Objective:To examine the expression and function of thrombonspondin-1 (TSP-1) in oxygen-induced retinopathy in new-born mice, and to investigate its role in retinal neovascularization. Methods:A total of 40 C57BL/6J newborn mice were divided equally into a model group (n=20)andanormalcontrolgroup(n=20).Miceinthemodelgroupwereexposedto75%oxygentoestablish the oxygen-induced retinopathy (OIR) model. On the 7th, 9th, and 11th day after the birth of mice, 5 mice were randomly selected each time from the 2 groups to examine the expression of TSP-1 mRNA with reverse transcription polymerase chain reaction (RT-PCR). After that, 5 mice were selected on the 11th day to observe the retinal neovascularization by fluorescein angiography retinal flatmount. Results:On the 11th day, fluorescein angiography retinal flatmount showed that the retinal blood vessels presented mean network distribution in the normal control group, while in the model group, a lot of dilatated areas in the retinal main vessels surrounded the optic disc. Meanwhile lots of new blood vessels were found surrounding the optic disc with irregular distribution but well distributed peripherial retinal small vessels, which was typical of early stage OIR. There was no signiifcant difference in the retinal TSP-1 mRNA level between the model group and the normal control group in the postnatal 7-day mice (P>0.05). Compared with the normal control group, the expression of TSP-1 mRNA in the model group was signiifcantly lower in postnatal 9-day and 11-day mice (P<0.05, P<0.01) , and the expression of TSP-1 mRNA in postnatal 9-day mice was lower than that in the postnatal 11-day mice(P<0.05).
Conclusion:In the early stage of OIR model (retinal vascular growth and development stage), the expression of TSP-1 mRNA in the retinal tissue is gradually decreased, implying that TSP-1 (as a negative regulatory factor) may be involved in the formation of retinal neovascularization in the early stage.

Objective: To investigate the effect and mechanism of rapamycin inhibiting mammalian target of RAPA (mTOR) on heart valve cell calciifcation in experimental rats. Methods: The rat’s valvular interstitial cells were isolated and the cells were cultured in 4 groups:①Normal control group,②Calciifcation group,③Rapamycin group and ④Calciifcation + rapamycin group. The apoptosis rates of valvular interstitial cells were detected by flow cytometry, calcium deposition was observed by Alizarin S staining, the calcified nodules were counted and the protein expressions of bmp-2, osteocalcin, osteopontin, smad-1 and caspase-3 were examined by Western blot analysis. Results: The rat's valvular interstitial cells were suceessfully isolated; the cell apoptosis rates were similar among different groups,P>0.05. The calciifed nodule in Calciifcation group (0.471 ± 0.091) was more than Normal control group (0.104 ± 0.023), while the nodule in Calciifcation + rapamycin group (0.237 ± 0.039) was less than Calciifcation group, allP<0.05. Compared with Normal control group, the protein expression levels of bmp-2, osteopontin and smad-1 were obviously increased in Calciifcation group,P<0.05, and compared with Calciifcation group, the above indexes were obviously lower in Calciifcation + rapamycin group, P<0.05. The protein expression levels of caspase-3 were similar among 3 experimental groups,P>0.05. Conclusion: Rapamycin may down-regulate the targeting protein expressions of bmp-2, osteopontin and smad-1 via inhibiting mTOR, therefore, reducing the valvular interstitial cell calcification which might be related to mTOR pathway suppression in experimental rats.

Objective:To explore the judging value of brain natriuretic peptide (BNP) on patient's condition of male patients with senile degenerated heart valvular disease (SDHVD) complicated heart failure (HF) .Methods :A total of 67 SDHVD + HF male patients were regarded as SHDVD group ,another 43 male HF patients ,who accepted treatment during the same period with corresponding age and without valvular heart disease ,were enrolled as pure HF group .Cardiac function indexes and plasma BNP level were measured and compared between two groups ,the correlation between BNP and left ventricular end -diastolic dimension (LVEDd) was analyzed ,and the cutoff point of BNP level for judging severity of SDHVD + HF was also calculated .Results:Compared with pure HF group , there were significant rise in LVEDd [ (49.79 ± 4.31) mm vs .(53.04 ± 7.10) mm] and plasma BNP level [ (214.43 ± 237.71) pg/ml vs . (682.06 ± 981.15 ) pg/ml ] , and significant reduction in left ventricular ejection fraction [LVEF ,(64.84 ± 5.83)% vs .(58.24 ± 7.99)% ] , P<0.05 or <0.01;percentage of moderate -severe heart dys‐function (NYHA class Ⅲ + Ⅳ) in SDHVD group was significantly higher than that of pure HF group (56.72% vs . 37.21% ) ,χ2 =3.988 , P=0.046. Spearman correlation analysis indicated that BNP level was positively correlated with LVEDd (r=0.588 ,P= 0.001) .The area under ROC (AUC) of BNP was 0.975 ,and the critical point of BNP level judging severity of male SDHVD complicated HF was 312 pg/ml ,and its sensitivity and specificity was 86.8% and 96.6% respectively .Conclusion:The SDHVD patient's condition is more severe compared with patients without valvular heart disease ,and BNP level is help to judging patient's condition .

Objective To study the application of spiral CT three-dimensional reconstruction in diagnosing intra-articular and other bone fractures in the earthquake of Wenchuan.Methods 25 cases with joint and other bone fractures in Wenchuan earthquake underwent multi-slice spiral CT volume scanning,the images were dealed with three-dimensional reconstruction at workstation including multiplanar reconstruction (MPR) and surface shaded display(SSD) to observe the fractures.Results 36 fractures in 25 cases were found by three-dimensional reconstruction(MPR and SSD),which included knee joint bone fracture in 6,one case was avulsion fracture of the point attached to the tibia by posterior cruciate ligament and missed by X-ray;ankle fracture in 3,1 case was Pilon fracture;shoulder fracture in 5,one case with glenoid anterior labrum missed by radiography;elbow fracture in 3;spinal fractures including cervical vertebra in 2,thoracic vertebra in 4 and lumbar vertebra in 5,2 cases were burst fracture accompanied with the articular process fracture; pelvis bone fracture in 8,1 case was sacroiliac fracture accompanied with sacroiliac separation.There were open bone fractures in 4 cases,fractures with dislocation in 6 cases,soft tissue swelling in different degree around the fractures in 25 cases.Conclusion Spiral CT three-dimensional reconstruction of bone can directly and really show the fractures that will provide in-formations for the clinical diagnosis and treatment .

AIM: To confirm the effects of capillary endothelium injury in the lung with ischemia reperfusion. METHODS: The rabbits pulmonary models of ischemia reperfusion (I-R)injury were established. Plasma nitric oxide and endothelin levels, arterial oxygen tension and wet/dry weight ratios were determined in different periods in control and I-R groups, and the pulmonary ultrastructure abnormities were analyzed under electromicroscope. RESULTS: The plasma levels of NO and ET-1 in I-R animals increased significantly ,compared with those in sham treated control groups. The level of ET-1 had significantly negative correlation with PaO 2 and positive correlation with the value of the wet/dry weight ratios. The swelling and karyopyknosis of capillary endothelium in the lung with I-R groups were observed in ischemia periods. The injuries of endotheliums and typeⅠandⅡ alveolar cells in 0 5 h of reperfusion were more severe than those in ischemia time, and the injuries began to repair in 2 h of reperfusion time. CONCLUSION: In the I-R group, pulmonary capillary endothelium have been injuried, which may play a prominent role in I-R injury and dysfunction of the lung.

Objective To investigate the characteristics of the dynamic contractions on the surface electromyography (sEMG) single of stroke patients induced from the low limb muscle when exercising passively, exercising initiatively with assistant and against resistance.Methods 24 stroke patients with hemiplegia and 17 normal subjects were tested with sEMG under a dynamic contractions in coxa and knee flexion and extension passively, initiatively with assistant and against resistance. The myoelectric signals were collected and processed by linear time and frequency domain method.Results The values of median frequency (MF) and mean power frequency (MPF) of stroke group were significantly lower, but the value of average EMG (AEMG) was higher ( P<0.001). The values of MF and MPF in activity side were lower than that in non-activity side ( P<0.001). The values of MF and MPF when exercising passively were higher than that when exercising with resistance ( P<0.05). The value of AEMG when exercising with resistance was highest. The values of MF and MPF in the synergist muscle were higher. The values of AEMG in the antagonistic muscle and synergist muscle were higher than that agonist and synergist muscle ( P<0.01). The values of MF and MPF in non-paretic exercising side were higher significantly, but in paretic exercising side and non-paretic silent side were lower. The values of MF and MPF in exercising side from vastus lateralis (VL) were the highest. The values of AEMG in exercising side and non-exercising side from biceps femoris (BF) were the highest. The values of MF and MPF in low limb of stroke group reduced, that in rectus femoris (RF) from paretic side was the lowest; that in BF from non-paretic side was the lowest ( P<0.01). The value of AEMG in low limb of stroke group was high significantly, especially in BF from the low limb of the non-paretic side in stroke patients. The values of AEMG in four group muscles gradually were higher following the higher exercising load, and that in the BF was the highest, and that in vastus medialis (VM) rose significantly.Conclusion The values of MF and MPF of stroke patients with hemiplegia reduce significantly, but the value of AEMG is higher. The values of MF and MPF in exercising side are lower than that in non-exercising side and non-paretic exercising side rising significantly, but that in paretic exercising side and non-paretic silent side reduce significantly. The values of MF and MPF in assistant exercise are higher than that in passive exercise and resistance exercise, but the value of AEMG in resistance exercise is higher than that in assistant exercise and in passive exercise. The values of MF and MPF in synergist muscle rise, but the values of AEMG in antagonist and synergist muscle are higher than that agonist and synergist muscle.

OBJECTIVETo study features of brain gray matter injury in cerebral infarction patients and intervention of scalp acupuncture by using voxel-based morphology.

METHODSA total of 16 cerebral infarction patients were recruited in this study, and assigned to the scalp acupuncture group and the control group, 8 in each group. Another 16 healthy volunteers were recruited as a normal group. All patients received scanning of T1 structure. Images were managed using VBM8 Software package. Difference of the gray matter structure was compared among the scalp acupuncture group, the control group, and the healthy volunteers.

RESULTSCompared with healthy volunteers, gray matter injury of cerebral infarction patients mainly occurred in 14 brain regions such as cingulate gyrus, precuneus, cuneus, anterior central gyrus, insular lobe, and so on. They were mainly distributed in affected side. Two weeks after treatment when compared with healthy volunteers, gray matter injury of cerebral infarction patients in the scalp acupuncture group still existed in 8 brain regions such as bilateral lingual gyrus, posterior cingulate gyrus, left cuneus, right precuneus, and so on. New gray matter injury occurred in lingual gyrus and posterior cingulate gyrus. Two weeks after treatment when compared with healthy volunteers, gray matter injury of cerebral infarction patients in the control group existed in 23 brain regions: bilateral anterior cingulum, caudate nucleus, cuneate lobe, insular lobe, inferior frontal gyrus, medial frontal gyrus, precuneus, paracentral lobule, superior temporal gyrus, middle temporal gyrus, lingual gyrus, right postcentral gyrus, posterior cingulate gyrus, precentral gyrus, middle frontal gyrus, and so on. New gray matter injury still existed in 9 cerebral regions such as lingual gyrus, posterior cingulate gyrus, postcentral gyrus, and so on.

CONCLUSIONSBrain gray matter structure is widely injured after cerebral infarction. Brain gray matter volume gradually decreased as time went by. Combined use of scalp acupuncture might inhibit the progression of gray matter injury more effectively.

Acupuncture Therapy ; Brain ; physiopathology ; Brain Injuries ; therapy ; Cerebral Infarction ; therapy ; Gray Matter ; pathology ; Humans ; Magnetic Resonance Imaging ; Scalp ; Stroke ; therapy

Background The multipotent differentiation features of induced pluripotent stem cells (iPSCs) offer a new option for cell replacement therapy of many clinical diseases.In ophthalmology,iPSCs are a good model in studying the pathogenic mechanism of degenerative ocular diseases.A better identification method for iPSCs is critical for analyzing the in vivo biological characteristics of iPSCs.Objective This study was to investigate the feasibility and stability of labeling iPSCs with quantum dots.Methods Human umbilical mesenchymal stromal cells-iPSC lines were cultured and amplified on matrigel,and the characteristics of iPSCs were evaluated by immunofluorescence.Different concentrations (5.0,7.5 and 10.0 nmol/L) of quantum dots with a CdSe/ZnS nuclear shell structure were used to label iPSCs after passaging and proliferation.The labeling outcome was observed with a three-dimensional deconvolution real-time live cells imaging system.The labeled iPSCs were subsequently cultivated,and then changes in fluorescence intensity were examined 7 days after the first and the second passaging of iPSCs.Results iPSCs were observed to grow in a clonal manner under the inverted microscope.The iPSC markers,OCT4 and Nanog,were detected by immunofluorescence.With increasing concentrations of quantum dots,the fluorescence intensities representing the levels of OCT4 and Nanog in iPSCs were gradually elevated,with optimal levels of fluorescence observed at a concentration of 10 nmol/L of quantum dots.The fluorescent labeling of OCT4 and Nanog in iPSCs remained and weakened gradually till day 7 even after the second passage.Conclusions Quantum dots labeling could be used to track iPSCs in a dose-independent manner.The fluorescent signal from the quantum dots labeling the iPSCs lasts 2 weeks at least.

OBJECTIVETo introduce an arthroscopic technique in managing recurrent dislocation of the patella and its clinical results.

METHODSSixteen patients with recurrent patellar dislocation were reviewed, including 3 males and 13 females. The average age was 17.6 years old (ranged from 14 to 32 years). The patients suffering from patellar sub-luxation averaged 18.5 months (ranged from 6 to 23 months)before operation. These patients were treated with lateral patellar retinacular release outside the synovial bursa of knee joint and medial patellofemoral ligament reconstruction using the semitendinosus tendon free autograft. The Lyshohm scores before and after operation were used to evaluate outcomes at the final follow-up.

RESULTSAll the patients were followed up, and the duration ranged from 6 to 48 months, with an average of 12 months. There was no recurrence. The Q angle decreased from (16.4 ± 3.7)° to (10.1 ± 1.4)°; insall index decreased from 1.37 ± 0.25 to 1.28 ± 0.23; congruence angle decreased from (21.3 ± 2.6)° to (5.86 ± 2.23)°; Lysholm score improved from 76.1 ± 5.2 to 89.8 ± 4.1 at 6 months after operation.

CONCLUSIONCompared with conventional procedure, arthroscopic surgery for recurrent dislocation of the patella achieves excellent outcomes with minimum invasion.

Adolescent ; Adult ; Arthroscopy ; Bursa, Synovial ; surgery ; Female ; Humans ; Knee Joint ; surgery ; Male ; Patellar Dislocation ; physiopathology ; surgery ; Patellar Ligament ; surgery ; Range of Motion, Articular ; Treatment Outcome ; Young Adult

Background The construction of tissue-engineered corneal endothelium needs the functional seeding cells,so how to culture a large amount of functional corneal endothelial cells (CECs) is an urgent problem to be solved.Objective The aim of this study was to evaluate the role of aqueous humor on bovine CECs in vitro.Methods Aqueous humor of 1.2 ml was collected from the anterior chamber of bovine and sterilized,and the liquid supernatant was obtained.The bovine CECs were isolated from bovine cornea and then cultured in low glucose Dulbecco Modified Eagle Medium with 10％ fetal bovine serum (FBS) in vitro.Aqueous humor was added into the medium with the final concentration of 2.5％,5.0％,l0.0％,15.0％ and 20.0％,respectively,and no aqueous humor was added in the control group.Cell counting kit-8 (CCK-8) assay was used to detect the absorbency value of CECs for the evaluation of cell proliferation.Progression of the cell cycle was analyzed by flow cytometry (FCM).After confluence of the cells was reached,1 ml plastic spear tip was used to scratch the cell single layer,and the cells were incubated consequently in medium with 10％ FBS and with or without aqueous humor for 24 hours.Healing area of the cell single layer was measured.The cells were incubated at a density of 6 × 105 cells/ml and cultured using medium with or without 10.0％ aqueous human for 5 days,and the number of the cells was analyzed by DAPI fluorescence technique.Results Under the phase-contrast microscopy,the confluent CECs showed a slabstone-like and hexagonal appearance.CCK-8 assay revealed that the absorbance values of CECs was significantly different among the various culture groups (F=4.051,P =0.007),and the absorbance value in different concentrations of aqueous human culture groups was significantly higher than that in the control group (P ＜ 0.01).FCM showed that the percentage of the cells in S-G2 phases was (34.80-±3.13)％ in the 10.0％ aqueous humors group and (23.06±1.13)％ in the control group,showing a significant difference (t =-5.729,P=0.005).The scratch test showed that the healing area of the cell signal layer was (0.116±0.019) mm2 in the 10.0％ aqueous humors group and (0.358 ±0.049) mm2 in the control group,showing a significant difference (t =13.842,P =0.000).The density of cells in the 10.0％ aqueous humor group was (1439± 1 10)/field,which was more than (1162±45)/field in the control group (t =-11.020,P=0.000).Conclusions Aqueous humor at the concentration of 10.0％ promote the growth and proliferation of bovine CECs.The result suggests that 10.0％ aqueous humor can be used as a promoting agent during the culture of CECs.