With the dose of D-galactose 80mg/kg by sc administration to the female NIH mice for 6 weeks,the sub-acute aging model was established. In order to observe the effect of the polysaccharide (PSP) on anti-aging, the mice were received PSP 200mg/kg^ 100mg/kg respectively by ig administration at the same time of injection D-galactose. It was found that PSP could remarkably antagonise the effect of D-galactose on the mice. The results showed that PSP could significantly(1)decrease the content of MDA in heart,liver and brain of the mice;(2)increase the SOD activity of RBCJiver and brain in the mice;(3)increase the GSH-Px activity and GSH content of the blood and liver of the mice; (4)inhibit the MAO-B activity of liver and brain in the mice;(5)increase the hydroxyproline content of dermal tissue in aging model mice. In short,it is suggested that PSP could obviously improve the aging effect of D-galactose on the mice.

Aim ToconstructHEK293cellsstablyex-pressing corticotropin releasing factor receptor 1 ( CRFR1 ) , and evaluate its function by the cAMP as-say.Methods CulturedHEK293cellsweretransfect-ed with CRFR1-expressing vector by Lipofectamine 2000 and were selected by using G418 . CRFR1 ex-pression was detected by Western blot, RT-PCR and immunofluorescence.Results Westernblot,RT-PCR and immunofluorescence data revealed that the HEK293 cells expressed CRFR1 protein stably. The dose-responsive relationship experiment revealed that CRF induced a CRFR1-mediated cAMP production in HEK293 cells with EC50 =(5. 64 ± 0. 05) × 10 -10 mol ·L-1.Conclusion HEK293celllinesstablyex-pressing CRFR1 were constructed successfully, which would provide a cellular model to facilitate the research on the biological function of CRFR1 and CRFR1-targe-ted drug screening.

Objective To investigate the technical feasibility and clinical significance of non-vascular contrast-enhanced ultrasound(NVCEUS) in screening pyelogenie cyst out of simple renal cysts so as to avoid damage to the urinary tract from absolute ethanol while undergoing percutaneous aspiration and ethanol sclerotherapy(PAEST). Methods Following an inclusion criteria 23 patients with renal cysts were selected to receive NVCEUS scanning by means of administrating SonoVue contrast agents through puncture needle into their renal cyst lumen prior to the injection of absolute ethanol for sclerosing treatment. By the demonstration of hyperechoic contrast agents leaving from intra cyst into renal collecting system,a pyelogenic cyst was defined. The patients with this kind of cyst was not allowed for further ethanol sclerotherapy. Results NVCEUS made 3 patients with pyelogenic cyst resembling simple ones free from ethanol selerotherapy,and 4 patients suspicious of pyelogenic cyst due to weird cyst configuration remain in the list of simple cyst for further selerotherapy. Conclusions NVCEUS of renal cyst is highly capable of differentiating pyelogenie cyst from simple cyst and highly valuable in increasing the safety for the procedure of PAEST.

Objective To summarize the sonographic characteristics of parathyroid adenomas(PAs) and investigate the diagnostic values and best diagnostic thinking for an early detection. Methods Sixteen cases finally proved with PAs were retrospectively analyzed of their clinical complaints, the department for their initial consultation,indicative ultrasound findings for neck scanning, major manifestations of PAs on a series of three-mode ultrasound imagings,association of adenoma sizes to serum parathyroid hormone(PTH) levels. Results On multimode ultrasound images, the PAs were multidisciplinary morphologic, homogenously hypoeehoic,absent of calculus and necrosis,highly vascularized with color signals,and similar to thyroid in contrast agents perfusion. Serum PTH levels were elevated in accordance with increase of adenomas' size. Among the 16 cases 12 were defined as with parathyroid incidentalomas, to which liver or/ and kidney stones were contributed as indicatives. Conclusions High frequency ultrasound with multiple imaging modes are most suitable for scanning and detection of neck PAs. For those with stones in liver and urinary tracts, unknown bone fracture etc, ultrasound scanning of the neck parathyroids used to reveal latent PAs.

AIM To investigate the effects of tranfection of IL 2R antisense RNA expression plasmids on mouse spleen cells' proliferation in vitro and its possible mechanism. METHODS Spleen cells were transfected with IL 2R antisense RNA eukaryotic expression plasmids using adhesion assisted lipofection method, and then the spleen cells were stimulated by mitogen. Cells' proliferation was tested by tetrazolium salt (MTT) method. IL 2R mRNA and protein expression level were measured by slot blot hybridization assay and flow cytometry method respectively. RESULTS The proliferation of spleen cells was inhibited obviously after transfecting with recombinant plasmids. The inhibitory rate of pcAnti mIL 2R?? and pciAnti mIL 2R?? transfected group was higher than that of pcAnti mIL 2R? and pcAnti mIL 2R? transfected group; the inhibitory rate of pcAnti mIL 2R? tranfected group was higher than that of pcAnti mIL 2R? tranfected group. No inhibitory effect on the growth of NIH3T3 cells was observed when they were transfected with recombinant plasmids. IL 2R mRNA and protein expression level were decreased in spleen cells after transfection of recombinant plasmids. CONCLUSION IL 2R antisense RNA can efficiently inhibit the proliferation of mouse spleen cells in vitro. IL 2R?? chimeric antisense RNA showed higher inhibitory rate than ? or ? antisense RNA. IL 2R? antisense RNA was more effective than ? antisense RNA. It can be concluded preliminarily that the inhibitory effect of IL 2R antisense RNA was exclusively on the growth of cells functionally expressing IL 2R. The inhibitory effect on the spleen cells proliferation was likely due to the blocking of IL 2R expression by antisense RNA.

Antitumor effects of erythromycin and the related mechanism were investigated in the present study. Neuroblastoma cells (SH-SY5Y) were exposed to erythromycin at different concentrations for different durations. Cell proliferation was measured by cell counting, and cell viability was examined by MTT assay. Cell cycle phase distribution and the cytosolic calcium level were detected by flow cytometry. Mitochondrial membrane potential was measured by the JC-1 probe staining and fluorescent microscopy. The expression of an oncogene (c-Myc) and a tumor suppressor [p21 (WAF1/Cip1)] proteins was analyzed by using Western blotting. Erythromycin could inhibit the proliferation of SH-SY5Y cells in a concentration- and time-dependent manner. The cell cycle was arrested at S phase. Mitochondrial membrane potential collapsed and the cytosolic calcium was overloaded in SH-SY5Y cells when treated with erythromycin. The expression of c-Myc protein was down-regulated, while that of p21 (WAF1/Cip1) protein was up-regulated. It was concluded that erythromycin could restrain the proliferation of SH-SY5Y cells. The antitumor mechanism of erythromycin might involve regulating the expression of c-Myc and p21 (WAF1/Cip1) proteins.

Aiming at the characteristics of biochemistry and the special advantage of flash and basing on teaching practice,we set up a set of biochemistry flash animation database,providing a series of immediate flashes and being applied to teaching practice of biochemistry,which shows the great potential of flash in improving biochemistry teaching quality.

Objective To explore the feasibility of adding a flexible linker between two-pore-domain potassium channel TREK-1 (TWIK related K + channel 1)monomers to construct a tandem-linked dimer.Methods PCR was used to add a flexible linker between the two TREK-1 monomers.The cRNA obtained from in vitro transcription using the above vector was injected into Xenopus oocytes.After 24 -48 h,currents were recorded from these oocytes using a two-electrode voltage clamp.The effects of extracellular Ba2 + and pH on TdTREK-1 were observed and compared with those of native dimeric TREK-1.Results The tandem-linked dimeric TdTREK-1 was highly expressed in Xenopus oocytes.The currents through these channels were inhibited by extracellular Ba2 +and acidification.Furthermore,the responsiveness of the concatenated dimers to these extracellular stimuli was similar to that of native dimers.Conclusion Adding a flexible linker between the two monomers to construct the tandem-linked dimer does not affect the expression and gating properties of TREK-1, suggesting that the method be feasible.Such a method will allow the manipulation of a single subunit,which will help basis study the structure and function of TREK-1.

AIM: Open reading frame(ORF) of death associa ted protein kinase1(DAPK1) gene was cloned for studying on tumor forming and met astasis.METHODS: Based on nucleotide sequence of DAPK1 gene f rom GenBank, a pair of primers was designed. DAPK1 gene ORF was transfected into Raji cells in expression vector pcDNA3.1(+) with lipofectamine reagent. Morphol ogic assessment of apoptosis was performed with fluorescence microscope cytotoxi city and cell viability was assayed by MTT. RESULTS: DAPK1 gene ORF was amprified from K562 cells by RT-PCR. It was cloned into plasmid pMD18-T and sequenced. There were seven mutation in 4 300 bp nucleotide sequence rel ativel y to DAPK1 nucleotide sequence from GenBank, but six was synonymous mutation and one was single nucleotide polymorphism. 4 300 bp nucleotide of DAPK1 gene O RF was transfected into Raji cells. DAPK1 gene expression was detected in 48 h a fter it was transfected into Raji cells. Then Raji cells showed apoptosis.CONCLUS ION: Large fragment gene was cloned by RT-PCR and transfected into Raji cells successfully. Over-expression of DAPK1 gene induced Raji cells apoptosis.

AIM: To inhibit the expression of tyrosinase gene in murine B16 melanoma cells by antisense nucleotide. METHODS: The antisense recombinant pcDNA3.1(-)-tyr was constructed and was used to infect murine B16 melanoma cells for expression of tyr antisense nucleotide. The effect of antisense nucleotide of tyr on the expression of tyr gene was detected by determination of the activity of tyrosinase and of the production of melanin, Dopa staining and electronic microscope. RESULTS: The tyr antisense recombinant was successfully constructed and injected into murine B16 melanoma cell. The activity of tyrosinase in B16 cells infected with pcDNA3.1 (-)-tyr decreased to 0.0498?0.0036, compared to the tyrosinase activity of 0.0916?0.0132 in the control cells without treatment (P