BackgroundHypoxia and hyperglycemia are the common causes of retinal neovascularization.In these states,H+ accumulates because of the elevated glycolysis and failure of retinal circulation,thus the retinas readily acidified. ObjectiveThe present study was to explore whether retinal acidosis independently regulates the production of vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) and whether the regulation is related to oxidative stress.Methods The retinas from 2-week-old male SD rats were cultured with explant method in DMEM modulated by NaHCO3,and culture retinas were randomly divided into pH 7.2,6.8 and 6.5 groups for 24 hours.In addition,after 24 hours of culture as above described,retinas were washed using PBS two times and then followed by again culture in DMEM with pH 7.2 for another 24 hours.Also,antioxidant was added in different pH values of DMEM for culture as above described.The retinal samples were prepared for histopathological examination.The expressions of VEGF and PEDF proteins and their mRNA in retina tissue were detected by Western blot and fluorescence quantitative polymerase chain reaction (PCR) respectively.Results The retina showed the clear structure and morphology in pH 7.2 group and pH 6.8 group,but retinal vacuoles change was seen in pH 6.5 group after culture for 24 hours.No significant difference was seen in the expressing level of VEGF mRNA in retina between normal group and pH 7.2 group( 112％ ±11％ vs 100％ ±7％ ) (P=0.55),but those in pH 6.8 group and pH 6.5 group were significant increased in comparison with pH 7.2 group( 196％ ±43％ vs 100％ ±7％ ;251％ ±29％vs 100％ ±7％ )( P＜0.05 ).The expressing level of PEDF mRNA in retina in normal group was similar to that of pH7.2 group(86％ ±19％ vs 100％ ±33％) (P=0.64),but that in pH 6.5 group was significantly higher than pH 7.2 group( 230％ ±66％ vs 100％ ±33％ ) ( P＜0.05 ).The resemble results were found in the expressions of VEGF and PEDF protein.After pH reversion,the expressing levels of VEGF mRNA were 100％ ±13％,111％ ±9％,113％ ±9％ in pH 7.2 group,pH 6.8 group and pH 6.5 group respectively without significant difference among them (F=2.51,P=0.16).The expressing levels of PEDF mRNA were 100％ ±13％,110％ ±9％,108％ ±11％in different groups ( F =0.98,P =0.43 ).Under the presence of antioxidant,the expressing level of VEGF mRNA in pH 6.5 group increased in comparison with pH 7.2 group and pH 6.8 group ( P ＜ 0.05 ).The expressing levels of PEDF mRNA were significant different among pH 7.2 group( 100±31 )％,pH 6.8 group( 282±45 )％ and pH 6.5 group(480±117)％ (F=20.73,P=0.00). Conclusions VEGF can be induced by retinal acidification alone,which may be regulated by oxidative stress.Under the retinal acidification,antioxidants promote the expression of PEDF,suggesting that oxidative stress inhibits the production of PEDF.

Acupuncture Therapy ; Administration, Inhalation ; Aged ; Aged, 80 and over ; Cerebral Infarction ; drug therapy ; therapy ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Oxygen Inhalation Therapy

Objective To investigate the relationship between trace elements and rachitis in children.Methods Three hundred and twelve patients with rachitis and 297 healthy children were selected for this study.Blood zinc(Zn),iron(Fe),plasma copper(Cu),calcium(Ca),magnesium(Mg),lead(Pb) and cadmium(Cd) were assayed by atomic absorption spectrophotometer.Results The levels of Zn,Fe,Cu of rachitis in blood were significantly lower than those of healthy children,while the levels of Mg,Pb were higher.There were significant differences between 2 groups(P

Objective To explore the effect of ca se-teaching applied to the classes of practical training of parasitology. Methods The case-teaching was tentatively performed in a part of the undergraduates in our school during their parasitology practical training classe s.Results The effect of case-teaching was obviously superior t o the traditional one. The total scores,the theory examination and the specimen recognition scores were all higher in the class which employed the case-teachin g than in the control ones (P

OBJECTIVETo construct and identify a lentiviral vector of RNA interference targeting human Notch-1 gene.

METHODSTo determine the Notch-1 gene sequences, three RNAi target sequences (shRNA1-3) were designed in accordance with the RNAi sequence design principles and cloned into the lentiviral vector pLenOR-THM by endonuclease BamH I restriction, EcoR I double digestion, and T4 DNA-ligase ligation. After the transformation into competent DH5alpha bacteria, the candidate clones were identified by Kpn I and EcoR I double digestion and DNA sequencing. The recombinant and three packaging plasmids were co-transfected into human embryonic kidney cell line 293T cells by lipofectamine to produce the lentiviral particles. The viral titer was determined. The 293T cells were infected by the lentiviral particles obtained, and transfection efficiency was assessed using a fluorescent microscope. The lentiviral vector particles were also transfected into ACC-M cells. The Notch-1 expression in the transfected cells was assayed by quantitative reverse transcription polymerase chain reaction (QRT-PCR) and Western blot analysis.

RESULTSThe lentiviral RNAi vector pLenOR-THM-Notchl for Notch-1 gene was constructed successfully. Strong green fluorescence was observed in the 293T cells under fluorescent microscope after co-transfection of the cells with the four plasmids of lentiviral vector. The virus in the supernatant reached a titer of 5.8 x 10(8) TU x mL(-1). The transfection efficiency of the collected virus exceeded 90% in 293T cells with 1 as a multiplicity of infection. The third lentiviral vector was found to significantly inhibit the Notch-1 expression at the mRNA and protein levels.

CONCLUSIONThe lentiviral RNAi vector of Notch-1 has been successfully constructed and identified.

Cell Line ; Genetic Vectors ; Humans ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Receptor, Notch1 ; Transfection

Objective To explore the effect of levels and ratios of matrix nephritis metalloproteinase-2(MMP-2), -9 and inhibitor of metalloproeinase-1(TIMP-1) in the pathogenesis of children with Henoch-Schonlein purpura nephritis(HSPN),and the correlation between them and urinary micro albumin(MA).Methods Serum levels of MMP-2, -9 and TIMP-1 were determined by double antibody enzyme linked immunosorbent assay(ELISA),and urine MA was determined by immune rate nephelometry in 36 children with HSPN,16 children with simple purpura and 30 healthy controls.Results Levels of MMP-2, -9,TIMP-1 and ratios of MMP-2/ TIMP-1,MMP-9/TIMP-1 rose in acute phase of HSPN. The levels and ratios in HSPN group were higher than those in simple purpura group,and those in simple purpura group higher than those in controls (P

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Bone Marrow ; metabolism ; Brain ; metabolism ; Brain Neoplasms ; metabolism ; pathology ; CDC2 Protein Kinase ; metabolism ; Cell Line, Tumor ; Child ; Child, Preschool ; Female ; Gene Expression Regulation, Neoplastic ; Glioma ; classification ; metabolism ; pathology ; Humans ; Male ; Mice ; Mice, Nude ; Middle Aged ; Neoplasm Transplantation ; Neoplastic Stem Cells ; metabolism ; Young Adult

OBJECTIVE: To study the objectivity and reliability of needle electromyography and nerve conduction for detection of musculus extensor digitorum brevis strength, which may provide a basis for establishing a quantitative detection of muscle strength in forensic clinical study. METHODS: Forty-four healthy people were enrolled as the subjects, and during toe dorsiflexion, the following items including needle electromyography indexes, motor unit potential (MUP) amplitude, MUP count, recruitment reaction type, and nerve conduction detection indexes, compound muscle action potential (CMAP) amplitude, CMAP latent period and motor nerve conduction velocity (MNCV), were simultaneously detected under the cooperation and disguise condition. RESULTS: Under the cooperation condition, regardless of the same operator or different operators, there were good test-retest reliabilities in MUP amplitude, CMAP amplitude, CMAP latent period and MNCV, while there were normal test-retest reliabilities in MUP count and recruitment reaction type and the repeatability of the same operator was slightly better than the repeatability between different operators. Under the disguise condition, test-retest reliabilities of MUP amplitude, CMAP amplitude, CMAP latent period and MNCV were relatively high, while test-retest reliabilities of MUP count and recruitment reaction type were relatively low. CONCLUSION There are good test-retest reliabilities in MUP amplitude, CMAP amplitude, CMAP latent period and MNCV, which can be conducive to comparison between different operators and results at various times; MUP count and recruitment reaction type, which can be easily affected by subjectivity of operators and examinees, can be used to differentiate whether an examinee disguises or not. The indexes used to objectively judge muscle strength remain to be further investigated.
Electrodes, Implanted ; Electromyography ; Humans ; Muscle Strength/physiology* ; Muscle, Skeletal/innervation* ; Neural Conduction/physiology* ; Reproducibility of Results ; Toes

Objective To compare prospectively the features and effects of combined operation(splenorenal shunt plus portal-azygous devascularization) and portal-azygous devascularization only(PCDV)on portal(hypertension)(PH).Methods We summarized 360 cases of PH admitted from 1984 to 2004.All patients were randomly divided into two groups,one was combined operative group(250 patients) and the other was PCDV group(110 patients).The therapeutic effects and changes of portal hemodynamics were studied with doppler flowmeter(DCFI),free portal pressure(FPP) and digital subtraction angiography(DSA) pre-and post-operatively,and were measured directly during the course of the procedure.Results(1)Postoperative bleeding:Of all the patients who underwent combined operation,no case of rebleeding occurred in the short period after operation,and the rebleeding rate was 8.0% in the long period of follow-up.In the patients who underwent PCDV,the rebleeding rate was 5.5% in the short period after operation,and 17.6% at long-term follow up(P0.05).(3)There was a significant decrease in the diameter of portal vein,and FPP postoperatively in the combined operation group compared to PCDV group.There was a significant decreases of PVF in the PCDV group.But the decrease of PVF in the two groups had no significant difference.Conclusions The combined procedure has merits of greater decrease of FPP,and alleviation of the condition of hyperdynamic blood flow in the portal vein.The clinical effect is also better than that of portal-azygous devascularization only.

Antigens, CD34 ; metabolism ; Cytokines ; metabolism ; Diagnosis, Differential ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Perineum ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Sarcoma ; metabolism ; pathology ; surgery ; Vimentin ; metabolism