Objective To study the effects of calcium dobesilate plus laser on serum vascular endothelial growth factor ( VEGF ) and insulin-like growth factor-1 ( IGF-1 ) in patients with diabetic retinopathy .Methods 82 patients(93 eyes) were randomly divided into two groups ,the control group(n=41 cases,46 eyes) and the obser-vation group(n=41 cases,47 eyes).The control group was treated through laser ,while the observation group was treated through laser plus calcium dobesilate .Serum VEGF and IGF-1 were measured before and after treatment . Results Before treatment,visual acuity had no significant difference between the two groups (t=0.601,P>0.05). After treatment,visual acuity was improved in 30 eyes in the control group with improving rate of 65.2%,visual acuity improved in 38 eyes in the observation group with improving rate of 80.9%.The improving rate of visual acuity in the observation group was significantly higher than that of the control group (χ2 =6.92,P<0.05).The total effective rate of the control group was 73.9%,which was significantly lower than 87.2%in the observation group (χ2 =9.28,P<0.05). Serum VEGF and IGF-1 had no significant differences before and after treatment in the control group ( t=0.613, 0.496,all P>0.05).After treatment,serum VEGF and IGF-1 in the observation group were (91.4 ±25.1)ng/L and (121.6 ±21.7)mg/L,which were significantly lower than those before treatment (t=3.721,5.992,all P<0.05). Conclusion Calcium dobesilate plus laser can significantly decrease serum VEGF and IGF-1 in patients with diabet-ic retinopathy .

Objective To compare compound staining of Alizarin red S and Alcian blue showing ossified skeletal structures and cartilage in fetal,neonatal and adult rat. Methods All specimens were skinned and fat tissue removed completely and eviscerated,then fixed in 95% ethanol;Acetone was used remove fats; 0.1% Alizarin red S and 0.15%-0.3% Alcian blue were used in ossified bone and cartilage;Differentiation and clearing in 0.5%-2% aqueous KOH. Results Ossified bones showed rose red;cartilage showed blue;the others were transparent.Conclusion The best results of Alizarin red S and Alician blue compound staining could be obtained in all rat skinned,eviscerated,and removed the adipose tissues.

The academic thought of preventing possoble disease is one of the important part of the theoretical system of Chinese medicine. In Pro. WEI Pin-kang's opinion, we must identify the possible diseases prior to preventing it. The article discusses how to identify the possible disease from three aspects, such as indentifying constitution, comprehensive analysis by the four examination methods, drawing assistance from micro-indexex, which to cultivate young physicians' clinical thinking of indentifying possible disease.

Case Method is useful to helping the teacher teach students the knowledge of gynecological diseases and train their capability of clinical thought. The key of the Case Method is choosing the cases and designing teaching steps carefully.

Objective To prepare monoclonal antibodies against VP6 protein of human group A rotavirus(RVA),develop a double-antibody sandwich ELISA detection method,and to verify and preliminarily apply the method,in order to provide more choices for the rapid diagnosis,epidemiological investigation and virus surveillance of RVA in China.Methods VP6protein of human RVA G9P [8] strain was expressed and purified with baculovirus expression system,and then applied to immunize five female BALB/c mice to generate monoclonal antibodies.The median effective concentration(EC_(50)) of the monoclonal antibodies was determined by indirect ELISA,and the binding activity of monoclonal antibodies to RVA was determined by Western blot and indirect immunofluorescence assay(IFA).Using monoclonal antibody pairing,one strain was used as coating antibody and the other strain was labeled with HRP for detection,and a double-antibody sandwich ELISA was developed.The sensitivity,specificity and precision of the method were further verified.RV clinical stool samples were taken and detected by Oxoid ProSpecT commercial kit and the developed double-antibody sandwich ELISA respectively,and the coincidence rate of the detection results was compared.Results The purified soluble VP6 protein was obtained,with a purity of over 90% and an expression level of about 4.5 mg/L.Two strains of RVA VP6 monoclonal antibodies were screened,named 1L3 and 4F22,and the EC_(50) of them binding to VP6 protein was 1.777 and 3.748 ng/mL,respectively,indicating that the two strains of monoclonal antibodies could bind well to RVA.The sandwich ELISA method for detection of human RVA was developed using 1L3 as the capture antibody and HRP labeled 4F22 as the detection antibody.The minimum detectable amount of RVA VP6 protein was 156.25 ng/mL.There was no cross-reaction between the two monoclonal antibodies and human Norovirus(NoV),Sapovirus(SaV) and adenovirus(AdV).The coefficients of variation(CVs) of intra-batch and inter-batch precision verification were both less than 10%.The 192 clinical stool samples of RV were detected by using this method,and compared with the commercial kit results,the positive coincidence rate and negative coincidence rate were 99% and 98%,respectively.Conclusion The double-antibody sandwich ELISA against human RVA developed in this study has good specificity,sensitivity,and precision,which can be used for the detection of human RVA.

Objective To prepare monoclonal antibodies against VP6 protein of human group A rotavirus(RVA),develop a double-antibody sandwich ELISA detection method,and to verify and preliminarily apply the method,in order to provide more choices for the rapid diagnosis,epidemiological investigation and virus surveillance of RVA in China.Methods VP6protein of human RVA G9P [8] strain was expressed and purified with baculovirus expression system,and then applied to immunize five female BALB/c mice to generate monoclonal antibodies.The median effective concentration(EC_(50)) of the monoclonal antibodies was determined by indirect ELISA,and the binding activity of monoclonal antibodies to RVA was determined by Western blot and indirect immunofluorescence assay(IFA).Using monoclonal antibody pairing,one strain was used as coating antibody and the other strain was labeled with HRP for detection,and a double-antibody sandwich ELISA was developed.The sensitivity,specificity and precision of the method were further verified.RV clinical stool samples were taken and detected by Oxoid ProSpecT commercial kit and the developed double-antibody sandwich ELISA respectively,and the coincidence rate of the detection results was compared.Results The purified soluble VP6 protein was obtained,with a purity of over 90% and an expression level of about 4.5 mg/L.Two strains of RVA VP6 monoclonal antibodies were screened,named 1L3 and 4F22,and the EC_(50) of them binding to VP6 protein was 1.777 and 3.748 ng/mL,respectively,indicating that the two strains of monoclonal antibodies could bind well to RVA.The sandwich ELISA method for detection of human RVA was developed using 1L3 as the capture antibody and HRP labeled 4F22 as the detection antibody.The minimum detectable amount of RVA VP6 protein was 156.25 ng/mL.There was no cross-reaction between the two monoclonal antibodies and human Norovirus(NoV),Sapovirus(SaV) and adenovirus(AdV).The coefficients of variation(CVs) of intra-batch and inter-batch precision verification were both less than 10%.The 192 clinical stool samples of RV were detected by using this method,and compared with the commercial kit results,the positive coincidence rate and negative coincidence rate were 99% and 98%,respectively.Conclusion The double-antibody sandwich ELISA against human RVA developed in this study has good specificity,sensitivity,and precision,which can be used for the detection of human RVA.

BackgroundRetinal pigment epithelial (RPE)cell senescence damage the metabolism of photoreceptor,leading to retinal dysfunction and loss of vision.To understand RPE cell senescence mechanism will contribute to the study of age-related macular degeneration ( AMD).ObjectiveThe present study was to prepare the ageing RPE cell model with passage and explore its potential mechanisms.MethodsThis study was approved by the Ethic Committee of Qingdao University Medical College,and the informed consent was obtained from each gravida.Six human eyeballs were obtained from artificial labor fetusl with the gestational age 16-28 weeks.RPE cells were isolated,cultured and passaged in vitro to establish the cell replicative aging model.The third to twelfth cells were collected to be used to this experiment.Human keratin was used to identify the cells by immunochemistry,and MTT method was utilized to assess the proliferation and viability of different generations of cells as the A490 value.The cellular cycles and transmembrane potential (△ψm)of mitochondrion (△ψm) with passage were detected and compared using Flow Cytometry. Results Cultured and passaged cells showed the hexagon in shape with the melanin in 1-2 generations of cells and presented with the brown staining in cytoplasm for human keratin.The melanin was absent in the third generation cells.Vibrant growth statues were seen from the 3-6 generations cells and thereafter the proliferation ability reduced.The cells of G0/G1 phase were gradually elevated with the passage from 3 - 12 generations with a percentage of 68.40％ in the third generation of cells to 87.33％ in the twelfth generation of cells,showing a significant difference among various generations ( F =180.43,P =0.00),and that of the sixth,ninth and twelfth generation of cells was significant higher than the third,sixth and ninth generation respectively (t =4.002,P＜0.05 ; t=12.885,P＜0.01 ;t=16.387,P＜0.01 ).MTT assay showed that of RPE cells were significantly declined with the passage ( F =38.77,P =0.00),and the A490 value of the ninth,twelfth generations of cells was considerably lower in comparison with sixth and ninth generation respectively ( t =5.991,11.983,P＜0.01 ).From 3 through 12 generations of cells,the staining intensity of rhodamine 123 was gradually decreased ( F =121.68,P =0.00 ),and the staining intensity in the sixth,ninth and twelfth generation of cells was significant lower than that the third,sixth and ninth generation respectively(t=6.918,7.620,11.207,P＜0.01 ).Conclusions A replicative aging model can be successfully created by the passage in vitro using human fetal RPE cells.The reduce of transmembrane potential and damage of mitochondria might be one of mechanisms of senescence of RPE cells.

Objective To analyze the diagnose and treatment of drug-induced hypersensitivity syndrome (DIHS) in children.MethodsThe clinical data from one case of highly suspected DIHS were retrospectively analyzed. The related literatures were reviewed.ResultsA 22-month-old male child with severe pneumonia, after treated with vancomycin, suffered with high fever and skin rash combined with hepatic lesion and hematological system disorder. After withdrawal of vancomycin and then treatment with the combination of high-dose methylprednisolone and intravenous immunoglobulin, the clinical symptoms were gradually relieved in 24 hours. The child was improved and discharged on 17th day.ConclusionThe child has the characteristics of DIHS which is highly suspected to be caused by vancomycin.

Objective To evaluate visual field changes in early mild Parkinson's disease.Methods A total of 66 eyes of 33 cases with early mild Parkinson's disease and 72 eyes of 36 age-matched normal individuals were enrolled into the study.Humphrey Field Analyzer Ⅱ was applied for central visual field test.The visual field indices of mean deviation (MD) and pattern standard deviation (PSD) were analyzed to evaluate the location and the characteristics of visual field defect in this study.Results Visual field indices MD (-3.4±2.5) dB was significantly changed in patients with PD when compared to the controls (-0.6 ± 1.7) dB.PSD (4.3±2.6) was significantly higher in patients with PD than that in the control group (2.1 ± 1.8) dB.Glaucoma hemifield test (GHT) assessment was within normal limits in the controls.Of the 33 patients (66 eyes) in PD,GHT showed outside normal limits in 31 eyes,borderline in 8 eyes,and within normal limits in 27 eyes.31 eyes outside normal limits appeared glaucomatous visual field defects,in which 16 with nasal step and 5 with arcuate defect.Conclusions Visual field indices including MD and PSD in early mild patients with PD were significantly worse than that in the controls group.GHT abnormalities could be found in early mild PD patients with visual field defects,including pericentral scotoma and nasal step,which mimicked glaucomatous changes.

Objective The BOLD -fMRI(blood oxygenation level dependent functional magnetic resonance imaging)was used to observe the functional cortical activities of the bilateral cerebral hemispheres in 23 cases of chil_dren with congenital sensorineural hearing loss under resting state ,and to further explore the values of BOLD-fM_RI in patients with sensorineural hearing loss .Methods Twenty three children with congenital sensorineural hearing loss were included in the experimental group and 10 normal heating children were included as cronol group .The ALFF(amplitude of low -frequency fluctuation) method was used to analyze the image data ,and to compare the ex_perimental and control groups with bilateral cerebral cortex function activities for the quantitative analysis .ResuIts The resting state activity intensity in the left inferior temporal gyrus ,left fusiform gyrus ,right cingulate gyrus and left post central gyrus of the experimental group were significantly greater than those of in the corresponding intensi_ty cortex of the control group (P< 0 .01) .ConcIusion The cortex of sensory systems in children with congenital sensorineural deafness may have functional remodeling ,cerebral cortex of patients with sensorineural hearing loss may have perception compensation phenomenon .