Neovascular age-related macular degeneration (AMD) is the main cause of vision loss in AMD patient.Now,the application of anti-vascular endothelial growth factor(VEGF) has become the preferred therapy for the treatment of neovascular AMD,and it has been proved to improve the visual acuity and reduce the blindness rate.But the formation of choroidal neovascularization (CNV) in AMD is a comprehensive and integrated pathological process.It is difficult to cure the CNV with only anti-VEGF drug for all the neovascular AMD patients.Therefore,the targeting drugs toward other signal pathway associated with CNV are being in different phases of clinical trials.Resent years,with the development of science and technology,small interference RNA(siRNA) and genome technology has been applied to treat neovascular AMD and show a potential prospect,and gene therapy and stem cell therapy are the most remarkable and predominant ways for AMD.Doubtlessly,these therapies offer novel choice for the management of neovascular AMD.

Objective To establish a newborn animal model of renal injury caused by intrauterine asphyxia and explore the mechanism of renal injury in neonate after asphyxia.Methods After two-horn uterus and vessels supplying uterus and ovary were exposed in 21-day-pregnant Wistar rats,arterial clamp occluded one side of vessels.The occluding time were 10 and 30 minutes.Then arterial clamp was taken off,and reperfusion for 30 minutes,2,6,12 and 24 hours respectively.Reaching prescribed time uterus horn was opened rapidly and pups were removed.The pups sacrificed by decapitation.Kidneys were taken out and studied by HE staining and electron microscope.Results Kidney of fetal rats in 21 gestational age was developmental and mature degree of tubules dropped behind that of glomerule.Changes of proximal tubules were early and serious compared with distal tubules during ischemia and reperfusion stages.Conclusion Ischemia and reperfusion to graded pregnant rat can supply an ideal model to study injury of kidney and other organs(intraute)-rously.

Objective To investigate the alterations of serological endothelin(ET) in patients with kawasaki disease(KD) and its relation with coronary dilatation(CAD).Methods Serological ET were measured in 50 cases of patients with KD in acute phase as well as subacute phase; 30 cases of patients with acute febrile infection(IC) in infective phases and 30 healthy children(HC).Results 1.ET in subacute phase significantly increased than that in acute phase.2.In both acute and subacute phases of KD, ET significantly elevated higher than that in HC; and ET in subacute phase significantly elevated than that of IC. 3. Fifty-four percentage patients with KD were complicated with CAD. For CAD subgroup, ET had no difference with CAD subgroup in subacute phase.Conclusion ET has still increases in subacute of KD,which indicates the relation with coronary artery lesion.

OBJECTIVETo study the cleavage activity on the HCV RNA of a chimeric recombinant of HCV specific ribozyme and U1 small nuclear RNA, which compartmentalizes within the nucleolus.

METHODSThe third stem-loop sequence of human U1 snRNA (position 95-116) within pBSIISK+ U1 was substituted by hammerhead ribozyme against HCV RNA by PCR and cloning methods, and the constructed plasmid was named pBSIISK+ (U1-Rz). Then the whole gene fragment of the chimeric ribozyme was cloned into a pGEM-T vector under the control of T7 promoter, and the constructed plasmid was named pGEM- (U1-Rz). The pGEM- (U1-Rz) and pGEM-Rz (containing the same ribozyme sequence as that in U1-Rz) transcripts as enzyme were transcribed in vitro. Also the (32)P-labeled pCMV/T7-NCRC luc (containing the gene sequence of the whole 5'-NCR and part core of HCV RNA) transcripts as target-RNAs were transcribed in vitro. The enzymes were incubated with the target RNAs under different conditions and autoradiographed after denaturing gel-electrophoresis.

RESULTSThe sequencing result showed that the construction of U1 snRNA chimeric ribozyme was correct. Compared with the ribozyme alone, both of them were active at 37 degree C and with Mg2+ (10 mmol/L) and TrisCl (10 mmol/L, pH7.9), and there was no remarkable difference between them. The cleavage activity of the chimeric ribozyme increased with the prolongation of reaction time and increment of enzyme concentration.

CONCLUSIONBoth ribozyme and U1 snRNA chimeric ribozyme exhibited specifically catalytic activity against HCV RNA in vitro. There was no remarkable difference between their cleavage efficiencies.

Chimera ; genetics ; Genetic Therapy ; Hepacivirus ; genetics ; Hepatitis C ; therapy ; RNA, Catalytic ; genetics ; metabolism ; RNA, Small Nuclear ; genetics ; pharmacology ; RNA, Viral ; genetics ; Recombinant Fusion Proteins ; pharmacology

To determine the optimum conditions of supercritical CO2 extraction of Xiaoyaosan, and establish its fingerprint by gas chromatography-mass spectrometry (GC-MS), the yield of extract were investigated, an orthogonal test was used to quantify the effects of extraction temperature, pressure, CO2 flow rate and time, and fingerprint analysis of different batches of extracts were by GC-MS. The optimal extraction conditions were determined as follows: extraction pressure 20 MPa, extraction temperature 50 degrees C, CO2 flow rate 25 kg x h(-1), extraction time 3 h, and average yield 2.2%. The GC-MS fingerprint was established and 27 common peaks were found, whose contents add up to 81.89% of the total peak area. Among them, 21 compounds were identified, accounting for 53.20% of the total extract. The extraction process is reasonable and favorable for industrial production. The GC-MS method is accurate, reliable, reproducible, and can be used for quality control of supercritical CO2 extract from Xiaoyaosan.
Carbon Dioxide ; chemistry ; Chromatography, Supercritical Fluid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Gas Chromatography-Mass Spectrometry ; methods

OBJECTIVETo compare alveolar bone mineral density (BMD) changes of the male and female diabetes rats, and male diabetic rats BMD in systemic change.

METHODS40 Wistar rats were divided into diabetic group (15 female rats and 15 male rats) and control group (5 female rats and 5 male rats). Models of diabetes rats were made by peritoneal injection of streptozotocin (STZ). Blood glucose, serum insulin, indicators of bone metabolism as well as BMD of mandibular alveolar bone, femur and lumbar spine were examined.

RESULTSThe BMD of mandibular alveolar bone in male rats was higher than the female rats in the two groups (P<0.05). The BMD of mandibular alveolar bone of male and female diabetic rats in the diabetic group was lower than that in the control group (P<0.05). The BMD of male rats in the control group was femur, mandibular alveolar bone, lumbar spine from high to low. The BMD of male rats in the diabetic group was mandibular alveolar bone, femur, lumbar spine from high to low.

CONCLUSIONOsteoporosis happen to mandibular alveolar bone of female and male diabetes rats, especially in female rats. The BMD of mandibular alveolar bone, femur and lumbar spine in the male diabetic rats were lower than that in the control group. Mandibular alveolar bone density changes of diabetic rats was in line with the lumbar spine and femur's, or mandibular alveolar bone density changes of diabetic rats lags behind the lumbar spine and femur's.

Animals ; Bone Density ; Diabetes Mellitus, Experimental ; Female ; Femur ; Lumbar Vertebrae ; Male ; Mandible ; Osteoporosis ; Rats ; Rats, Wistar

Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; metabolism ; Esophageal Neoplasms ; genetics ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Genes, p53 ; Humans ; Loss of Heterozygosity ; Male ; Middle Aged ; Point Mutation ; Tumor Suppressor Protein p53 ; genetics ; metabolism

Humans ; Stents ; Tympanic Membrane ; Tympanic Membrane Perforation ; surgery

OBJECTIVETo explore the effect of Bushen Huoxue Compound (BHC) on lactate dehydrogenase (LDH) leakage, expressions of vascular endothelial growth factor (VEGF) and VEGF mRNA in retinal Muller cells under high glucose condition or advanced glycosylation end products (AGEs) condition by using serum pharmacological method.

METHODSThe retinal Müller cells of 5-7 days post-natal Sprague Dawley (SD) rats were cultured with modified enzyme-digestion method. Purified retinal Muller cells were cultured in normal conditions, high glucose condition (50 mmol/L) or AGEs (50 mg/L and 100 mg/L) conditions, and BHC-containing serum was added to culture medium. The LDH leakage and VEGF expressions were measured by enzyme-linked immunosorbent assay (ELISA). In addition, the relative expression of VEGF mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSCompared with the normal control group, expressions of VEGF and VEGF mRNA were significantly increased in the high glucose group, the low dose AGEs group and the high dose AGEs group (all P < 0.01). The LDH leakage was obviously increased in the high dose AGEs group, when compared with the normal control group and the high glucose group (P < 0.01). The LDH leakage, expressions of VEGF and VEGF mRNA were obviously decreased by BHC-containing serum both in high glucose and AGEs conditions (P < 0.05, P < 0.01). BHC-containing serum had no significant effect on the LDH leakage and expressions of VEGF and VEGF mRNA in normal conditions (P > 0.05).

CONCLUSIONSAGEs intervention could obviously lower the stability of Müller cell membrane. Up-regulated expressions of VEGF and VEGF mRNA in cultured Müller cells could be induced by AGEs or high glucose. BHC-containing serum could stabilize the stability of Müller cell membrane, inhibit the transcription of VEGF mRNA and decrease the protein expression of VEGF, which might be one of important mechanisms for preventing and treating diabetic retinopathy.

Animals ; Cells, Cultured ; Diabetic Retinopathy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Ependymoglial Cells ; Glucose ; L-Lactate Dehydrogenase ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A

Aneurysm, Dissecting ; genetics ; Aortic Aneurysm ; genetics ; Diseases in Twins ; Humans ; Male ; Middle Aged