Objective To explore the dynamic changes of blood brain barrier(BBB)permeability after experimental subarachnoid hemorrhage(SAH)in rats.Methods Eighty female SD rats were divided into normal saline control group(n=10)and SAH group(n=70).The SAH model was induced by injecting 300 ?l of autologous arterial blood into the subarachnoid space near the circle of Willis via a cannula in an artificial hole between the olfactory bulb and frontal lobe.BBB permeability in cerebral cortex of SAH and normal saline rats was assessed at 6,12,24,36,48,60,and 72 h after SAH establishment by fluorescence spectrophotometer and fluorescence microscopy for Evans Blue(EB)extravasation.The ultrastructural changes in BBB were observed with transmission electron microscope.Results Compared with the control group,the changes of Evans Blue content and extravasation in cerebral cortex of SAH group peaked at 36 h(P0.05).The severe ultrastructural abnormality was found at 36 h after SAH.Conclusion The changes of BBB permeability develop at the acute stage of SAH,resulting from multiple factors together.The BBB after SAH possesses a self-repairable property.

The major challenge in the development of recombinant biologics lies in generating and isolating rare high-producing stable single clone in a short period of time. The selection marker is an essential component of the plasmid vector, it plays an important part in the generation and screening of producing cell lines. Engineering the selection marker to enhance the stringency of selection for high producing cells is one of the most effective approaches to improve the cell line development process. Here, using Chinese hamaster overy (CHO) cells as an example, we introduce the application of selection marker for generation of recombinant biologics producing mammalian cell lines, methods of engineering the selection markers to enhance the selection stringency, and propose considerations on cell substrate stability and selection marker safety, in order to provide references for high-efficiency development of recombinant biologics.

Objective To explore the effects of transcranial magnetic stimulation (TMS) on motor function in patients with Parkinson's disease (PD) using meta-analysis. Methods Eight comparative studies of the effects of TMS were meta-analyzed. Results The combined studies confirmed a significant difference before and after TMS treatment. Between the experimental and control groups the effect was also highly significant. Conclusion TMS may play an active role in the rehabilitation of motor function for patients with Parkinson's disease.

Animals ; Cell Proliferation ; drug effects ; Female ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tamoxifen ; analogs & derivatives ; pharmacology

Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

Objective To assess the biophysical characteristics of normal skin and inflammatory skin lesions in mild and moderate acne. Methods Seventy-five mild and moderate acne vulgaris patients were included in the study. One inflammatory lesion measuring 2.0 to 5.0 mm in diameter which occurred within 48 hours prior to the measurement was selected as the target lesion. Trans-epidermis water loss (TEWL), capacitance and a* value were measured in target lesions and lesion-adjacent normal skin. Sebum content immediately,1 and 4 (saturated) hours after face washing was also determined on target lesions and central forehead between the eyebrows. Sebum secretion rate (SSR) was calculated. Results The TEWL and SSR significantly increased in the lesion-adjacent normal skin with the increment of inflammatory lesion number (both P < 0.05), and increased in target lesions with the elevation of a* value (both P < 0.05). The target lesions exhibited a significantly higher TEWL but a lower saturated sebum content and SSR than the adjacent normal skin did (all P <0.05). Conclusions The severity of inflammatory lesions is correlated with the impaired barrier function and increased SSR in facial skin. Compared with the adjacent normal skin, inflammatory skin lesions have a reduced skin barrier function and SSR.

A poly( GMA-EGDMA) coated SPME fiber was prepared using an in-situ polymerization by direct bonding to the surface of a polydopamine-modified stainless steel wire. Then the fiber was modified by sulfuric acid. A novel solid phase microextraction coating coupled to high performance liquid chromatography ( HPLC) method based on the as-prepared fiber was developed for the determination of four pharmaceuticals and personal care products ( PPCPs) in water samples. The influences of extraction parameters, including pH, extraction time, extraction temperature and salt addition were investigated. 3 mL water sample was extracted by the as-prepared fiber for 60 min at 30 ℃, and then desorbed with mobile phase for 30 min, respectively. Desorption solution was analyzed by HPLC-DAD ( diode array detection ) . The results indicated that the extraction yield of the fiber was good for four PPCPs. The linear correlation coefficients were>0. 997 with the linear range of 2-200 μg/L. The limits of detection (S/N=3) were 0. 5-5 μg/L with RSD (n=5) of 4. 1%-11. 9%. The recoveries of four PPCPs at spiked level of 20, 50, 100 μg/L were within the range of 70. 6%-105. 5%. The results showed that this method was easy, green, accurate and precise, and could be used to assay the four PPCPs in real water samples.

Objective To investigate the roles of activin A(ACT-A) and follistatin(FS) in renal interstitial fibrosis.Methods Renal interstitial fibroblasts were isolated from SD rat kidney and primarily cultured.The cells were divided into A group,F group and A+F group,and then each group was further divided into 5 subgroups according to their culture media being added with ACT-A(0,0.3,3,30,or 100 ng/ml),FS(0,0.3,3,30,or 100 ng/ml),and ACT-A(30 ng/ml) plus FS(0,0.3,3,30,or 100 ng/ml) respectively.Levels of type Ⅳ collagen(Ⅳ-C) and hydroxyproline(HYP) in the supernatants of cultured fibroblasts were measured by ELISA assay.Results Concentrations of Ⅳ-C and HYP in cultured rat renal interstitial fibroblasts were in a dose-dependent manner with ACT-A treatment.FS had no effect on these concentrations,but it inhibited the effects of ACT-A on Ⅳ-C and HYP in the supernatants of cultured fibroblasts in a dose-dependent manner.Conclusion ACT-A might be involved in occurrence and development of renal fibrosis by affecting renal interstitial fibroblasts.Exogenous FS could suppress the effects of ACT-A on the cultured renal fibroblasts.

Objective To study the effects of cholecystokinin (CCK) on canine Oddi sphincter(SO)function after pancreas transplantation with bladder drainage. Methods Normal canine and transplant canine SO manometry before and after CCK administration was carried out. Results SO basal pressure in control group was(18 5?2 8) mm Hg, frequency was(9 7?1 5)per min, amplitude was (47?6) mm Hg, motility index was(236?56). After CCK administration, basal pressure, frequency, amplitude and motility index decreased significantly to(10 2?2 2) mm Hg ,(5 0?1 2)per minute,(19?5) mm Hg and(50?17), all P

OBJECTIVE: To investigate the expression of monocyte chemotactic protein-1 (MCP-1) and its receptor CC chemokine receptor-2 (CCR-2) in coronary atherosclerosis plaques between sidden coronary death (SCD) and non-SCD. Methods The expression levels of MCP-1 and CCR-2 in SCD group, coronary atherosclerosis group (non-SCD), control group (normal coronary artery) were detected by immunohistochemistry. RESULTS: Positive rates of MCP-1 among the three groups were 78%, 47%, and 0%, respectively, with significant expressing differences between each two groups (P<0.05). Positive rates of CCR-2 among three groups were 72%, 47%, and 0%, respectively, with significant expressing differences between the SCD group and coronary atherosclerosis group as well as between the SCD group and control group (P<0.05), but with no significant expressing difference between coronary atherosclerosis group and control group (P>0.05). CONCLUSION Overexpression of MCP-1 and CCR-2 in coronary atherosclerotic plaques is closely correlated with SCD.
Chemokine CCL2/metabolism* ; Coronary Artery Disease/pathology* ; Death, Sudden, Cardiac/pathology* ; Humans ; Immunohistochemistry ; Receptors, CCR2/metabolism*