AIM: To observe the expression of COX-2 in rat corneal neovascularization (CNV) and its relationship to CNV, and to explore the inhibition of Celecoxib, a COX-2 inhibitor, to CNV.METHODS: The distribution and quantification of COX-2and VEGF was detected by immunohistochemistry. Expression of COX-2 and VEGF mRNA was quantified by RT-PCR.The difference in protein and mRNA expressions of COX-2and VEGF was analyzed to find the correlation between them.RESULTS: Expression of activated COX-2 and VEGF protein and mRNA in CNV had a dynamic change. VEGF and COX-2co-localized. Compared with the control group, expression of both protein, mRNA of COX-2 and VEGF in experimental group Ⅱ and Ⅲ had significant difference (P＜0.05), indicating the correlation between COX-2 and VEGF, while that in experimental group I had no statistical difference (P＞0.05).CONCLUSION: COX-2 expression was up-regulated in inflammatory CNV. COX-2 modulates the expression of VEGF,playing a very important role in CNV. Celecoxib inhibit COX-2expression so as to hold back the CNV.

Background Recurrence of pterygium is a common complication after the surgical excision of pterygium,and this procedure is related to cell proliferation,inflammation and neovascularization.Researches determined that rosiglitazone can suppress inflammation and neovaseularization and inhibit proliferation,hut few studies concerning the effect of rosiglitazone on pterygium were performed. Objective The aim of this study was to investigate the effect of peroxisome proliferator-activated receptor γ agonist on the proliferation and apoptosis of human pterygium fibroblasts(HPFs)in culture and search for a new drug to prevent and cure the recurrence after pterygium surgery. Methods Human pterygium samples were obtained during surgery and HPFs were cultured and purified using an explant method and 0.25%trypsin digestion,respectively.The identity of cultured HPFs was confirmed by immunohistochemistry using anti-vimentin and keratin antibodies.Rosiglitazone with the concentrations of 0(control),5,10,25,50,75,100,150,200,400μmol/L was then added in the culture medium for 12,24 or 72 hours.1%DMSO was used as blank control.The MTT method was used to assay the biologic effects of rosiglitazone on HPFs.Cell cycle distribution and apoptosis of HPFs after rosiglitazone treatment were studied by flow cytometic analysis.The expression of proliferating cell nuclear antigen(PCNA)mRNA in HPFs was detected by real-time PCR. Result Cultured HPFs radially migrated outward from the pterygium block,and then grew in long fusiform shape,showing positive response for vimentin and negative for keratin.The HPFs became round and thin with loose distribution after the addition of rosiglitazone.Following 25-125 μmol/L rosiglitazone administration for 12,48 or 72 hours,the A490 value of HPFs significantly declined with the increase of dosage(F=158.312,P=0.006)and lapse of time(F=1.924,P=0.135).Following the treatment of 25,75 or 125 μmol/L rosiglitazone for 24 hours,the number of HPFs in G0/G1 phase was markedly elevated;while the cell numbers in S phase decreased significantly in comparison with the control group(P＜0.05).The apoptotic rate of HPFs in the 25,75 and 125 μmol/L rosiglitazone groups significantly increased with the increase of rosiglitazone concentration(P＜0.05).Real-time PCR revealed that after 24 hours of rosiglitazone treatment,the expression of PCNA mRNA in HPFs was suppressed in a dose-dependent manner(F=3244.329,P＜0.05). Conclusion Rosiglitazone inhibits HPFs proliferation,arrests their cell cycle progression in G0/G1 phase,induces apoptosis of HPFs and depresses the synthesis of PCNA in a dose-and time-dependent manner.

Animals ; Hypertension ; metabolism ; physiopathology ; Male ; Myocardium ; metabolism ; Peptidyl-Dipeptidase A ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Receptors, G-Protein-Coupled ; genetics ; metabolism

Objective To investigate the effect of nesfatin-1 (NSF-1) on T-type Ca2+ channel currents in adult mouse dorsal root ganglion (DRG) neurons and possible signal transduction mechanisms involved.Methods We measured the expression of melanocortin 4 receptors(MC4-R)in mouse DRG by using western blotting analysis.The whole-cell patch clamp technique was used to record the effects of NSF 1 on T-type Ca2+ channel currents in small DRG neurons and several ligands were experimented to further clarify relevant signaling pathways.Results MC4-Rs were abundantly expressed in DRG neurons.NSF-1 enhanced T-type calcium channel currents in a dose-dependent manner in small DRG neurons in mice.NSF-1 mediated increment of T-type calcium channel currents was blocked by the MC4-R antagonist HS024,phosphokinase C antagonists GF109203X,and chelerythrine chloride,while the blockade of phospohokinase A PKI 6-22 elicited no such effects.Conclusions NSF-1 can enhance T type calcium channel currents in small DRG neurons through an MC4-R-dependent PKC signaling pathway.

Objective To observe the serum levels of homocysteine (Hcy) and hypersensitivity C-reactive protein(hs-CRP) in elderly patients with cerebral infarction and to investigate their relationship and clinical significance by National Institutes of Health Stroke Scale (NIHSS).Methods The serum levels of Hcy and hs-CRP were by enzymatic cycling method and scattering turbidimetry in the elderly patients ( 116 cases with cerebral infarction and 100 cases of healthy control).Those 116 cases with cerebral infarction were divided into three groups by the degree of NIHSS.The three groups were compared with each other.Results The levels of serum Hcy and hs-CRP in elderly patients with cerebral infarction were significantly higher than that of healthy control group ( t =6.97,P ＜0.01 ; t =14.96,P ＜0.01 ).There has significant difference among those three groups with cerebral infarction by comparing with each other( F =23.49,P ＜0.05; F =28.19,P ＜0.05).A positive correlation was found between Hcy and degree of NIHSS( r=0.54,P ＜0.05),and between hs-CRPand degree of NIHSS( r =0.58,P ＜0.05).Conclusions Serum levels of Hcy and hs-CRP are correlated with the occurrence of cerebral infarction and its severity.There has positive clinical significance to evaluate the effect of cerebral infarction by measuring the serum levels of Hcy and hs-CRP dynamic.

Objective To investigate radiation-induced alternations of phospholipids in epithelial cells,and to provide experimental evidence for understanding the mechanism of radiation-induced intestinal injury.Methods The intestinal epithelial cells(IEC-6)in rats were divided into three groups:normal control group,8 Gy X-ray irradiation group and 12 Gy X-ray irradiation group.Phospholipids were extracted at 6 h or 24 h after radiation and then measured by high-performance liquid chromatography and mass spectrometry(HPLC-MS).Results At 6 h after radiation,the phospholipids in 8 Gy irradiation group didn't vary significantly,while those in 12 Gy irradiation group changed.The PG,PI and Lyso PC were significantly up-regulated(F=5.37,9.60,9.88,P＜0.05).However,at 24 h after radiation,many PE and PG species in both irradiation groups declined(F=5.15-99.77,P＜0.05)and SM species increased in 12 Gy irradiation group(F=4.35-7.92,P＜0.05).Conclusions The ionizing radiation could disorder phospholipid metabolism in IEC-6 cells with a dose-dependent manner.

Objective To study the relationship between morning blood pressure surge with carotid artery intima-media thickness in good controlled elderly hypertensive patients.Methods A total of 151well controlled elderly hypertensive patients was selected in this study.Through the ABPM examination,the morning blood pressure peak was calculated,and then these patients were divided into two groups according to the morning blood pressure peak.The patients whose morning blood pressure peak ≤30 mmHg were divided into non-morning blood pressure surge group (NMS group),and the patients whose morning blood pressure peak ＞ 30 mmHg were divided into morning blood pressure surge group (MS group).The carotid IMT of these patients was measured with ultrasonic detection.The hypertension-related factors with blood pressure and morning peak phenomenon and the impact of IMT were analyzed,and the relationship between the morning blood pressure peak and IMT was analyzed with linear regression analysis.Results Two groups of 151 cases were well-controlled hypertension,76 patients with morning blood pressure showed peak phenomenon,accounting for 50.3％.Age,gender,body mass index,blood lipids,blood glucose,the maximum systolic blood pressure,average systolic blood pressure,minimum systolic blood pressure,maximum diastolic blood pressure,average diastolic blood pressure and minimum diastolic blood pressure had no difference between the two groups ( P ＞ 0.05 ).However,the morning blood pressure peak in patients with MS group [ (42.34 ± 7.10)mmHg] and IMT [ (0.89 ± 0.13 )mm] was higher than the NMS group [ (21.16 ±5.23) mmHg,(0.84 ±0.14) mm,P ＜0.01 orP ＜0.05],and carotid IMT and peak morning blood pressure was positively correlated ( r =0.56,P ＜0.01 ).Conclusions Good controlled elderly hypertensive patients remained the phenomenon of the morning blood pressure surge,the morning blood pressure peak might lead to carotid atherosclerosis.

Objective:To develop HMGA1-silencing SMMC-7721 cell and investigate its impacts on apoptosis and cell cycle in tumor cells.Methods:Constructing HMGA1-silencing SMMC-7721 cell through G418 selection of RNAi plasmid transfected cells;surveying the apoptosis,prliferation and cell cycle of the tumor cells by FACS and MTT.Results:Stable HMGA1-silencing cells were obtained successfully.The apoptosis percentage in RNAi group(29.46?3.04%) was significantly higher than that in the negative control (1.96?0.76%) or control (2.04?0.70%)within each P

Objective:To establish a mouse fibroblastic cell line stably transfected with HMGA1 gene,and to use the cell line to investigate the role of HMGA1 in tumor development and progression.Methods:Eukaryotic expression vector pcDNA3.0-HMGA1 was transfected into NIH3T3 by lipofectamine-mediation.Stable transfectants were selected by G418.The expression of extraneous HMGA1 gene was analyzed by RT-PCR and DNA sequencing.Cell growth was measured through MTT and the cell cycle by FCM.Soft agar colony formation was employed to analyze the ability of anchorage-independent growth.The expression of the immune inhibition factors of VEGF and FasL mRNA was detected by RT-PCR.Results:NIH3T3 cell lines stably expressing HMGA1 gene were successfully established.Comparing with controls,the HMGA1 gene-transfected cells grew faster,acquired the ability of anchorage-independent growth and expressed the immune inhibition factors of VEGF and FasL mRNA.Conclusion:Extraneous expression of HMGA1 gene can induce malignant transformation of NIH3T3 and modulate mRNA expression of immune inhibition factors.HMGA1 gene plays a key role in tumor development,progression and immune escape.

Aim: To compare the stability of Endostar~(TM) and endostatin under different temperatures and pH using polyacrylamide gel electrophoresis( PAGE) and Western blot and to compare the activity of Endostar kept at 4 ℃ and 37 ℃ by inhibition of endothelial cell proliferation. Methods: Endostar and endostatin expressed by Phicha pastoris were kept at 4 ℃, and 37 ℃ for 96 hours, respectively. The electrophoresis of the samples was detected by reduced and non-reduced PAGE. The results were further confirmed by Western blot with rabbit anti-Endostar polyclonal antibody. The inhibitory effect of Endostar stored at different temperatures on HUVEC was also exam-ined by cell-based assay. Results: Single band at 20 kD was detected in all lanes of SDS-PAGE gel loaded with endostatin and Endostar samples under reducing condition. In acidic native PAGE with three different pH values, endostatin showed a smear characteristic, whereas Endostar showed a unique band in acidic non-continuous native PAGE. Although the smear phenomenon was also observed under two conditions of constant native elec-trophoresis, the major band of Endostar could be detected. Similar electrophoretic behavior was found for endosta-tin and Endostar stored at both 37 ℃ and 4 ℃ . Western blot showed similar results to those by PAGE. Furthermore, Endostar stored at these two temperatures also had identical inhibitory effect on proliferation of HUVEC. Conclusion: Endostar and endostatin exhibit similar thermostability at regular conditions, but Endostar was more stable than endostatin expressed in P. pastoris under acidic condition.