Objective To determine the role of CTGF in PVR, we successfully down - regulated the expression of CTGF by RNA interference( RNAi) technology in ARPE-19 cell line. Methods Design and synthesis three CTGF siRNA, screen effective targeted points in Western blot, combine Oligo DNA of effective targeted point, anneal to form double chain DNA, connect with linearing pGC-LV-GFP carrier, sequence DNA in PCR screening positive clones. Results The shRNA-expression vector of CTGF was constructed successfully. And RT-PCR and Western Blot results showed that CTGF expression was significantly inhibited by siRNA transfectants in ARPE-19 cells at mRNA and protein levels. Conclusion We successfully construct shRNA-expression vector of CTGF which paves a way for CTGF-targeted gene therapy of proliferative vitreoretinopathy.

Objective: To construct recombinant vectors for RNA interference(RNAi) targeting Pygo2, and to assess its influence on the proliferation, invasion of glioblastoma U251 cells and the related mechanism. Methods: A pair of oligonucleotides containing short hairpin structure targeting Pygo2 cDNA sequences were designed and synthesized, and their negative control sequences were also synthesized. After annealed, they were inserted into pSuper vector to generate the recombinant plasmids. Then the recombinant plasmids were digested with EcoR I and Hind III for identification, and the sequence was assayed by DNA sequencing. The recombinant plasmids were transfected into cultured glioblastoma U251 cells using Lipofectamine™ 2000. The effect of Pygo2 shRNA on Pygo2 mRNA and protein in U251 cells was detected by real-time PCR and Western blotting analysis, respectively. MTT assay was used to detect the cell proliferation; cell cycle was analyzed by flow cytometry; Bromodeoxyuridine (BrdU) incorporation analysis was used to examine DNA synthesis; and cell invasion assay was performed using Transwell chambers. The effect of Pygo2 shRNA on the protein level and subcellular location of cyclin D1 and β-catenin was detected by Western blotting analysis and immunofluorescent staining. Results: The recombinant plasmids were completely coincided with the design by the restriction map and the sequence analysis. Pygo2 mRNA and protein expression was significantly suppressed by Pygo2 shRNA. Furthermore, the proliferation of cells in Pygo2 shRNA group was notably inhibited, cell cycle was arrested at the G1 phase, and BrdU incorporation and migrating cells were significantly inhibited. In addition, Pygo2 knockdown significantly down-regulated cyclin D1 expression without altering the subcellular location, and the expression level and subcellular location of β-catenin had no noticeable changes. Conclusion: The recombinant vectors for specific suppression of Pygo2 expression have been constructed successfully. Inhibition of Pygo2 expression can suppress cell proliferation and invasion of glioma U251 cells, decrease DNA synthesis, arrest cell cycle at the G1 phase, and decrease expression of the Wnt target gene cyclin Dl.

Female ; Humans ; Hyperthyroidism ; drug therapy ; etiology ; Infant, Newborn ; Male

Objective To investigate the effects of desflurane and/or nitric oxide (NO) on the lung injury induced by ischemia-reperfusion ( I/R) during cardiopulmonary bypass (CPB) in children with pulmonary hypertension secondary to congenital heart disease (CHD) .Methods Forty children with CHD and pulmonary hypertension (24 male, 16 female) aged 0.6-3.7 yrs weighing 7.1-11.9 kg undergoing cardiac surgery under CPB were randomly divided into 4 groups ( n = 10 each): group control; group DES; group NO and group DES + NO. The patients were premedicated with oral midazolam 0.5 mg?kg-1. Anesthesia was induced and maintained with fentanyl and vecuronium. Radial artery was cannulated for MAP monitoring and blood sampling. Pulmonary catheter was placed under direct vision after chest was opened. The patients received inhalation of desflurane (1-1.3 MAC) (group DES)/NO (10-20 ppm) (group NO) /DES + NO (group DES + NO) immediately after pulmonary catheterization until the start of CPB. MAP, PAP, peak airway pressure (Ppeak) and compliance of respiratory system (Crs) were recorded at 5 min after induction (T0 ), 5 min before CPB (T1 ), 5 min after start of CPB (T2) and at the end of surgery (T3) . Blood samples were taken at T0 and T3 for determination of methemoglobin (Met-Hb), soluble intercellular adhesion molecule-1 (sICAM-1), XOD and MDA.Results The four groups were comparable with respect to age, M/F ratio, body weight, duration of CPB and type of operation performed. MAP was significantly decreased after inhalation of desflurane alone or desflurane + NO before CPB ( at T1) as compared to the baseline value at T0; while PAP was significantly decreased after inhalation of NO alone or NO+ desflurane before CPB (at T1) as compared to the baseline value. Ppeak was significantly lower while Crs was significantly higher at end of surgery (T3) in group NO and NO + desflurane than in control group. Blood sICAM-1 and MDA concentrations and XOD activity were significantly lower at the end of surgery (T3 ) in group DES, NO and DES + NO than in control group. Met-Hb was significantly increased at the end of surgery (T2) as compared to the baseline (T0) in group NO and DES + NO but was still within normal range. Conclusion Inhalation of desflurane and/or NO can ameliorate the lung injury during CPB in children with pulmonary hypertension secondary to CHD, in addition to inhibit the pulmonary hypertension.

Objective To study the mechanism of Danshi Qing Tablet (DQT) in treating cho lelithiasis. Methods Sixty guinea pigs were randomly allocated to five groups : Group A (normal control), Group B (model control), Group C (treated with low d osage of DQT), Group D (treated with high dosage of DQT) and Group E (treated w ith Xiaorong Gandanjieshi Tablet). Guinea pig models of cholelithiasis were esta blished by subcutaneous injection of lincomycin. Results After six weeks of treatment, the content of the bile acid (BA) was higher and the content of tota l bil irubin TBIL and Ca 2+ was lower in Group C and Group D than those in Group B. The ga llstone formation rate in Group D was 27.27%, which was lower t han that in Group B (80%), the difference being significant (P

Background Flickering light is different from the normal light environment.Animal experiment proved that flickering light can induce myopia.But its mechanism remains unclear.Objective This study was to investigate the expression of c-fos gene in retina of myopic C57BL/6J mice induced by flickering light and monocular form deprivation.Methods Ninety clean C57BL/6J mice aged 28-day-old with the similar refraction in both eyes were randomly assigned to five groups.Fifteen mice in the control group were exposed to continuous white light environment.The white flickering light with the frequency of 10,5,2 Hz were used to irradiate the mice respectively in high frequency flickering group (15 mice),moderate frequency flickering group (15 mice) and low frequency flickering group (15 mice),respectively.The right eyes of other 30 mice were monocularly occluded with a semitransparent hemispherical thin plastic shell to establish the form deprivation models and then were exposed to white light environment.The diopter and ocular axial length were measured by murine-specific eccentric infrared photorefraction and A-scan ultrasonography before experiment and two weeks after the treatments.At the end of experiment,the mice were sacrificed by neck dislocation.Mice eyes were enucleated and retinal samples were prepared for the detect of c-fos protein and its mRNA by immunohistochemistry,Western blot and reverse transcription polymerase chain reaction (RT-PCR),respectively.Results Immunohistochemistry showed that the expressing rate ofc-fos protein in retina was (68.000±10.368)％,(51.000±6.519)％,(46.000±6.519)％,(31.000±7.416)％ and (25.000 ± 7.071)％ in the control group,high frequency flickering group,moderate frequency flickering group,low frequency flickering group and form deprivation group respectively 2 weeks after experiment.The expression rates of c-fos protein in retina in different frequencies of flickering light groups and form deprivation group were significantly lower than that in the control group (t =3.104,4.017,6.490,7.661,all P＜0.05),with the lowest rate in the form deprivation group (P＜0.05).The expression of c-fos detected by Western blot assay exhibited that the relative values of c-fos protein in retina (c-fos/GAPDH) was 0.804±0.050,0.687±0.047,0.667±0.036,0.558±0.036 and 0.532 ±0.056,respectively in the control group,high frequency flickering group,moderate frequency flickering group,low frequency flickering group and form deprivation group,illustrating significantly lowing in different frequencies of flickering light groups and form deprivation group compared with control group (t =2.961,3.184,6.971,6.276,all P＜0.05),whereas the c-fos in the low frequency group and form deprivation group,c-fos protein was less expressed in comparison with the higher frequency flicking group (P＜0.05).The expression level of c-fos mRNA (c-fos mRNA/GAPDH mRNA) in retina was 0.820±0.056,0.663±0.061,0.627±0.034,0.521±0.041 and 0.474 ±0.045 in the control group,high frequency flickering group,moderate frequency flickering group,low frequency flickering group and form deprivation group,respectively.These results demonstrated a significant decline in the expression of c-fos mRNA in different frequencies of flickering group and form deprivation group compared with the control group(t=3.262,5.070,7.173,8.305,all P＜0.05),and the inhibition ability of low frequency of flickering group and form deprivation group was much stronger.Conclusions The c-fos gene level in the retina has a negative relationship with the severity of myopia induced by flickering light and form deprivation.

Objective To evaluate the influence of different field-strength magnets(1.5 T and 3.0 T)on MRI artifacts caused by Ni-Cr alloy fixed prostheses.Methods The crown,bridge and upper denture fixed prostheses with different thickness were produced by Ni-Cr alloy as test samples,and were one by one put on the centre of water phantom for MR scanning with different field-strength magnets(1.5 T and 3.0 T).The artifact areas on these two field-strength magnets were measured and statistically compared.The plastic prostheses with the same shape and thickness as the test samples were served as controls.Results Ni-Cr alloy fixed prostheses could cause MRI artifacts,and the artifact areas increased with the mass of prostheses.However,no artifact area was found in controls.Compared with those on 1.5 T magnet,the MRI artifact areas significantly increased on 3.0 T magnet(P

By using the diagnosis related groups(DRGs) method, we analyze and compare the rationality and feasibility of outcomes and human resources distribution of clinical specialties and health care professionals.It may increase the hospital management levels and feasibility.The introduction of DRGs for the majority of hospital management may enhance the performance evaluation and human resource levels in the hospitals.

Objective To investigate the application value of multi-slice CT enterography (MSCTE)in diagnosing intestinal tuberculosis (ITB)and Crohn’s disease (CD).Methods MSCTE findings were retrospectively analyzed in 25 patients with ITB and in other 25 patients with CD diagnosed through endoscopy,pathologic examination and clinical follow-up.Statistical analysis was performed to find out the difference in CT findings between the ITB and CD.Results 25 patients with CD included the involved ileum in 23,involved duode-num and jejunum in 8,multiple segmental lesions in 20,asymmetrically thickened intestinal wall in 20,hierarchical reinforcement in 1 9,mesenteric vascular hyperplasia in 20,fibrofatty hyperplasia in 18,peritoneal abscess or fistula in 8,anal fistula in 1,and pseu-do-diverticulum formation in the intestinal wall on the opposite side of the mesentery in 2.Meanwhile,the other 25 patients with ITB included the involved terminal ileum in 25,symmetrically thickened intestinal wall in 23,annular enhancement of lymph nodes in 11,thickened peritoneum and omentum together with distinct enhancement,or intestinal adhesion,or ascites appeared in 15.The CD was more likely to represent multiple segmental lesions,asymmetrically thickened intestinal wall,hierarchical reinforcement,mesenteric vascular hyperplasia,fibrofatty hyperplasia,and peritoneal abscess or fistula formation (P <0.05).Meanwhile,the ITB was more likely to represent the lesion only in ileum,symmetrically thickened intestinal wall,and annular enhancement of lymph nodes (P <0.05).Conclusion MSCTE shows promising clinical application in diagnosis and differential diagnosis of CD and ITB.

objective To explore the role of cytokines and dendritic cells (DCs) in rat high-risk corneal allograft survival pro- longed by superantigen Staphylococcal Enterotoxin B (SEB) and to compare the different effects between SEB and glucocorticoid. Design Experimental study.Participants Fisher 344 and Lewis rats.Methods The Fisher 344 and Lewis inbred rats were used for high-risk penetrating keratoplasty model.All of the Lewis rat recipients were divided into three groups by blinded fashion.GroupⅠand GroupⅡrats were injected intraperitoneally with 0.2ml saline buffer or SEB (75?g/ml) respectively at 4-day intervals on three occasions before transplantation.GroupⅢrats were injected subconjunctivally with 0.1ml dexamethasone (1mg/ml) daily from the first day after surgery for 2 weeks.The allograft survival was examined under slit-lamp.The concentration of interleukin IL-1?,IL-2,IL-4,IL-5,IL-6, IL-10,TNF-?in rat aqueous humor and peripheral blood were measured by liquichip and the cytokine and CD11c,CD80,MHC-Ⅱex- pression in corneal grafts were examined by immunohistochemestry staining.Main Octcome Measures Mean survival time of the allo- grafts,the level of cytokines in aqueous humor,peripheral blood and corneal grafts.Results Compared with group control,the grafts mean survival time was delayed about 3.8d in group SEB (P=0.00) and 7.1d in group dexamethasone(P=0.01).But there was no signifi- cant difference between group SEB and dexamethone (P=0.26).Liquichip test showed that the level of IL-1?in aqueous humor was re- duced and IL-4,IL-6 and IFN-?,ascended.Only IFN-?,and IL-6 could be found in peripheral blood,and the changing shift of them was similar to that in aqueous humor.The immunohistochemistry staining showed that the expression of IL-2 in rat corneal grafts was signif- icantly decreased but IL-4,IL-6 and IL-10 elevated.The expression of DCs in group SEB was similar to that in group control,which was elevated after keratoplasty,but the phenotype of DCs was not the same.There were predominantly mature DCs in group control while immature DCs in group SEB.Conclusions The immunological inhibiting effect of SEB is same to glucocorticoid,but the mecha- nism is different.SEB can modulate immune response,which might induce immune inhibition via the local production of cytokines and effect on DCs maturation involving corneal graft rejection prevention.