Chlamydial infection in human urogenital tract induces inflammation and causes tissue damage and scarring. It is thought that cytokine production by the Chlamydia-infected cells plays a key role in chlamydial disease processes. Although many cytokines have been detected during chlamydial infection, little is known about the molecular mechanisms on how Chlamydia triggers and sustains the inflammatory cytokine cascades. In the current study, chlamydial infection of the human cervical epithelial cell line HeLa cells can induce the production of IL-8, IL-1α, IL-1β and IL-6. Using inhibitors for probing intracellular kinase signaling pathways required for the Chlamydia-induced cytokines, it was found that the Chlamydia-activated MAPK / P38 pathway is required for the chlamydial induction of IL-1α and IL-6 while both the Chlamydia-activated MAPK/ERK and MAPK/P38 pathways contribute to the production of IL-8.

Objective To improve diagnosis accuracy of pineocytoma (PC) by joint analysis of CT,MRI imaging features and differential diagnosis with other lesions in pineal region.Methods Totally 6 pineocytoma patients confirmed surgically and pathologically had their clinical history,CT and MRI data collected and analyzed on lesion morphology,cystic solid changes,existence of necrosis,complications of hemorrhage and or calcification,MRI and enhanced scan of solid component,complications with hydrocephalus and etc.Results Plain scan found 1 case of solid nodule and 5 cases of cystic-solid nodules,2 cases with clearly-bordered lesions and 4 one not as well as 4 cases with significant hydrocephalus and 2 ones with light hydrocephalus.Enhanced scan showed 5 cases of moderate to marked enhancement and one case with no obvious enhancement.CT examination proved there were 1 case of calcification and 1 case of hemorrhage.Conclusion Pineocytoma has the characteristics of benign tumor,and has to be differentiated with other tumors frequently occurring in this region in case of obvious clinical signs due to crushing brain parenchyma or blocking aqueduct cerebri by oversized lesions.

Objective To establish an HPLC method for simultaneous separation and determination of chlorogenic acid, quercetin and kaempferol inPyrrosia lingua (Thumb.) Farwell. Methods The separation was performed on Phenomenex C18 column (250 mm×4.6 mm, 5μm) with the mobile phase of methanol-acetic acid (pH=3.0) solution and gradient elution. Flow rate was 1.0 mL/min;the UV detection wavelength was 254 nm;column temperature was 35℃.Results The calibration curves for chlorogenic acid, quercetin, and kaempferol were in good linearity in the range of 0.000 24-3.00μg (r=0.999 9), 0.000 16-2.00μg (r=0.999 9), and 0.000 16-2.00μg (r=0.999 9), respectively. The limits of detection (S/N=3) were 3.29, 0.43 and 0.33 ng/mL, respectively. The average recovery rates were 97.73%, 98.07% and 96.92%, respectively. ConclusionThe method is simple, precise and sensitive. It provides scientific proof for separation and determination of chlorogenic acid, quercetin and kaempferol inPyrrosia lingua (Thumb.) Farwell.

This study was aimed to observe the influence of water-solubility nipponica saponin on activation of TNF-α+IL-17-induced rat fibroblast-like synovial cell line RSC-364 cellular model nuclear transcription factor NF-κB pathway as well as TNF-α, IL-1, ICAM-1, MMP-2, MMP-3 secretion. IL-17+ TNF-α were used for stimulating RSC-364 to establish rheumatoid arthritis (RA) cellular model. Water-solubility nipponica saponin in different con-centrations was used for intervention. The influence of water-solubility nipponica saponin in different concentrations on cell viability was detected by semi-quantitative RT-PCR method. Changes in the level of TNF-α, IL-1, ICAM-1, MMP-2, and MMP-3 of culture supernatant were detected by ELISA. The results showed that the activation of NF-κB p65 in RSC-364 stimulated by TNF-α+ IL-17 can be inhibited by water-solubility nipponica saponin ac-cording to its concentration. It improved IκB-α expression, and inhibited TNF-α, IL-1, ICAM-1, MMP-2 and MMP-3 secretion. It was concluded that water-solubility nipponica saponin can inhibit the activation of NF-κB pathway, hinder the secretion and activation of multiple downstream genes, which may be its effect in inhibiting syn-ovial inflammation in RA.

Objective To overcome the fact that SRV-NM virus can only multiple and amplify through partially pu-rified jaagsiekte retrovirus inoculated intratracheally in sheep but it cannot be augmented using in vitro cell culture, we con-structed JSRV-NM pseudovirions based on high efficiency packing system of lentivirus. Methods Lentivirus of three high efficiency packing plasmids system pMD.G, pCMV-HIV 8.2 and pHIV-eGFP was developed, and JSRV-NM-env coated plasmid pCMVJSRV-NM was used to substitute VSV-G virus coated plasmid pMD.G then co-transfected into 293T cells to replicate, package and produce restructured JSRV-NM pseudovirions. Gene expression of pseudovirion was determined through WPRE using real time PCR; Virus infectivity was detected through inoculating JSRV-NM pseudovirions into 24 pore plates. Results We construct JSRV-NM pseudovirions successfully based on the lentivirus system. JSRV-NM pseudo-virions can also be concentrated to higher titer (108 TU/mL detected by real time PCR by ultracentrifugation without signifi-cant loss of activity. JSRV-NM and VSV-G pseudovirions infected on Hela cells (both MOI= 3) respectively and no obvi-ous difference were shown on their infection efficiency detected by real time PCR. Conclusion Based on lentivirus system, JSRV-NM pseudovirions can be multipled and amplified in 293T cell culture in vitro. JSRV-NM pseudovirions is stable without loss its infection activity and the requirements of biological laboratory safety II was also met. JSRV-NM pseudoviri-ons will provide a useful tool for further study of JSRV-NM-env infection across species or its induction of lung adenocarci-noma.

Aim To screen antibodies of novel purinoceptor as a marker for further study of the purinoceptor. Method BALB/c mice were immunized for 4 times with rat aortic endothelial cell. Then the phage display system was used to construct a single-chain Fv fragment (ScFv) cDNA library from the total RNA of immunized mice. The characteristics of novel purinoceptor not existing on vascular smooth muscle cell but on aortic endothelium were used to enrich the aortic endothelium specific antibodies. Induced with IPTG, these antibodies were secreted into the periplasm of E. coli. The functional experiment of novel purinoceptor named organ bath experiment was used to screen out the positive ScFv from the soluble expressed antibodies. Immunohistochemistry experiment was used for positive ScFv identification. Results The total mouse anti-rat endothelium lgG is 1 ∶16 000. 8?106 mouse anti-rat endothelium ScFv cDNA library was successfully constructed. After 4 times of rat endothelium and rat smooth muscle cells screening, 2 500 ScFv cDNA binding membrane of aortic endothelium was enriched. After 4 times of functional screening, a phage-ScFv named B inhibiting the adenosine induced NO dependent construction by 83.4%?21.6% was selected from the expressed antibodies. Immunohistochemistry experiment showed that ScFv-B combined with aortic endothelium specifically and functional experiment showed that ScFv-B did not have any effect on adenosine induced ileum contraction, indicating that ScFv-B specifically binding to the novel purinoceptor. Conclusions ScFv-B binding specifically to the novel purinoceptor was selected by phage display technique and functional screening experiment which provide a good marker for further study of the novel purinoceptor.

Comparison of the institutional setup, policies and adverse event report mechanism for medical risk control in the countries of UK, USA, Canada, and Australia by means of browsing information on their official websites. It is found that these countries maintain a national patient safety authority, coupled with a tiered management at national, local, medical institutions and NGOs level; the USA pattern features laws and regulations, that of UK and Australia features guidelines as policy guarantee for medical safety; these countries regulate adverse event reporting by either government leadership or cooperation with trade associations. Inspirations from this study suggest China to enhance institutional construction, complete regulations, and advocate the culture for medical safety, and to build the national-level reporting and study system for medical safety events, and improve medical risk management.

A novel double-chamber series piezoelectric pump has been presented and tested. The pump is a multi-layer circular planar structure, consisting of PMMA (polymethyl methacrylate) pump body, two PZT actuator membranes and three cantilever valves. The PZT actuators are driven at a phase difference of 180 degrees, which is equal to two one-chamber pumps running in series. The output performance depends on the geometrical parameters of the actuator membrane. The prototype pump, fabricated with the PZT membrane 0.18 mm in thickness and 50 mm in diameter of 50 mm, can deliver drug in either direct way (pumping liquid drug) or indirect way (pumping air to extrude liquid drug from a sealed container). The frequency-response characteristic of the two handling methods is of difference. The pump obtains optimum performance at low frequency for liquid as medium, and at high frequency for air as medium. For both the direct delivery and indirect delivery, the maximum flowrate achieved reached up to 220 ml/min and 35 ml/min, respectively; and the maximum backpressure obtained amounted to about 14 KPa and 21 KPa, respectively, at the applied voltage of 80 V with frequency of 20 Hz.
Drug Delivery Systems ; instrumentation ; Electricity ; Equipment Design ; Evaluation Studies as Topic ; Insulin Infusion Systems ; Models, Theoretical

OBJECTIVE: To determine the signaling pathway required for Chlamydial induction of IL-8 expression in epithelial cells. METHODS: The production and localization of IL-8 in Chlamydia-infected Hela 229 cells were monitored using Western blot, immunoflourescence, and ELISA. Activation of MAPK and NF-kappaB signaling pathways were detected by Western blot and immunoflourescence. The effect of different signaling pathways on Chlamydia-induced Il-8 was measured by experiments of chemical inhibitors. RESULTS: IL-8 was induced by Chlamydia and was time-dependant. Chlamydial infection activated MAPK/ERK and MAPK/p38 pathways but not NF-kappaB pathway. Chlamydial induction of IL-8 was blocked by small molecule inhibitors targeting the ERK and p38 pathways. CONCLUSION Chlamydia-induced IL-8 in cervical epithelial cells, the natural target cell type of Chlamydia trachomatis infection, is dependent on MAPK pathway but not NF-kappaB pathway, which provides important information for further understanding the molecular mechanism of Chlamydia-induced inflammatory pathologies.
Chlamydia Infections ; metabolism ; Chlamydia trachomatis ; physiology ; Epithelial Cells ; metabolism ; microbiology ; HeLa Cells ; Humans ; Interleukin-8 ; biosynthesis ; NF-kappa B ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism

With a microsystem or micropump, the release rate of drug delivery is able to be controlled easily to maintain the therapeutic efficacy. A piezoelectric membrane-valve micropump for implantable and carryhome drug delivery system is developed and tested. The influence elements of dynamic performance of the PZT actuator and valve were analyzed, and the calculation method of resonant frequency of the membrane valve was provided. Study results showed that the output performance of the micropump depended on the coupling effect of the actuator and valve. For a given actuator, the output value and the optimal frequency of a micropump could be enhanced only by valve design. Two micropumps with different valve dimensions were fabricated for comparing examination. The smaller -valve micropump obtained higher output values (the maximum flow rate and backpressure being 3.5 ml/min and 27 KPa, respectively) and two optimal frequencies (800 Hz and 3 000 Hz). The larger -valve micropump achieved lower output values (the maximum flow rate and backpressure being 3.0 ml/min and 9 KPa, respectively) and one optimal frequency (about 200 Hz). The test results suggest that the output values and optimal frequency of micropump can be improved by changing the valve dimension, and the viewpoint that checkvalve micropump works only with low acting frequency is wrong.
Drug Delivery Systems ; instrumentation ; Equipment Design ; Humans ; Infusion Pumps, Implantable ; Micro-Electrical-Mechanical Systems ; instrumentation