One major problem to successful treatment of cancer is the development of resistance by tumor cells to multiple chemotherapeutic drugs, a phenomenon named multidrug resistance (MDR). Searching for the novel chemotherapeutical agents is one of the important strategies for overcoming MDR. By using a cytotoxicity assay, flow cytometry analysis, Western-blotting and RT-PCR, a drug (Taxol, TAX) resistant human nasopharyngeal carcinoma KB cell line (KB/TAX) was established by addition of the drug to the cell cultures gradually, then a novel N-sugar substituted thalidomide analogue (STA-35) was investigated for its reversal effect on MDR of KB/TAX cells and possible mechanism. The results showed that KB/TAX cells were resistant to several chemotherapeutical agents, and the relative resistance to TAX was 73.1. Compared with parental KB cells, the function and protein expression of P-glycoprotein (P-gp), as well as mdr1 gene in the KB/TAX cells were remarkable reduced. Moreover, both KB and KB/TAX cells were sensitive to STA-35, the relative resistance to TAX on KB/TAX cells was decreased by the addition of STA-35. Furthermore, STA-35 (5 ～20 ?mol/L)was capable to reduced the activity of P-gp by increasing the accumulation of rhodamine 123, decreasing P-gp expression in KB/TAX cells in a dose dependent manner , but had no effect on the mdr1 gene expression. These results suggest a potential action of STA-35 as MDR reversing agent, and one of the possible mechanisms could be the suppression of P-gp function and protein expression.

OBJECTIVEThe present study was undertaken to design antisense oligodeoxynucleotides (AS-ODNs) of glial glutamate transporter-la (GLT-1a) and to evaluate the effectiveness of the designed AS-ODNs on the expression of GLT-1a.

METHODSFive sequences of GLT-1a AS-ODNs were designed according to the C terminus specific sequences of GLT-1a mRNA using antisense design software of IDT Com- pany. Western blot analysis was used to evaluate the inhibition effects of the five GLT-1a AS-ODNs on the expression of GLT-la.

RESULTSThe sequence of GLT-1a AS-ODNs with sequence of 5'-GGTTCTTCCTCAACACTGCA-3' could specifically inhibit the expression of GLT-1a in the hippocampal CA1 subfield of rats, while it had no effect on the expression of GLT-1b. This sequence showed similar inhibition on the expression of GLT-la in sham and ceftriaxone (Cef)-treated rats. It could also significantly inhibit the cerebral ischemic preconditioning (CIP)-induced up-regulation in the expression of GLT-1a. The magnitude of the inhibition in sham, Cef- or CIP-treated rats was similar by more than 60%.

CONCLUSIONFrom the designed five sequences of GLT-1a AS-ODNs, we obtained an effective sequence which can specifically inhibit the expression of GLT-1a.

Animals ; CA1 Region, Hippocampal ; metabolism ; Excitatory Amino Acid Transporter 2 ; antagonists & inhibitors ; metabolism ; Ischemic Preconditioning ; Oligonucleotides, Antisense ; genetics ; RNA, Messenger ; Rats ; Up-Regulation

Withanolide A is a biologically active secondary metabolite occuring in roots and leaves of Withania somnifera. In the present study, adventitious roots from leaf explants of W. somnifera were induced for the production of withanolide-A by Agrobacterium tumefaciens strain C58C1 to obtain hair roots. Hair roots induction rate reached 30%. The withanolide A was determined by HPLC in different hair roots lines and different parts of W. somnifera. The average content of withanolide A in all hair roots lines were 1.96 times as high as that in wild-plant, the concentration of withanolide A in hair roots (1.783 mg x g(-1) dry weight) were 1.51 times as high as the roots of wild W. somnifera (1.180 mg x g(-1) dry weight), respectively. It is possible to obtain withanolide A from hair roots culture of W. somnifera.
Agrobacterium tumefaciens ; physiology ; Plant Extracts ; analysis ; biosynthesis ; Plant Roots ; chemistry ; growth & development ; metabolism ; microbiology ; Withania ; chemistry ; growth & development ; metabolism ; microbiology ; Withanolides ; analysis ; metabolism

Objective To investigate the diagnostic significance of endoscopic ultrasonography (EUS) for suspected choledocholithiasis in non-cholangiectasis. Methods EUS was performed on 33 patients with cholecystolithiasis, whose common bile duct diameters were less than 8 mm, with one of the histories of acute pancreatitis, obstructive jaundice or recurrent biliary colic, but without common bile duct stone (CBDS). The results were compared with surgical or ERCP findings. Results Twenty cases in 33 were diagnosed ascholedocholithiasis by EUS. Sixteen of the 20 cases were confirmed as CBDS with further operation or ERCP. Compared with the results of surgery or ERCP, the sensitivity, specificity, positive prediction value and negative prediction value of EUS for choledocholithiasis were 100% , 76. 5% , 80% and 100% respectively. Conclusion EUS is of high diagnostic significance for suspected choledocholithiasis in non-cholangiectasis.

OBJECTIVE:To compare genetic diversity of Spatolobi caulis from different areas of Guangxi by random amplified polymorphic DNA(RAPD)and inter-simple sequence repeat(ISSR). METHODS:Through using POPGENE 32 software,Ntsys software and SPSS 17.0 software,RAPD and ISSR methods were used to study genetic diversity of 9 samples of S. caulis from dif-ferent areas of Guangxi. RESULTS:After amplification of screened 3 RAPD primers and 4 ISSR primers,and there were 198 and 315 locus,and 37 and 80 polymorphism locus. Rates of polymorphism locus were 18.7% and 25.4%;the number of effective al-leles were 1.416 8 and 1.584 0;genetic diversity index were 0.269 4 and 0.351 3;Shannon diversity index were 0.431 6 and 0.529 9. All the values of ISSR marker were higher than RAPD marker. The average genetic similarity coefficient of ISSR and RAPD were 0.757 64 and 0.683 80,indicating ISSR was more sensitive for the detection of genetic diversity. The clustering result of them was close to each other. The correlation coefficient of them were 0.847,indicating very significant positive correlation at the level of 0.001. CONCLUSIONS:ISSR could reflect more information of genetic diversity than RAPD,and is more suitable for research of genetic diversity of S. caulis from different areas of Guangxi.

Objective To investigate the role of calcium/calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) in the primary somatosensory area (S1 area) and hippocampi in reduction of remifentanil-induced hyperalgesia by lidocaine in rats.Methods One hundred fifty-six male Sprague-Dawley rats,aged 8-10 weeks,weighing 240-260 g,were randomly allocated into 4 groups using a radom number table:control group (group C,n=6),remifentanil group (group R,n=50),lidocaine group (group L,n=50),and remifentanil+lidocaine group (group RL,n =50).Remifentanil was given as a bolus of 6 mg/kg followed by an 2 h infusion of 2.4 μg · kg-1 · min-1 in group R.Lidocaine was given as a bolus of 6 mg/kg followed by an infusion of 200 μg · kg-1 · min-1 for 2 h in group L.In group RL,drug administration was similar to those previously described in R and L groups.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before administration and at 0.5,2,5 and 24 h after the end of administration.The rats were then sacrificed immediately after administration and at 0.5,2,5 and 24 h after the end of administration in R,L and RL groups,or at the corresponding time point in group C.The S1 area and hippocampi were isolated for determination of phosphorylated CaMK Ⅱ (p-CaMK Ⅱ) expression by Western blot.Results Compared with the value before administration,the MWT was significantly decreased at 0.5 and 2 h after the end of administration (P＜0.05),and no significant change was found in TWL at each time point after the end of administration in R,L and RL groups (P＞0.05).Compared with group C,p-CaMK Ⅱ expression in the S1 area and hippocampi was significantly up-regulated immediately after administration and at 0.5 and 2 h after the end of administration in group R (P＜0.05).Compared with group R,p-CaMK Ⅱ expression in the S1 area and hippocampi was significantly down-regulated immediately after administration and at 0.5 and 2 h after the end of administration in group RL,and p-CaMK Ⅱ expression in the S1 area was significantly down-regulated immediately after administration,and at 0.5 and 2 h after the end of administration,and p-CaMK Ⅱ expression in the hippocampi was down-regulated immediately after administration,and at 0.5,2 and 24 h after the end of administration in group L,and MWT was increased at 0.5 and 2 h after the end of administration in groups L and RL (P＜0.05).There was no significant difference in TWL at each time point among the three groups (P＞0.05).Conclusion The mechanism by which lidocaine mitigates remifentanil-induced hyperalgesia is associated with inhibited activity of CaMKII in the S1 area and hippocampi of rats.

AlM: To investigate the relationship between the anterior ischemic optic neuropathy ( AlON ) and the carotid artery change using doppler ultrasound.METHODS:Fifty-four cases of AlON patients and 54 cases of healthy control were observed, atherosclerotic spots were detected by the application of color ultrasound.RESULTS:ln AlON group of 54 patients, 38 cases appeared carotid atherosclerosis, accounting for 70%. The number of cases with hard plaque, soft plaque and mixed plaques were 18, 13, and 7 respectively, accounting for 33%, 24% and 13%. ln the control group, 20 cases were detected atherosclerotic change, accounting for 37%. And the number of cases with hard plaque, soft plaque and mixed plaques were 12, 5 and 3 respectively, accounting for 22%, 9%, 6%. Significant stenosis and velocity change were showed in neither AlON group nor control group. Compared with the control group, AlON group had more cases of atherosclerotic plaque, the difference was statistically significant (χ2=12. 836, P=0. 005)CONCLUSlON: The incidence of AlON is correlated with carotid atherosclerosis, and carotid ultrasonography is significantly valuable for AlON etiology and diagnosis.

Objective To assess the technical feasibility and efficacy of the combined application of a flexible,self-expanding neurovascular stent(Neuroform)and Gugliebni detachable coils(GDC)in the management of wide-necked intracranial aneurysms in humans.Methods Sixty-five wide-necked aneurysms which underwent 65 endovascular procedures were performed by using intracranial stent and GDC.There was a total of 30 aneurysms at basilar artery including 16 at the basilar tip,9 at the basilar trunk and 5 at the beginning of the basilar artery.And there were 30 aneurysms located at the posterior communicating artery, and 5 aneurysms located at the vertebral artery.The Neuroform stents were deployed to cover the neck of aneurysms.Another microcatheter was introduced into the aneurysm sac through the stent interstices and then detachable coils were released to embolize the aneurysms.Results The combined procedures were successful in all of the 65 patients with wide-necked aneurysms.The stent could pass smoothly through the intracranial artery and got released.Complete occlusion was achieved in 60 patients and incomplete occlusion in 5 patients.In-stent thrombosis occurred in 2 patients.All patients recovered well.Forty-two patients had followe-up angiography at 3 to 6 months after the procedure.Among them,no filling was found for the 39 aneurysms which were densely packed,and 3 aneurysms had neck remnant.Conclusion The implantation of Neuroform stent as a complimentary device to GDC coiling is easy and safe for embolization of wide-necked intracranial aneurysms.It has great advantage for treatment of wide-necked intracranial aneurysms.

The complete genomes of more cyanobacterial strains have been completed for sequence, and genetic engineering of cyanobacteria has evolved in the post-genome era. Since Kaneko and colleagues had completed the sequence for the complete genome of Anabaena sp. PCC 7120 in 2001, functions of some genes in this genome, including fructose-1,6-bisphosphate aldolase (FBA) gene, have been predicated using the method of bioinformatics. However, little information is available regarding whether this gene can encode FBA and its product characteristics of related enzyme. Here, to explore this information, the predicted II-FBA gene-encoding region in Cyanobase database was cloned by PCR method and then ligated into pET-32a to generate the expression vector, pET-FBA-II. The results of SDS-PAGE indicated that the expression level of the expected target polypeptide was approximate 23.4 percent as compared to total protein and the molecular weight is about 40 kDa as compared to the protein molecular marker. The results of enzyme activity analysis showed that the activity of II-FBA was ~11.8 U per mg protein and owned a standard activity of II-FBA. To sum up, the results not only prove the functional prediction of this II-FBA gene from the Cyanobase database, but also provide the important conditions for further studying its physiological and biochemical characteristics and functions of the gene expression product.

Objective To study the mechanism of marcosomia by investigating insulin-like growth factor 2(IGF_2)imprinting status,expression level and the promoter usage in the placenta of macrosomia. Methods We selected heterozygous cases for Apa Ⅰ polymorphism in exon 9 of IGF_2 gene and then analyzed its imprinting status in 168 placentas of macrosomia and normal pregnancies.IGF_2 transcription levels and promoter usages in macrosomic and normal placenta were evaluated by using semi-quantitative RT- PCR assay.Results Thirty specimens of macrosomic placenta and 30 of normal placenta were identified as heterozygous for IGF_2.All of the heterozygous specimens showed maintenance of imprinting.The expression of placental IGF_2 mRNA(2.2?1.2)was significantly higher in macrosomia than that of normal weight group (1.6?0.6,P 0.05).Conclusion It is possible that over expression of IGF_2 in placenta contributes to macrosomia while the promoter usage and imprinting status are not associated with macrosomia.