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.

BACKGROUND:After knee joint replacement, patients are often treated with Rivaroxaban and Enoxaparin Sodium for postoperative anticoagulation, avoiding the formation of deep vein thrombosis in lower limbs. OBJECTIVE:To explore the application effects of different anticoagulant drugs in patients with knee joint replacement. METHODS:Ninety patients underwent knee joint replacement in Xiangya Second Hospital of Central South University from July 2011 to July 2014, were randomly divided into two groups, with 45 patients in each group. The experimental group was treated with Rivaroxaban, while the control group was treated with Enoxaparin. RESULTS AND CONCLUSION:Postoperative drainage volume, total blood transfusion, bleeding index, quantity of blood platelet, activated partial thromboplastin time, thrombin reduction time, whole blood viscosity, plasma viscosity, D-dimer coagulation index, HSS score at postoperative 2 weeks, average ecchymosis area, average thigh circumference, and average leg circumference were significantly better in the experimental group, than in the control group (P<0.05). The incidence of deep vein thrombosis of lower limbs in the experimental group was lower than that in the control group (P<0.05). Experimental findings indicate that, both Rivaroxaban and Enoxaparin can exert anticoagulation effect during the knee joint replacement, and Rivaroxaban is better.

Objective To investigate the role of cyclophilin A (CypA) and CD147 in the process of skin aging. Methods Twenty cases of tissue samples from junior group(＜15 years old), middle age group(30-40 years old)or old age group (＞65 years old) were collected from photophobic and exposal parts of skin, respectively. Immunohistochemistry (IHC) and in situ hybridization (ISH) were carried out to semi-quantitatively detect the expression level of CyPA and CD147. Results IHC demonstrated that both CyPA and CD147 were expressed in both photophobic and exposal parts of normal human skin in all 3 groups. The expression levels of both CyPA and CD147 were increased with increase in age. There were significant differences in both CyPA and CD147 expression among 3 groups (P<0.05). CyPA and CD147 were also positively correlated in all 3 groups. Similar results were achieved by ISH. Conclusion The interaction between CD147 and CyPA might play an important role in the process of skin aging.

Improving medical students' calculation ability in statistics has become the focus and difficulty of medical statistics course teaching, and its application relies heavily on statistical calculation software. In order to explore a new teaching approach which combined the advantages of traditional method and web-based calculation, we intended to build a web computing platform applying the Browser/Server (B/S) mode based on the campus network, and to revise the current syllabus of medical statistics, as well as to create a virtual web-lab containing a typical case library. Practice has proved that the new mode effec-tively improved the practice capability of students and changed the traditional teacher-centered teaching approach.

Objective To investigate the expression and the possible clinical significance of serum Golgi protein(GP73)in infantile hepatitis syndrome(IHS)by different causes.Methods Totally 79 patients with IHS in Guangzhou Women and Children's Medical Center from February 2012 to December 2012 were enrolled in this study,including 15 cases with biliary atresia(BA)group,29 cases with infection(infection group),5 cases with neonatal intrahepatic cholestasis caused by citrin deficiency(NICCD group),and 30 cases with unknown etiology(idiopathic infantile hepatitis group).At the same time,30 healthy infants were enrolled as healthy control group.The serum levels of GP73 were determined by quantitative enzyme-linked immunosorbent assay(ELISA),and the children's liver function[alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),direct bilirubin(DBIL),alkaline phosphatase(ALP),γ-glutamyl trans-peptidase(γ-GT),total bile acid(TBA)and albumin(ALB)] were measured by turbidimetric inhibition immuno assay.Then,the corresponding data were statistically analyzed.Results Serum GP73 in BA group,infection group,NICCD group,idiopathic infantile hepatitis group and the healthy control group were(296.6±67.5)μg/L,(185.1±66.4)μg/L,(199.2±87.1)μg/L,(181.7±74.2)μg/L and(65.3±17.0)μg/L,respectively.Serum γ-GT levels in BA group,infection group,NICCD group,idiopathic infantile hepatitis group and healthy control group were(764.7±775.8)U/L,(448.2±352.7)U/L,(239.4±88.7)U/L,(283.3±377.2)U/L and(54.0±72.6)U/L,respectively.The levels of GP73 and γ-GT were significantly higher in infants with IHS,and the levels of GP73 and γ-GT in infants with BA were the highest(F=46.775,9.238,all P<0.05).There was a positive correlation between serum levels of GP73 and γ-GT(r=0.280,P<0.05),and there was no correlation between GP73 and ALT,AST,TBIL,DBIL,ALP,TBA,ALB(r=-0.061,-0.071,0.164,0.123,0.137,0.008,-0.047,respectively,all P>0.05).The receiver operating characteristic curve(ROC)constructed with GP73 showed a sensitivity of 80.0％and specificity of 82.8％with an area under the receiver(AUC)of 0.872 for diagnosis of BA,comparatively,a sensitivity of 66.7％and specificity of 71.9％were showed with a AUC of 0.731 when performed with γ-GT.Conclusions Serum GP73 concentration significantly increased in all liver disease groups,regardless of the etiology.Serum GP73 expression is significantly higher in infants with BA.Serum GP73 shows a superior sensitivity and specificity to γ-GT for diagnosis of BA,which might be useful for early diagnosis of BA and IHS with different causes.

The key to maximize the sensitivity of inner filter effect ( IFE)-based assay is to enlarge the overlap between the absorption spectra of the absorber and the excitation or emission spectra of the fluorophore. In this work, Mn-doped ZnS quantum dots ( QDs) were chosen for IFE-based detection of β-glucuronidase ( GUS) , since the excitation of QDs was perfectly overlapped with the absorption of the substrate of GUS, namely 4-nitrophenyl-β-D-glucuronide ( PNPG) . In addition, the phosphorescence emission from Mn-doped ZnS QDs could eliminate the fluorescence background from biological samples. Therefore, a simple turn-on phosphorescent GUS assay was developed, with the linear range of 10-300 U/L and limit of detection of 7 U/L (S/N=3).

italic>Artemisia argyi (A. argyi) is a Chinese herbal medicine in China. The main active components are volatile oils, flavonoids, and other compounds, which have various pharmacological activities. Methoxylated flavonoids are the main active ingredients in A. argyi. Flavonoid O-methyltransferase (FOMT) is a key enzyme in the O-methylation of flavonoids. In order to further understand the function and characteristics of FOMT proteins, this paper carried out the whole genome mining and identification of FOMT genes in A. argyi and performed phylogenetic, chromosomal localization, gene sequence characterization, subcellular localization prediction, protein structure, gene structure analysis, and expression pattern analysis. The results showed that a total of 83 FOMT genes were identified in the genome of A. argyi. The phylogenetic tree shows that FOMT genes are divided into two subgroups, CCoAOMT (caffeoyl CoA O-methyltransferase) subfamily (32 genes) and COMT (caffeic acid O-methyltransferase) subfamily (51 genes). Gene sequence analysis showed that the number of amino acids encoded by FOMT was 70-734 aa, the molecular weight was 25 296.55-34 241.3 Da, and the isoelectric point was 4.51-9.99. Compared with 32 members of the CCoAOMT subfamily, nearly 1/3 of the 51 members of the COMT subfamily were hydrophobic proteins and 2/3 were hydrophilic proteins. Subcellular localization prediction showed that more than 80% of CCoAOMT subfamily members were located in the cytoplasm, and 96% of COMT subfamily members were located in the chloroplast. COMT subfamily members have more motifs than CCoAOMT subfamily members. The N-terminal motifs of COMT subfamily proteins are relatively variable, while the C-terminal motifs are relatively conserved. Expression pattern analysis showed that CCoAOMT subfamily members were mainly expressed in roots, while COMT members were mainly expressed in leaves. Some FOMTs showed the tissue expression specificity by real-time quantitative PCR analysis, especially in leaves. In this study, we identified and analyzed the FOMT gene family in A. argyi, and provided a theoretical basis for further research on the function of FOMTs and the biosynthesis of methylated flavonoids in A. argyi.

Objective To assess the series of special anti-fatigue foods including JuntiⅠ, JuntiⅡ, JuntiⅢ, and Jun-tiⅣon military physical performance capacity .Methods Fifty-four soldiers , selected from a border defense troop , were randomly designated to control group , trial group 1 and trial group 2.Subjects of two trial groups were supplied with No .1 nutritional package ( including JuntiⅠ, Ⅲand Ⅳ) and No.2 nutritional package ( including Junti Ⅱ, Ⅲand Ⅳ),re-spectively, while no additional nutritional supplements were added in control group .After 7 days’ supplementation, a hard military exercise was performed to induce fatigue and an increasing load test was used to assess physical activity .RPE scale, exhaustive time and time taken to reach the 75% maximal heart rate were recorded while serum markers , such as glucose, lactate, BUN, LDH,and CK, were detected after test .Moreover, serum lactate and fatigue recovery scale were determined on the evening of the same day and the next morning .Results Prolonged exhaustive time and time taken to reach the 75%maximal heart rate and elevated RPE scores at 6 min were detected in both two trial groups compared with the control group .Meanwhile , after the increasing load test , elevated glucose concentration and reduced lactate , BUN, LDH and CK were also observed in both trial groups .Moreover, serum lactate of both trial groups was quickly recovered on the evening of the same day compared with the control group , and the next morning , serum lactate was even much lower in trial groups than in control group .The fatigue recovery scores were higher in trial groups at both time points .Meanwhile, there was no difference of such indexes between the two trial groups .Conclusion Through the combination use , the series of special anti-fatigue foods, inclucling No.1 and No.2 nutritional packages , can significantly improve the soldiers′physi-cal performance capacity , delay the physical fatigue emergence , promote physical activity recovery and prevent military training injury.

OBJECTIVE: To investigate the role of Human MutL homologue 1 (hMLH1) gene promoter methylation in the occurrence and development of osteoarthritis (OA). METHODS: General DNA was dealt with sodium bisulfite. The methylation of hMLH1 promoter was detected by methylation-specific PCR (MSP). hMLH1 protein expression in joint cartilage was detected by immunohistochemical method. RESULTS: The positive percent of hMLH1 promoter methylation in OA patients was higher than that in healthy persons (χ(2)=30.634, P<0.001); the positive percent of hMLH1 protein in OA patients was significantly lower than that in healthy persons (χ(2)=37.724, P<0.001); prmoter methylation and protein expression level of hMLH1 gene showed negative correlation (rs=-0.554, P<0.001). CONCLUSION hMLH1 promoter is hypermethlated in joint cartilage cells of OA patients. Hypermethylation may affect the protein expression of hMLH1, which might play a role in the occurrence and development of OA.
Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Adolescent ; Adult ; Cartilage, Articular ; metabolism ; Child ; DNA Methylation ; Female ; Humans ; Male ; MutL Protein Homolog 1 ; Nuclear Proteins ; genetics ; metabolism ; Osteoarthritis ; genetics ; metabolism ; Promoter Regions, Genetic ; genetics ; Young Adult

Objective To report the phototoxicity effects of a novel photosensitizer ZnPcS4-BSA on photodynamic therapy (PDT) towards human U251 glioma cells in vitro. Methods The cellular uptake of ZnPcS4-BSA by U251 glioma cells was quantified by UV-spectra to determine the optimal incubation time. Human U251 glioma cells were incubated with ZnPcS4-BSA of various concentrations and received laser irradiation of different energy densities. Cell survival rates were measured by CCK-8 assay.Flow cytometer was used to detect apoptosis.Gene expressions of vascular endothelial growth factor (VEGF) were detected by Real-Time PCR in the U251 cells after PDT and β-actin was used as an internal standard. The normal U251 cells severed as controls. Results The uptake of ZnPcS4-BSA by U251 glioma cells reached the maximum after incubation for 4 hours.ZnPcS4-BSA of different concentrations without laser irradiation had no significant effects on cell survival rates (P＞0.05).Without ZnPcS4-BSA incubation,compared with 0,25,50,100,200 J/cm2 groups, the cell survival rate of the 400 J/cm2 group was significantly lower (P＜0.05), whereas no significant difference was found between any other two groups. When the U251 glioma cells incubated with 30 μ mol/L ZnPcS4-BSA for 4 hours underwent laser irradiations of 25,50,100,200 J/cm2,the cellular survival rates significantly decreased with the increased energy densities (P＜0.05). When the U251 glioma cells incubated with ZnPcS4-BSA of 20,40,60,80,100 μ mol/L for 4 hours underwent laser irradiation of 200 J/cm2, the cellular inhibition rates significantly increased with the increased concentrations (P ＜0.05). Compared with controls, the cellular apoptosis and VEGF expression significantly increased in the U251 glioma cells incubated with ZnPcS4-BSA of 20 μmol/L after laser irradiation of 100 J/cm2 (P＜0.05). Conclusion The novel ZnPcS4-BSA is a good photosensitizer for PDT towards U251 glioma cells,because the ZnPcS4-BSA-mediated PDT can induce effective apoptosis of the targeted cells.