Background Choroidal neovascularization(CNV)has been descrihed as a main reason of visual loss in a lot of ocular diseases.Researches showed that local hypoxia and retinal pigment epithelial(RPE) cells play an important role in the formation of CNV.A closely relationship of angiopoietin-2 (Ang-2) and angiogenesis has been proved.However,whether the expression of Aog-2 in hypoxic cultured human RPE cells is associated with the pathogenesis of CNV is still below understood.Objective This study was to investigate the effects of hypoxia on expression of Ang-2 in cultured human RPE cells in vitro,and discuss the possible effects of Ang-2 in the formation of CNV.Methods Human RPE cells were cultured and passaged,and 4-7 generation of cells were used in the experiment.The cells were incubated in cultural plate at the density of 5×107 cells/L.The culture medium containing 200 μmol/L CoCl2 was used to establish the hypoxia model of human RPE cells culturecd in vitro for 0.5,1,2,4,6,12and 24 hours,and the RPE cells cultured under normoxia were as controls.Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of Ang-2 mRNA in cultured human RPE cells,and enzymelinked immunosorbnent assay(ELISA) was used to assay the content of Ang-2 protein in supernatant of cultured human RPE cells.Results The survival rate of human RPE cells was 90％ after resoscitation.The fourth generation of cells showed the fusiform with the less pigment in them.The Ang-2 mRNA/β-actin mRNA value in human RPE cells was significantly different among various groups(F=1086.30,P=0.00),The Ang-2 mRN A/β-actin mRNA value in hypoxia cultured for 0.5 hours group began to increase and peaked in hypoxia culture for 4-6 hours group,with the significant differences in comparison with normoxia control group(P＜0.05).The Ang-2 mRNA/β-actin mRNA value decreased to the baseline level at hypoxia for 24 hours.The ELISA analysis showed that the concentration of Ang-2 protein in supernatant of RPE cells showed significant difference among groups(F=1034.00,P=0.00).The concentration of Ang-2 protein increased at hypoxia culture for 0.5 hours and peaked at 6 hours,showing significant differences in comparison with the control group (P＜0.05).Conclusions Hypoxia could significantly up-regulate the expression of Ang-2 in human RPE cells cultured in vitro.Ang-2 expresses highly in the early stage of hypoxia,implying that Ang-2 participates in the formation of CNV.

NDUFS3 is an essential subunit of mitochondrial NADH:ubiquinone oxidoreductase (complex Ⅰ ) and plays a critical role in the mitochondrial typeⅠ respiration chain.Mutations in this gene are shown to cause neurodegenerative disease such as Leigh syndrome (subacute necrotizing encephalopathy).In recent years,many evidences show that the expression of NDUFS3 proteins are lower in many cancerous cells compared to the corresponding normal cells.It comes to the conclusion that NDUFS3 may play a role in the tumorigenesis.

Objective To evaluate the biocompatibility between bone morphogenetic protein(BMP) composition and bone marrow stem cell (BMSc).Methods Microspheres were prepared with chitosan and sodium alginate, and BMP was enwrapped in the microspheres. The biocompatibility of the composition was examined using cell-culturing method. The BMSc was cultured in combination with microspheres. The extending speed of the cells, the proliferation and alkaline phosphatase activity were tested.Results There were no inhibition on cellular proliferation of BMSc when it was cultured in combination with microspheres in vitro, but ALP activity increased significantly.Conclusion BMP microspheres possessed satisfactory biocompatibility, and could increase the osteogenic capability of BMSc in vitro.

Objective To evaluate the value of ultrasonography in the diagnosis of pediatric ovarian torsion (OT).Methods Ultrasonography was performed in 38 OT,and 52 non-OT children.Bilateral ovarian area was measured.The larger area was defined as S1,whereas,the smaller area was defined as S2.S1/S2 ratio was calculated.Follicles around ovaries,ovarian blood flow,and pelvic effusion were also observed during the ultrasonographic scanning.ROC curve was employed to evaluate the clinical value of ultrasonography in pediatric OT diagnosis.Results ①S1/S2 ratio,incidence of follicles around ovaries and of no ovarian blood flow were all higher in OT compared to non-OT children (P =0.000).While pelvic effusion showed no significant difference between the two groups (P =0.004).②Using S1/S2 ratio to diagnose OT,the area under the ROC curve was 0.925 (0.865-0.986),the best cutoff point was 1.6,sensitivity was 97％ and specificity was 88％.Patients were divided into two groups based on the cutoff point of 1.6,the area under the ROC curve of S1/S2 was 0.924(0.847-0.969),of follicles around ovarian was 0.896(0.781-0.931),and of no ovarian blood flow was 0.911 (0.831-0.961),indicating S1/S2 held the highest diagnostic value.Conclusions Ultrasonography is an accurate method in the early diagnosis of pediatric ovarian torsion,with S1/S2 ratio holding the highest diagnostic value in this application.

Purpose:To evaluate the effect of Auto-PBSCT combined with long-time low-dose interleukin-2 (after transplant) for increasing the rate of 5-year disease-free survival of lymphoid malignancies (ALL and NHL).Methods:U- sing chemotherapy combined with low dose G-CSF to mobilize peripheral blood stem cells,we carried out auto-PBSCT in lymphoid malignancies (7 cases CR_1 ALL、1 case CR_2 ALL、3 cases CR_1 NHL、5 cases refractory NHL).After transplant, 15 cases used low dose interleukin-2 at 5.10~5U/d for one year or longer.Results:7 cases CR_1 ALL are still alive,the aver- age disease-free survival is 73 months.Median disease-free survival is 51.5 months.2 of 3 cases CR_1 NHL are still alive. The 6 years disease-free survival in 10 cases CR_1 lymphoid malignancies(CR_1 NHL and CR_1 ALL) is 0.90?0.11,the aver- age disease-free survival is 66.8 months.In another PR (refractory) group of NHL and ALL (CR_2),3 years disease-free survival was 0.33?0.18,and average survival time was 20.66 months,but a chemotherapy-sensitive NHL is still alive (63 months),a case of lymphoblast cell NHL also survived for 46 months and died after relapse.Conclusions:Auto-PBSCT combined with long-turm low dose IL-2(after transplant) could effectively increase the rate of 5-year disease-free survival in CR_1 lymphoid malignancies(ALL and NHL).

Objective To estimate an automatic DNA extracting method, and evaluate the quantity and quality of these DNA samples.Methods Genomic DNA was extracted automatically from blood samples by MiniPrep 75-Ⅱ workstation. The yield and purity of DNA samples were detected by UV-spectrophotometer, and the integrality of these DNA samples were measured by agarose gel electrophoresis.Results The yield of genomic DNA extracted from 100?l whole blood were (7.33?2.58)?g. The average purity of these DNA samples was 1.647?0.135 (A260/A280) and the molecular weight of them was about 21 kb.Conclusion High quality and intact genomic DNA can be extracted rapidly from whole blood by using MiniPrep 75-Ⅱ workstation, and these DNA samples were suitable for downstream molecular biology experiments.

AIM: To observe the clinical effects of intravitreal injection of triamcinolone acetonide ( TA ) before vitrectomy for retinal detachment associated with choroidal detachment. · METHODS: Totally 23 cases ( 23 eyes ) of retinal detachment associated with choroidal detachment in our hospital were treated by intravitreal injection of TA 4-5d before 23-Gauge micro-invasive vitrectomy combined with silicone oil injection. All the cases were followed up between 6 to 9mo. The anatomic retinal reattachment, visual acuity, intraocular pressure and postoperative complications were observed and analyzed. ·RESULTS: After the surgery, the visual acuity of all patients were improved, with 9 eyes better than 0. 3 (39%), and 18 eyes better than 0. 05 (78%). The BCVA at 1wk, 1 and 3mo and last follow up were different compared with before operations (P<0. 05). The mean intraocular pressure was 4. 02±1. 47mmHg before injection, 13.69±4. 68mmHg before operation (P<0. 05), and17.72±5.87 mmHg after operation (P<0.05). The retina of all patients treated were reattached 2wk post-operatively. The retinal reattachment rate after the primary surgery and the secondary surgery was 87% and 100%, respectively. Post-operative complications included 7 eyes of transient high intraocular pressure, occurred during 12-14d after operations and returned to normal after less glucocorticoid eye drops and giving IOP lowering drugs. There were no intraocular hemorrhage, iatrogenic retinal breaks, infections, or lens injuries. · CONCLUSION: Intravitreal injection of TA before vitrectomy for retinal detachment associated with choroidal could improve the clinical effects, and decrease the difficulty of surgery while the injection itself is pretty safe.

Objective To detect serum protein spectrum in gastric cancer, screen the serum proteins related to gastric carcinoma and build auxiliary diagnosis model with SELDI-TOF-MS, Methods The serum proteomic fingerprints were detected by SELDI-TOF-MS and CM10 proteinchip. The sera came from 50 first visit gastric cancer patients who were diagnosed by pathological method and 16 healthy peo- ple. The data were analyzed by Biomarker Wizard Software and the proteomic fingerprint of gastric carcinoma model was developed by BPS (biomarker pattern software). Results Compared with the healthy people group, there were 34 significant different protein peaks in the gas- tric carcinoma group. And the diagnosis model composed with 6 proteins (M/Z values were 6016, 6744, 2822, 4474, 7892 and 3242) could classify the 2 groups correctly. In the test group, the sensitivity and specificity were 96% and 93.75% respectively, the accuracy was 95.45%. Conclusion SELDI-TOF-MS method show features such as microcontent, fast and high- resolution etc. It could be utilized to screen significant proteins and develop an auxiliary diagnosis model in gastric carcinoma, which may be used to diagnose this disease.

BACKGROUND:In treatment of lumbar diseases, lumbar fusion therapy fails in 20%of cases and may lead to a series of complications such as pain, intervertebral space col apse, and delayed kyphosis deformity.
OBJECTIVE:To observe the effect of nucleus pulposus on interbody fusion after removing the anterior column disc of rabbit lumbar vertebra.
METHODS:A total of 36 healthy Japanese white rabbits were randomly and equal y divided into three groups, with 12 rabbits in each group. (1) Group of anterior longitudinal ligament+bone grafting:The L3 intercalated disc were wel exposed and anterior longitudinal ligament was stripped, obtaining a space to L3 intercalated disc, then the iliac bone was implanted. (2) Group of excising anterior 1/3 disc+bone grafting:After the anterior 1/3 disc tissue was excised, the iliac bone was implanted and sutured as Group of anterior longitudinal ligament+bone grafting. (3) Group of excising anterior 1/3 disc+fixation:After the anterior 1/3 disc tissue was excised, the iliac bone was implanted and the anterior column fixation was performed.
RESULTS AND CONCLUSION:Biomechanical testing showed that, at 12 weeks, the verticality tensile force in the group of anterior longitudinal ligament+bone grafting was obviously higher than other two groups, and have better fusion rate and could bear stronger force. Lateral position lumbar radiography showed that, the bone graft was absorbed and no new bone grew into the intervertebral space in the group of excising anterior 1/3 disc+bone grafting at 12 weeks;the formation of osseous bridge was found in the group of excising anterior 1/3 disc+fixation;complete bony fusion was found in the group of anterior longitudinal ligament+bone grafting. Histological examinations showed that, at 12 weeks, no bone tissue formed in the group of excising anterior 1/3 disc+bone grafting;a smal amount of bone trabecula and osteocytes were observed in the group of excising anterior 1/3 disc+fixation;a great quantity of newborn bone trabecula and osteocytes, remodeling lamel ar bone and canalis haversi structure were observed in the group of anterior longitudinal ligament+bone grafting. The stability of anterior column has notable effect on interbody fusion, after the anterior column disc is destroyed, the free nucleus pulposus may affect spinal fusion, so restoring the stability of the anterior column may promote interbody fusion, but stil cannot get solid spinal fusion.

Animals ; Antibiotics, Antineoplastic ; administration & dosage ; toxicity ; Apoptosis ; drug effects ; Daunorubicin ; administration & dosage ; toxicity ; Disease Models, Animal ; Female ; In Situ Nick-End Labeling ; Male ; Myocardium ; cytology ; metabolism ; pathology ; ultrastructure ; Neuregulins ; metabolism ; pharmacology ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Receptor, ErbB-2 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction