Objective To investigate the possible mechanism of glomerular injury in diabetes mellitus by determining whether epithelial-mesenchymal transition (EMT) is caused by high glucose in mice podocytes. Methods Using mice glomerular podocyte cell line as an in vitro system, podocytes were incubated with glucose(12.5 mmol/L, 25 mmol/L, 50 mmol/L) and mannitol (50 mmol/L) for 36 hours. Then the cells were collected and expression of alpha-smooth muscle actin(α-SMA), fibronectin (FN), CD2 associated protein (CD2AP) and Wilms' tumor 1 gene (WT-1) was detected by Western blot and indirect immunofluorescence staining. Results Under low glucose (5.6 mmol/L) and mannitol (50 mmol/L) condition, there were high expression of CD2AP and WT-1, and low expression of α-SMA and FN in mice podocytes. After 36 hours treatment with high glucose (12.5 mmol/L), the expression of α-SMA and FN in podocytes was significantly increased, and the expression of α-SMA and FN was further up-regulated with the increase of glucose dosage (25, 50 mmol/L). The indirect immunofluorescence staining revealed the similar result, and the percentage of positive α-SMA cells was also increased compared with low glucose and mannital group (P<0.05). Meanwhile, Western blot showed that high glucose could down-regulate the expressions of CD2AP and WT-1 in a dose-dependent manner. Conclusion EMT may be a potential pathway leading to podocyte dysfunction and glomerular injury under high glucose conditions.

Objective To explore an economic and effective management strategy for lumbar disc herniation (LDH) in community health center. Methods 62 LDH patients treated in Xujiahui community health center (observation group), 48 LDH patients treated in Huajing community health center (control group A) and 55 LDH patients treated in Shanghai 6th People's Hospital (control group B) were included.The observation group was managed by associate professor from Shanghai 6th People's Hospital and accepted different treatment strategy in mild, medium and heavy grades. The control groups A and B were managed routinely. 3 month later, costs of illness were computed and curative effects were accessed in 3 groups. Results There was no difference in curative effects between observation group and control group B (P>0.05), but better than control group A (P<0.05). The observation group cost the least among the groups (P<0.05). Conclusion Graded treatment strategy united with tertiary hospital is an economic and effective mode for LDH management in community health center.

The mosquito vector of filariasis malayi and its vectorial capacity was investigated In 5 endemic villages in Leshan Prefecture, Sichuan Province. The results showed that the man-biting rate, numan blood index and vectorial capacity of An. lesteri anthropophagus were 0.7, 5.1 and 10.63 times higher than those of An. sinensis. Besides, the natural infection by microfilaria in An, lesteri anthropophagus was also higher than that in An. sinensis by 5 times.From the above result, the authors concluded that An. lesteri anthropophagus was the main vector for transmitting filariasis malayi in the area under study.

Perimenopausal symptom especially hot flash has a serious effect on women ’ s mental health and life quality.Selective serotonin reuptake inhibitors ( SSRI ) are medicines which inhibit reuptake of sero-tonin thus increasing the concentration of serotonin on synaptic cleft.These medicines are used for treating dipression.Recent studies found that they could also alleviate perimenopausal hot flash.In this paper we will review the effect of SSRIs on treating hot flash.

Objective:To investigate the effects of musk-1, a glucoprotein component isolated from the water extract of musk, on some functions of rat polymorphonuclear leukocytes activated by IL-8 in vitro.Method：An in vitro incubation system was used. Superoxide anion production was determined by cytochrome C reduction. β-glucuronidase and lysozyme release was quantitated by enzyme reactions in which phenolphthaleinglucuronic acid and Micrococcus Lysodeikticus were as the substrates, respectively.Results:In comparison with control, musk-1 at concentration 1～100 μg*ml-1 can increase superoxide anion production by 91.7%～291%, and decrease β-glucuronidase and lysozyme release by 2.2%～58.1% and 3.9%～39.8%, respectively.Conclusion:Inhibition of lysosomel enzyme release might be considered as one of mechanisms of antiinflammatory action of musk.

Aim To investigate the effects of salidroside on NeuN and Egrs in the ischemic side of middle cerebral artery occlusion (MCAO) rats by inhibiting complement C3.Methods The rats were subjected to MCAO with suture-occluded method,and the neurologic injury was evaluated.The rats underwent l h ischemia/reperfusion with 1 d and 2d salidroside treatment,and the expressions of NeuN,Egr4,C3 and C1 were tested.Male Sprague Dawley rats were separately injected ventricle C3aR inhibitor and artificial cerebrospinal fluid with the help of ventricle stereotaxic apparatus.Thirty min later,the models of MCAO were finished with 1h reperfusion,drug administration and intracerebroventricular injection for 2d.The expressions of NeuN,Egr4,C3 were detected.Results Compared with models of MCAO,the expression of C3 in MCAO rats treated with salidroside 1 d and 2d decreased significantly,and the expression of NeuN increased markedly.Salidroside had no apparent effect on Egr4 and C1 of administration of 1d,but it could significantly enhance the expression of Egr4 after 2d,and reduce the expression of C1 significantly after 2d.The rats administrated with C3aR inhibitor into cerebral ventricle continueously showed the same result in accordance with the treatment of salidroside.And the treatment of salidroside and C3aR inhibitor did not show remarkable additive effects.Conclusion The neuroprotective effect of salidroside on acute cerebral ischemia/reperfusion injury may be related to the inhibition of the activation of the classical pathway of complement,the regulation of Egrs and the reduction of apoptosis.

OBJECTIVETo suppress the expression of CCR5 and CXCR4, the co-receptors for human immunodeficiency virus type 1 ( HIV-1), and thus inhibit HIV-1 from entering cells.

METHODSDNA fragments encoding either CCR5 or CXCR4 were amplified from healthy human peripheral blood mononuclear cells (PBMCs) by reverse transcript polymerase chain reaction (RT-PCR) and sequencing was performed. Correct fragments were inserted into Shuttle plasmid inversely, which was recombined with backbone plasmid containing homologous adenoviral genome in E. coli BJ5183. The recombinant plasmids were transfected into 293 cells in which they were packaged and amplified. Recombinant adenoviruses containing antisense RNA of CCR5 or CXCR4 were obtained and identified by RT-PCR, and the titres of them were determined by cytopathic effect (CPE) method. The U937 and MT4 cells were infected by recombinant adenoviruses containing antisense RNA of CCR5 (multiplicity of infection, MOI = 100) and CXCR4 (MOI = 200), respectively. The expression of co-receptors on infected cell was measured by fluorescence activated cell sorter at 24, 48, 72 hours and 10 days after infection. In addition, the chemotactic activity and proliferation of infected cells were detected with Boyden chamber and 3H incorporation respectively.

RESULTSWe constructed the recombinant plasmids and obtained the recombinant adenoviruses which contained antisense RNA of CCR5 or CXCR4 and were designated as pAd-antiR5 and pAd-antiX4 respectively. The titers of recombinant adenoviruses pAd-antiR5 and pAd-antiX4 were 5 x 10" PFU/ml and 7 x 10(10) PFU/ml, respectively. The expression rate of CCR5 on U937 cells decreased from 82. 10% (blank control) to 1.12% (Ad-antiR5 infected) , and that of CXCR4 on MT4 cells decreased from 42% (blank control) to 1.03% (Ad-antiX4 infected) 24 hours later. The expression rates of CCR5 on Ad-antiR5 infected U937 cells were 1.02% , 1.26% , 1.23% at 48 hours, 72 hours, and 10 days later, respectively. The expression rates of CXCR4 on Ad-antiX4 infected MT4 cells were 1.13%, 1.17%, 1.22% at 48 hours, 72 hours, and 10 days later, respectively. Moreover, the recombinant adenovirus had no effects on chemotactic activity and proliferation of the cells.

CONCLUSIONThe recombinant adenovirus containing antisense CCR5 or CXCR4 can remarkably decrease the expression of co-receptors for HIV-1 on U937 or MT4 cells without affecting their chemotactic activities and proliferative abilities.

Adenoviridae ; genetics ; Cell Line, Transformed ; Cell Proliferation ; Chemotaxis ; Down-Regulation ; Genetic Vectors ; Humans ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics ; Receptors, CCR5 ; biosynthesis ; genetics ; Receptors, CXCR4 ; biosynthesis ; genetics ; Transfection ; U937 Cells

OBJECTIVETo purify mutacin produced from isolated Streptococcus mutans (S. mutans) strains in order to, contribute to molecular biological research of mutacin.

METHODSThe antibacterial activity of 80 isolated strains was tested by the stab culture technique against Streptococcus oralis ATCC 10557. The mutacin produced by strain 1G, was initially purified by solid-phase extraction (SPE) after crude extraction by chloroform. And then the active substances were purified by twice reversed-phase high performance liquid chromatography (RP-HPLC). The purified target peptide (mutacin) was collected and freeze-dried for further study.

RESULTSThe greatest active strain of these S. mutans isolates, the strain 1G was obtained. Roughly 15 microg crude mutacin was extracted from 200 mL liquid medium of this strain 1G. The purified mutacin through SPE and twice RP-HPLC was obtained.

CONCLUSIONIt was much complex to separate and purify mutacin due to its small molecular mass, and extracting and purifying of mutacin may make an important contribution to the further research of mutacin.

Anti-Bacterial Agents ; Bacteriocins ; Streptococcus mutans

Two strategies, direct ligation after enzyme digestion and over-lap PCR technology, were adopted to construct a fusion gene which was composed of the antimelanoma single chain antibody gene and the staphylococcal enterotoxin A gene without N-terminal signal sequence. The fusion gene was subcloned into pET28-a vector and transformed into E. coli BL21(DE3). Ni-NTA system was selected to separate and purify the expresstd products. The inhibition ratio of the fusion protein was tested by MTT method. It is shown that the 6His-ScFv-SEA fusion protein can be expressed stably in E. coli BL21 (DE3). The quantity of the fusion protein was shown up to 30% of the total protein of the bacteria and mainly in inclusion body. By activation the effective cells, the fution protein can inhibit the melanoma cell whith expressed corresponding antigen.
Cell Line, Tumor ; Cell Survival ; drug effects ; Cells, Cultured ; Electrophoresis, Polyacrylamide Gel ; Enterotoxins ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Humans ; Inclusion Bodies ; genetics ; metabolism ; Melanoma ; drug therapy ; immunology ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; therapeutic use ; Single-Chain Antibodies ; genetics ; metabolism

AIMTo explore the physiopathological mechanisms of airway injury and the effect on the airway responsiveness of rat by inhaled sulfur dioxide(SO2).

METHODSSixteen SD male rats were divided randomly into 2 groups (n = 8): the control group and SO2 group. The control group was exposed o pure air. SO2 group was exposed to SO2 of the content 1.0 mg/(m(3) x h) 6h daily for consecutive 3 d. At 4th day, we determined the airway responsiveness, collected the bronchoalveolar lavage fluid (BALF), plasma and lung tissue. Then we counted the total cellular score in BALF, measured the plasma SP content and made the immunohistochemistry staining on the lung tissue (HE and SP methods).

RESULTSCompared with the control group, the total cellular score in BALF and plasma SP content in SO2 group's increased significantly ( P < 0.01). HE staining showed there were a great deal of inflammatory cells infiltration under the tunica mucosa bronchiorum; and SP immunohistochemistry staining indicated there were significant changes in numbers of SP-IR positive fibers of SO2group.

CONCLUSIONExposure to low concentration of SO2 would injure healthy rat's airway, and induce airway hyperresponsiveness, neurogenic inflammation is one of its critical pathophysiological mechanisms.

Air Pollutants ; adverse effects ; Animals ; Asthma ; chemically induced ; Bronchi ; drug effects ; innervation ; physiopathology ; Bronchial Hyperreactivity ; chemically induced ; physiopathology ; Bronchitis ; chemically induced ; Bronchoalveolar Lavage Fluid ; cytology ; Male ; Nerve Fibers ; drug effects ; physiology ; Neurogenic Inflammation ; chemically induced ; physiopathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Substance P ; blood ; Sulfur Dioxide ; adverse effects