and they entered the growth period for 3-4 days followed by saturation period, and then cells stopped growing.CONCLUSION: It is a simple and practical method to separate MSCs from the bone marrow of adult rats by means of density gradient centrifugation and adherence. MSCs can greatly proliferate in vitro and offer seed cells for the application of tissue engineering technique to treat eye diseases.

Objective To evaluate the value of the fusion of 99m Tc-MIBI imaging technology apply in the diagnosis of malignant lung tumor. Methods Thirty - four cases with lung nodules proved by X-ray and/or CT scanning, a total of 48 lung nodular lesions. And the imaging with-99m Tc-MIBI of chest performed at 10 minutes and 2 hours delayed after injection by GE Infinia Hawkeye 4 SPECT-CT. then the regions of interesting ( ROI) were drawn in the tumor and contra lateral position to calculate the radioactivity ratios of tumor to normal ( T/NT) , and fused with the spiral CT scanning image in the same machine, and reading the early and delayed image respectively. Judged the result of the image develops, and statistical analysis of the ratio (T/NT) according to the final pathologic consequence. Results All cases with total of 48 nodular lesions, 21 nodules were positive in early imaging, 16 nodules were positive in delayed imaging (the ratio T/NT over 3. 33). defined the delayed image positive as the final criterion, The(T/NT)ratios of Malignant lung lesions were significantly higher than the benign lesions ( P ＜0. 05). Negative nodes 27, 13 cases of lung cancer lesions were malignant, confirmed by postoperative pathologic examination. The falsepositive nodules 3, false-negative nodules 2. The sensitivity was: 88.88%, the specificity was: 90.9% positive predictive value ( +PV) was: 84. 21% , negative predictive value (-PV) is: 93.75%. Conclusion 99mTc-MIBI as a tumor positive imaging agent is highly sensitivity to lung lesions, but specificity is not so high.

ObjectiveTo investigate the effect of intensive rehabilitation training on gross motor function in children with cerebral palsy. Methods101 children with cerebral palsy were divided into routine group (n=51) and trunk group (n=50). They were assessed with Gross Motor Function Measure (GMFM) before and after treatment. ResultsThe scores of GMFM improved in both group after treatment. There was significant difference between them 6 months after treatment (P<0.05), but not within 3 months (P＞0.05). ConclusionIntensive rehabilitation training can improve the recovery of gross motor function in children with cerebral palsy.

Background The pathological basis of human primary open angle glaucoma mainly is the degeneration of trabecular meshwork and Schlemm canal.As the outflow pathway of aqueous humor, the tissue origin and characteristics of trabecular meshwork remain less clear.Studying the characteristics of trabecular meshwork may contribute a new thought to the treatment of primary open angle glaucoma.Objective This study was to explore the histological property of trabecular meshwork by detecting the expression of D2-40, a lymphatic biomarker,and CD31,CD34 and smooth muscle actin (SMA), some vascular endothelial biomarkers.Methods Twenty specimens of adult eyeballs were collected from Zhongshan Ophthalmic Center of Sun Yat-sen University,including l0 eyeballs from accident death donors,3 enucleated eyeballs due to posterior segment of choroidal malignant melanoma and 7 orbital exenterated eyeballs.The series ocular meridian sections with the thickness of 4 μm were prepared for the haematoxylin-eosin staining.Immunohistochemistry was employed to detect the expression of D2-40,CD31 ,CD34 and SMA in trabecular meshwork and Schlemm canal.Results Intact structure of chamber angle was observed under the optical microscope by haematoxylin-eosin staining.Human trabecular meshwork tissue was presented with meshlike structure,while Schlemm canal showed the lumen structure.No abnormal substance and periphery synechia were found.D2-40 was strongly expressed in the trabecular meshwork endothelial cells with staining score for 3 points, but CD31, CD34 and SMA were negatively expressed.In contrast, CD31, CD34 and SMA rather than D2-40 were positively expressed in the Schlemm canal.In addition, CD31 and CD34 were also positively expressed in the vascular endothelial cells around the trabecular meshwork.Conclusions The trabecular meshwork expresses lymphatic biomarker rather than vascular biomarkers.This result indicates that trabecular meshwork of human eyes is lymphatic vessel-like tissue.

Objective To analyze the economic effects of different rehabilitation patterns for children suffering from cerebral palsy. Methods A total of 153 cerebral palsy patients were divided into a hospital-community-family rehabilitation group(n = 52), a hospital rehabilitation group (n = 50) and a non-intervention control group (n = 51). Those in the first group were provided with a hospital-community-family rehabilitation therapy pattern, those in the sec-ond only hospital rehabilitation and the third no intervention. All the patients were evaluated using the Gross Motor Function Measure-88 (GMFM-88) Scale and the Cost Measure Scale at admission, and at the end of the 3rd and 6th months of treatment. Results There were no significant differences in gross motor function among the three groups at admission. At the end of the 3rd month and the 6th month there were significant differences between the children in the hospital-community-family rehabilitation program and those in the hospital rehabilitation program in terms of gross motor function. Their general percentage, monthly percentage and monthly relative percentage results were all significantly different. But there was no significant difference in the non-interventian control group since admission. Every unit of improvement in gross motor function cost $101.87±97.59, $75.11±45.75 in the hospital-community-family reha-bilitation program and $387.21±54.76, $170.31±123.16 in the hospital rehabilitation program at the end of the 3rd and the 6th month respectively. So the cost of the former was only about 30% of the latter. Conclusion Hospital rehabilitation is suitable for the early rehabilitation of cerebral palsy children. Hospital-community-family rehabilitation is better for long-term rehabilitation of cerebral palsy children, and what is more, it can decrease the rehabilitation ther-apy cost substantially. So a hospital-community-family rehabilitation pattern is more compatible with China's national situation.

Aim TostudytheroleofASIC1aonthe matrix turnover and MAPK expression of the rat articu-lar chondrocytes with extracellular acidosis. Methods ArticularchondrocyteswereisolatedfromSprague-Dawley rats, and their phenotype was determined by toluidine blue and immunocytochemical staining. The GAG content of cell culture supernatant was deter-mined by dimethyl-methylene blue spectrophotometric assay, while Hyp content by chloramine T assay. ELISA assay was used to measure MMP-2 , TIMP-2 content. Furthermore, the ERK1/2, p38 MAPK phos-phorylation protein expression levels were tested by Westernblotassay.Results ASIC1acontributedto the effect of GAG, Hyp and TIMP-2 levels reduction induced by extracellular acidification, while the effect of MMP-2 was weaker. Moreover, ASIC1a could in-crease the ERK1/2 , p38 MAPK phosphorylation pro-teinexpressionlevels.Conclusion ASIC1acould regulate rat articular chondrocytes matrix turnover via ERK1/2 and p38 MAPK signaling pathway, and there-by inhibit the rat articular cartilage damage induced by acidosis.

OBJECTIVETo investigate endostatin gene therapy of rat corneal neovascularization induced by acid cauterization.

METHODSpBlast-hEndostatin and pBlast-Mcs were identified by digestion with Nhe Iand Sal I, by PCR reaction, by sequence, and then by alignment of PCR products with the gene Bank using NCBIBLAST software. They were then purified with QIAGEN Endofree plasmid maxi kit. Rat corneal neovascularization models were made with 75% AgNO(3) and 25% KNO(3) cauterization. The treatment method was subconjunctive injection of the pBlast-hEndostatin with the control of pBlast-Mcs.

RESULTSpBlast-hEndostatin was found to contain the human endostatin gene. The rat corneal neovascularization induced by acid cauterization was significantly suppressed after subconjunctive injection of the pBlast-hEndostatin with inhibition rates of 37%, 40.2%, and 42.8% respectively on the sixth, tenth, and fifteenth day. The inhibition rate for the density of corneal neovascularization was 40%. However, no inhibition effect on the length of the neovascularization and corneal inflammatory cells was observed. Corneal neovascularization areas were positively correlated with edema and corneal opacity.

CONCLUSIONSThe plasmid of pBlast-hEndostatin contained the human endostatin gene. The rat corneal neovascularization induced by acid cauterization can be partially inhibited by subconjunctive injection of the pBlast-hEndostatin mediated by liposomes. Endostatin produced by transfected fibroblast cells directly inhibits corneal neovascularization. This is not caused by inflammatory reaction inhibition.

Animals ; Corneal Neovascularization ; pathology ; therapy ; Endostatins ; genetics ; Genetic Therapy ; methods ; Humans ; Rats ; Rats, Wistar ; Transfection

Objective:To analyze retrospectively the risk factors for pulmonary infection after traumatic cervical spinal cord injury.Methods:The 154 patients with a cervical spinal cord injury studied included 120 with a pulmonary infection and 34 uninfected controls. Regressions were evaluated using data on their genders, ages, the cause of injury, affected segments, the neurological level of the injury (NLI), and the presence of a vertebral fracture or dislocation.Results:Age, complete injury, NLI at C 1 to C 4, and an injury-to-treatment time of more than 8 hours were found to be independent risk factors for secondary pulmonary infection. Conclusion:Elderly spinal cord injury patients, with a complete injury, an NLI between C 1 and C 4 or an injury-to-treatment time of more than 8 hours have a higher risk of pulmonary infection.

Objective To investigate the safety and clinical efficacy of magnetic resonance neuronavigation assisted technique in neurosurgical clipping of pericallosal aneurysms. Methods From January 2010 to January 2017,40 consecutive patients with pericallosal aneurysm treated with neurosurgical clipping at the Department of Neurosurgery,the 175th Hospital of PLA were enrolled retrospectively. They were diagnosed by CT angiography (CTA),magnetic resonance angiography (MRA)or digital subtraction angiography (DSA)before operation. According to the different surgical methods,40 patients were divided into either a routine surgical group (n＝18)or a neuronavigation assistance group (n＝22). On the basis of the conventional longitudinal fissure approach,the neuronavigation assistance group was treated with the magnetic resonance neuronavigation technique. The aneurysms and upper drainage vein,design incision and surgical approaches were accurately located. The operation time,surgical complications (edema or infarction after drainage vein injury and secondary bleeding in the operated area)and proportion of good prognosis (the modified Rankin scale [mRS]score ＜3)were compared. Results (1)Under the microscope,40 patients underwent clipping of pericallosal aneurysms via longitudinal fissure approach. Postoperative CTA or DSA confirmed that they were all completed clipped. The operation time of the neuronavigation assistance group were shorter than that of the routine surgical group (2. 5 ± 0. 5 h vs. 3. 5 ± 0. 4 h,t＝1. 254),and the proportion of edema or venous infarction was less than that of the routine surgical group (4. 5%[1／22]vs. 6／18). The difference between the two groups was statistically significant (all P＜0. 05);there were no significant differences in the incidences of accidental rupture and secondary hemorrhage between the two groups (all P＞0. 05). (2)Both groups of patients completed the 6-month follow-up. There were 12 patients (12／18)with good prognosis in the routine surgery group and 20 (90. 9%)with good prognosis in the neuronavigation assistance group. There was no significant difference in the proportion of good prognosis between the two groups (χ2＝3. 545,P＞0. 05). Conclusions The use of magnetic resonance neuronavigation assisted technique helps the precise intraoperative positioning of the lesions and surgical approach optimization,thereby effectively implementing brain protection,reducing the risk of microsurgery, and improving the accuracy and safety of the surgery. It is an effective auxiliary means of neurosurgical clipping of pericallosal aneurysms.