Objective To investigate the suppression effect of RNA interference(RNAi) on the expression of epidermal growth factor receptor(EGFR) of ovarian cancer SKOV3 cell line. Methods SKOV3 cells were transfected with synthetic EGFR sequence-specific dsRNA by lipofectamin 2000.The expression level of EGFR mRNA and protein were detected by RT-PCR and Western blotting,respectively.The proliferation of transfected SKOV3 cells was detected by WST-1 cell proliferation assay.The abilities of adhesion,migration and invasion were detected by cell adhesion assay,cell migration assay and Matrigel invasion assay,respectively. Results The suppression rates on EGFR mRNA and protein by sequence-specific dsRNA-EGFR were 73.65% and 58.94%,respectively.The cell proliferation was inhibited by 22.54%,35.76% and 31.07%,respectively at 24,48 and 72 h after transfection.The adhesion of sequence-specific dsRNA transfected SKOV3 cells for 30 min and 90 min were reduced by 12.11% and 18.66%,respectively.The migration and invasion of SKOV3 cells were decreased by 25.47% and 22.08%,respectively. Conclusion The down-regulation of EGFR expression of ovarian cancer by sequence-specific dsRNA can lead to the suppression of proliferation,adhesion,migration and invasion of ovarian cancer cells.

Nerve segments were excised from the posterior limb of the adult dog and treated by repeated freezing and thawing for five times. A segment about 5 mm long was removed from the right tibial nerve proximal to the popliteal fossa of each experimental adult rabbit, and 8 mm of nerve segment from the dog prepared as above was transplanted in the gap. Both the proximal and distal ends of the tibial nerve of host rabbits were sutured to the graft nerve of the donor dog. The grafts were excised together with the sutured cut ends of the recipient tibial nerve at 3, 4, 5, 6, 7, 15, 18, and 31 weeks after transplantation. The regenerating nerve fibers were found by light microscope from the proximal end of the right tibial nerve to the graft, and from the graft to the distal part of the tibial nerve at 3, 4, 5, 6, 7, 15, 18 and 31 weeks. Under electron microscope regenerating myelinated and unmyelinated fibers were found to be separate or in fascicle in the graft and the distal segment of the tibial nerve at the above survival time after transplantation. Perineurium was seen surrounding the regenerating nerve fascicles. Neurotubules, neurofilaments and mitochondria were found in the regenerating axons. The diameter of the regenerating myelinated fibers with long survival time after transplatation was thicker than that with short survival time. Repeated freezing and thawing reduced the antigenicity of the heterogenic nerve that it was not rejected after transplantation, and induced the regenerating nerve fibers of the host grow into the graft nerve and extend distally.

The effect of concanavalin A (Con A) on nerve regeneration in heterograft was done in the present study. A segment about 5mm long was removed from the right sciatic nerve of adult rats. A 8mm of the heterograft nerve segment from the tibial nerve of the rabbit and pretreated by Con A was transplanted into the gap of the severed rat nerve. Regenerating nerve fibers were found from the proximal part of the sciatic nerve into the graft, and also from the graft into the distal part of the sciatic nerve at the 4th,8th and 12th week after transplantation. Regenerating nerve fibers existed separately or clustered into fascicles in the graft,as well as in the distal part of sciatic nerve. Some unmyeli-nated nerve fibers were scattered among the regenerating myelinated fibers. The gastrocnemius muscle was AChE-positive at 8th and 12th week after transplantation. In silver-combined AChE staining section, the regenerating nerve fibers could be seen connecting with the motor endplates. There were regenerating free nerve endings and nerve fascicles in the dermis and epidermis at 8th and 12th week after transplantation. The present study showed that the pretreatment of Con A had benefit to that the regenerating nerve fibers to pass through the heterograft and reinnervated to the targetes.ADepartment of Anatomy,Jiangxi Medical College,Nanchang 330006,China

Objective To observe the change of six cytokines in mice infected with Echinococcus multilocularis as part of the study on immunological mechanism in the infection. Methods Mice were infected by abdominal inoculation of echinococcus protoscoleces. The change of serum level of the cytokines IL-2、IFN-?、TNF-?、IL-4、 IL-5 and IL-10 was determined by ELISA during the infection which lasted for 260 d. Results Compared with uninfected control, the levels of the cytokines all significantly increased in the 260 d. The level of IL-2 reached a peak after 80 d post-infection (p.i.), then decreased quickly after 140 d p.i., High level of TNF-? was detected after 40 d, compared to uninfected control, reached a peak at 100 d p.i., and decreased quickly after 140 d. The level of IFN-? reached a peak after 80 d p.i., and decreased slowly after 140 d p.i., The levels of IL-4, IL-5 and IL-10 remained lower before 80 d, and increased sharply after 100 days. The levels of IL-4 and IL-10 reached peaks at 100 d p.i., and that of IL-5 at 140 d p.i. Conclusion The data suggest that the induction of Th2 antibody-mediated immunity (AMI) with a parallel expansion of Th1 cell-mediated inflammatory (CMI) responses are important mechanism of the host in defending against the metacestodes. Th1 CMI plays an important role at the early stage of infection, and Th2 AMI is important in the later stage of infection.

Objective:Both URL and MPCNL have been widely applied in the treatment of ureteral calculi.In this study,the safety,efficiency and degree of injury were compared between retrograde and antegrade access ureterolithotripsy for the calculi complicated with infection,in order to clarify the indications of the two types of minimally invasive techniques.Methods:A total of 192 patients with upper ureteral calculi complicated with infection were treated with ureterolithotripsy.The operation was performed via retrograde transurethral access in 72 patients and via antegrade percutaneous nephrostomy tract in 120 patients.Results:The success rate of retrograde approach was 93.1%.Mean operating time was 45.9?16.5min(20-90 min) with mean hospital stay of 5.1?1.8(2-6) days.The stone free rate was 84.7%(61/72) at 1 month follow-up.5 patients with residual calculi required combined ESWL.Complications were noted in 5.6%,CRP was 10.59?5.12 mg/L before operation,and was 38.63?4.61 mg/L 24h after the operation.The success rate of antegrade approach was 99.2%.Mean operating time was 62.4?17.6 min(40-120 min) with mean hospital stay of 8.57?2.57(5-15) days.The stone free rate was 100% and 1 complication was noted(0.01%).CRP was 11.29?5.38 mg/L before the operation,34.93?7.82 mg/L 24h after the operation.For the success rate and stone free rate,antegrade approach was higher than retrograde approach(P0.05)between the two groups before operation,but antegrade approach had lower CRP(P

The study is to develop a method to determine 3 batches leaves of Nauclea officinalis and stems of N. officinalis by HPLC. The differences between strictosamide contents and fingerprints was compared, then chromatographic peak of fingerprints was validated with the assistance of LC-MS. The strictosamide contents in stems of N. officinalis were higher than leaves of N. officinalis. The main chemical composition in leaves of N. officinalis and stems of N. officinalis were alkaloid which revealed by LC-MS. There are 7 chemical compositions were same between them, but the chemical composition in leaves of N. officinalis is more than stems of N. officinalis. This provides a scientific basis for the development of the potential medicinal value of leaves of N. officinalis and the sustainable utilization of N. officinalis.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Rubiaceae ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Two endophytic-bacteria isolates of G18 and F19 were isolated from the stem of Taxus chinensis. The G18 and F19 were respectively classified into Psudomonas sp. and Stenotrophomonas sp. based on biological characteristics and 16S rDNA sequence analysis. The bioactivity analysis showed that the fermented broths of the G18 and F19 exhibited antagonistic activities against three pathogenic bacteria, and had good antagonistic effectiveness to Verticillium dahliae and Colletotrichum gloeosporioides, respectively. The G18 can degrade salicylic acid, and the F19 can do dichlorvos.

AIM To explore pharmacokinetics of Berberine in YL2000 in normal and febrile rats. METHODS The levels of Berberine in plasma were measured through HPLC and secondary parameters were obtained by fitting the dose time data of Berberine making use of 3P87 programme. RESULTS In normal and febrile rats, the plasma concentration of Berberine was peaked at (3 4?0 3) h vs (0 3?2 1) h( P

Objective To compare the normal anatomy of the anterior cruciate ligament (ACL) of fresh frozen cadaveric knee specimen in oblique coronal thin-slice section with oblique coronal magnetic resonance imaging. Methods One fresh cadaveric knee specimen was scanned with MR T1-weighted spinecho sequence.then the specimen was frozen and sliced with a band saw along the oblique coronal plane into 1.0-mm-thick sections that corresponded to the MR images,MR images including oblique coronal T1-weighted and T2-weighted images of 50 normal the knee joints were retrospectively reviewed to observe the MR imaging features of the cruciate ligament. Results Anteromedial and posterolateral bundles of ACL were clearly depicted on both anatomic slices and MR images.The anteromedial bundles originated from the posteromedial aspect of the lateral femoral condyle,coursing through the lateral intercondylar notch in an anterior,inferior,and medial direction,and inserted on the anteromedial aspect of the intercondylar eminence. The posterolateral bundles originated from the anteromedial aspect of the lateral femoral condyle,passing laterally and inferiorly through the lateral intercondylar notch,and inserted on the posterolateral side of the intercondylar eminence.The full length of ACL of all 50 individuals was showed on MR images.MRI clearly differenitated the anteromedial and posterolateral bundles of ACL and depicted the full length of the bundles.similar to the findings on sectional anatomy.Conclusion Oblique coronal MR imaging is the best way to demonstrate ACL and should be used for clinically suspected injury of ACL.

OBJECTIVE:To establish the method for the determination of mildronate in human plasma and urine,and to study the pharmacokinetic characteristics in healthy volunteers. METHODS:After precipitating plasma and urine sample,LC-MS/MS method was adopted. Dikma Diamonsil C18 column was used with mobile phase consisted of methanol-water(containing 0.2% for-mic acid,0.3% ammonium acetate)(31∶69,V/V)at the flow rate of 0.6 ml/min. ESI was adopted in MRM mode,by using nega-tive ion. The ion for quantitative analysis were m/z 147.10→58.20 (mildronate) and m/z 152.00→110.10 (internal standard,acet-aminophen). The pharmacokinetic parameters of mildronate with single administration and multiple administration were calculated by using DAS 2.1 software and compared. RESULTS:The linear range of mildronate in plasma were 0.02-20 ng/ml(r=0.999 3) and in urine were 0.05-40 ng/ml(r=0.998 2). The lowest limits of quantitation were 0.02 and 0.05 ng/ml. Precision and recovery met the requirements of biological specimen determination,and endogenous impurities hadn’t effect on the determination. The main pharmacokinetics parameters of low-dose,medium-dose and low-dose(250,500,750 mg)of mildronate in plasma with single ad-ministration were as follows:t1/2 were(3.39±0.81),(5.52±0.57)and(5.32±0.96)h;tmax were(0.80±0.45),(1.38±0.43)and (1.10±0.36)h;cmax were(4.17±1.46),(8.08±1.04)and(15.04±1.86)ng/ml;AUC0-36 h were(24.55±5.81),(45.50±7.07)and (85.60 ± 13.09)ng·h/ml. In the dose range,cmax,AUC0-36 h h had a linear relationship with dose (R2 were 0.974 5 and 0.968 3). The main pharmacokinetic parameters of low-dose of mildronate with multiple administration after keeping stable were as follows:cmin was(0.28 ± 0.10)ng/ml;AUCs was(38.78 ± 4.18)ng·h/ml;cs was(1.62 ± 0.17)ng/ml;DF was(3.81 ± 1.14);t1/2 was(6.17 ± 1.46)h;tmax was(1.20 ± 0.33)h;cmax was(6.46 ± 1.96)ng/ml;AUC0-36 h was(40.33 ± 4.65)ng·h/ml;accumulation factor of cmax and AUC were(1.73±0.90)and(1.64±0.40). Compared with single administration,t1/2,cmax and AUC of mildronate with multiple admin-istration after keeping stable all changed,and tmax had no signifi-cant difference. After single administration,26 h accumulative excretion rate of those groups were (0.004 009 ± 0.001 1)%, (0.004 026±0.001 01)% and(0.003 858±0.000 68)% respec-tively. CONCLUSIONS:Established method is sensitive,accurate and specific,and suitable for the determination of mildronate concentration in human plasma and urine and pharmacokinetics study. Mildronate capsule shows certain accumulation effect in healthy volunteers,and linear pharmacokinetic characteristics.