Objective To evaluate the role of TCD examination in diagnosis of the early cerebrovascular diseases in aged diabetic patients.Methods 179 cases of aged type Ⅱ diabetic patients were divided into three groups(PDR,BDR and NDB)according to their retinopathy.TCD and retina examination were performed in all patients and the data be analysed. Results (1)The systolic peak flow velocity(Vp) of MCA、ACA、ICA and BA were significant increased in diabetic pa- tients than in normal control group(P

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

In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.
Animals ; Antibodies, Bispecific ; chemistry ; Antibodies, Monoclonal ; chemistry ; Arthritis, Experimental ; Escherichia coli ; Fibronectins ; chemistry ; Half-Life ; Humans ; Mice ; Single-Chain Antibodies ; chemistry ; Tumor Necrosis Factor-alpha ; chemistry

OBJECTIVETo investigate the effect of serum contained Chinese drugs for nourishing Shen and activating blood (S-NSAB) on activity of purified retinal ganglion cells (RGCs) cultured in high glucose medium.

METHODSPurified RGCs of SD rats were cultured in stimulative stable high glucose (50 mmol/L) condition (SHG) and fluctuated glucose condition (FGC) separately, they were intervened with S-NSAB, and the lactate dehydrogenase (LDH) leakage was detected by spectrophotometer for estimating the activity of RGCs.

RESULTSLDH leakage (U/L) in SHG culture was 1 349.17 +/- 215.50 at 24 h, 1220.24 +/- 124.53 at 48 h and 1982.14 +/- 219.03 at 72 h, all significantly lower than that in normal control at the corresponding time points (1628.10 +/-122.10, 1484.13 +/- 127.55 and 2155.75 +/- 140.44, respectively, P<0.05), whereas it was obviously higher in FGC culture at 72 h (2299.60 +/- 88.35), showing that LDH leakage in FGC was significantly higher than that in SHG at the corresponding time points (P<0.05). The LDH leakage was obviously decreased by Chinese medicine intervention with S-NSAB both in SHG at 72 h (1797.62 +/- 146.40) and in FGC at 48 h (1259.92 +/- 87.74) and 72 h (1940.40 +/- 155.47), the difference between pre- and post-intervention was significant (P<0.05).

CONCLUSIONFluctuated glucose conditions of culture medium could obviously damage the membranous stability of RGCs to enhance their permeability and lower the activity of cells; S-NSAB could improve these abnormalities in either SHG or FGC condition, which may be one of the important mechanisms of Chinese formula for nourishing Shen and activating blood in preventing and treating diabetic retinopathy.

Animals ; Animals, Newborn ; Cell Survival ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells ; cytology ; drug effects ; Serum

OBJECTIVETo investigate the impacts of steady high-glucose or fluctuated glucose conditions on glutamate (Glu) release in purified retinal ganglion cells (RGCs) cultured in vitro, and the effect of serum contained Chinese drugs for nourishing Shen and activating blood (S-NSAB) on it.

METHODSRGCs of neonatal SD rats were cultured by antibody combined two-step purified method in different conditions: the simulated normal condition, the steady high-glucose condition and the fluctuated glucose condition, and they were intervened with S-NSAB. Thereby, the experiment was carried out in 6 groups, i.e. the normal control group (A), the S-NSAB intervened group (B), the steady high-glucose cultured group (C), the steady high-glucose cultured and S-NSAB intervened group (D), the fluctuated glucose cultured group (E), and the fluctuated glucose cultured and S-NSAB intervened group (F). Content of Glu in the extracellular fluid was detected at 24, 48 and 72 h after intervention with a full-automatic biochemical analyzer. And the data obtained were statistically analyzed with SPSS 13.0 soft ware.

RESULTSRelease of Glu at 24 h after intervention in Group E (256.33 +/- 25.73 mg/L) was obviously higher than that in Group A and Group C (134.22 +/- 9.14 mg/L and 141. 17 +/- 22.13 mg/L, P < 0.05); at 24 h and 72 h in Group B (124.50 +/- 10.30 mg/L and 30. 17 +/- 2.97 mg/L) was obviously lower than in Group A respectively (P < 0.05); in Group D at 24 h (127.50 +/- 16.94 mg/L), 48 h (26.17 +/- 3.99 mg/L) and 72 h (27.67 +/- 3.49 mg/L) were lower than in Group C; in Group F at 24 h (228.33 +/- 18.41 mg/L) and 72 h (28.00 +/- 2.41 mg/L) were lower than in Group E respectively at the corresponding time points.

CONCLUSIONSFluctuated glucose condition could obviously increase the Glu release of RGCs, to cause extracellular large amount Glu accumulation, which induces the exciting neurotoxicity to RGCs and finally to aggravate the injury on cells. S-NSAB could reduce the Glu release to some extent in the steady-high or fluctuated glucose conditions, diminish the injury of RGCs from exciting neurotoxicity of Glu, and it might be one of the intervening pathways of Chinese drugs for NSAB in preventing and treating DRP.

Animals ; Animals, Newborn ; Cells, Cultured ; Diabetic Retinopathy ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; pharmacology ; Glutamic Acid ; metabolism ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells ; cytology ; metabolism

Objective To detect the expression of survivin mRNA in cervical cancer cell lines using molecular beacon imaging technology. Methods Human cervical cancer cells (HeLa and SiHa) and human fetal lung fibroblast HFL-I were cultured in vitro. After adding 100 nmol/L survivin mRNA molecular beacon, the fluorescent signals were observed under fluorescent microscope. The expressions of survivin in cervical cancer cells and HFL-I cell were examined by immunocytochemical streptravidin-biothin peroxidase (SP) assay at the same time. Results Two kinds of survivin mRNA molecular beacon, with different color fluorescence, had strong fluorescent signal in cervical cancer cell lines, and the signal in SiHa cell line was stronger, but these signals were not found in HFL-I ; Immunocytochemical staining of positive survivin was located in the cytoplasm of cervical cancer cell lines HeLa and SiHa, whereas, no expression of survivin was detected in HFL-I cell line. Conclusion The technology of molecular beacon imaging can be used to detect the expression of survivin mRNA in viable cells successfully, and may provide a new approach to the diagnosis of early stage cervical cancer and the following-up in the clinic.

Objective To study the co-relation of radiological and histopathological features of lung cancer. Methods X-ray CT and histopathological features of 36 cases of lung cancer were analyzed retrospectively. Results High-density mass with strengthened edge was found to be paralleled with lung atery at lung gate in 6 cases. As a result, bronchial tubes became narrow with mucus in it. Squamous carcinoma tissues of 22 cases were showed growing within bronchial tubes. Local lung emphysema in early stage obstructive pneumonia and 19 cases of lung collapse in mid/late stage(86.4%) were brought out because of narrow cavity of bronchi. Megaly lymph nodes were seen by mediastina window in 12 cases. Peripheral mass with leaf-like edge were found in 8 cases,and the diameter of each was less than 2cm,while irregular deviated cavitative was seen in one case. Conlusion It is suggested that close combination of X-ray CT and histopathological can improve the accurate rate of diagnosis in lung cancer.

Adult ; Dental Implantation, Endosseous ; instrumentation ; Dental Implants ; Female ; Humans ; Orthodontic Anchorage Procedures ; instrumentation ; Orthodontic Appliance Design

OBJECTIVETo identify mouse acute pneumonia model induced by influenza virus adapted strains (FM1 strain) using RT-PCR, gene clone and sequence analysis and pathological examination of lung tissues.

METHODSAcute pneumonia was induced by intranasal drip of FM1 strain. The lungs were collected after 3, 5 and 7 days. RT-PCR was used to detect the viral load. Amplified PCR products were cloned and sequenced. Pathological and histological changes to the lungs were observed.

RESULTSThere were no abnormalities in the alveoli, alveolar sacs and alveolar septa and no inflammatory cell infiltration was found in normal mice. In the model group, we found disappearance of alveoli, alveolar sacs, alveolar ducts and alveolar septa, thickening of the alveolar septal and bronchiolar walls, and infiltration of inflammatory cells after 3, 5 and 7 days of influenza virus (IV) infection. Compared with the normal group, pathological changes at various time points were significantly increased (P<0.01). Viral nucleic acid can be detected in the lung tissue of the model group at various time points, and the pathological changes of the lung tissue were positively correlated with viral load. Sequence analysis demonstrated that there was 99.1% consistency between RT-PCR products of lung tissues in the model group and the known IV cDNA sequence (P<0.01).

CONCLUSIONSGene clone and sequence analysis may be used to identify acute mouse pneumonia model induced by FM1 strain.

Acute Disease ; Animals ; Base Sequence ; DNA, Complementary ; chemistry ; Disease Models, Animal ; Female ; Influenza A Virus, H1N1 Subtype ; Lung ; pathology ; Male ; Mice ; Molecular Sequence Data ; Pneumonia, Viral ; etiology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

To introduce professor Gao Yuchun's clinical experience and treating characteristics of facial paralysis treated with acupuncture and moxibustion. Professor Gao pays attention to yangming when he selects acupoints for clinical syndrome, and directs acupoints selection based on syndrome differentiation in different levels of jingjin, meridians and zangfu; he praises opposing needling technique and reinforcing the deficiency and reducing the excess highly; the acupuncture manipulation is gentle,shallow and slow for reducing the healthy side and reinforcing the affected side, and through losing its excess to complement its deficiency; besides, he stresses needle retaining time and distinguishes reinforcing and reducing. Facial paralysis is treated with key factors such as acupoints selecting based on yangming, acupuncture manipulation, needle retaining time, etc. And the spleen and stomach is fine and good at transportation and transformation; the meridians is harmonious; the qi and blood is smooth. The clinical efficacy is enhanced finally.
Acupuncture Points ; Acupuncture Therapy ; history ; methods ; Aged, 80 and over ; China ; Facial Paralysis ; history ; therapy ; History, 20th Century ; History, 21st Century ; Humans