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.

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

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.

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

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

Objective: To observe the regulatory effects of miRNA-31 (miR-31) on LATS2 and cardiomyocyte hypertrophy via down-regulating miR-31 expression in rat's cardiomyocytesin vitro. Methods: Rat's cardiomyocytes were isolated and cultured for 10 daysin vitro, according to different intervention methods, the cells were divided into 4 groups:①Blank control group,②AngII intervention group,③Lentivirus with miR-31 inhibitor infection group,④Negative lentivirus infection group. On day-8, gene expressions of MiR-31, LATS2, cardiac hypertrophy ANP and β-MHC were examined by qRT-PCR; on day-10, cell morphology was observed by fluorescence staining. LATS2 protein expression was examined by Western blot analysis. Dual luciferase reporter plasmids were transfected into 293T cells, then luciferase activity was detected to identify the targeting effect of miR-31 on LATS2. Results: Compared with Blank control group, AngII intervention group showed increased gene expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, enlarged cardiomyocyte surface,P<0.05; while decreased gene and proteinexpressions of LATS2,P<0.05. Compared with AngII intervention group, Lentivirus with miR-31 inhibitor infection group had down-regulated expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, reduced cardiomyocyte surface, P<0.05; while slightly increased LATS2 gene expression and obviously increased protein expression,P<0.05. Dual luciferase reporter assay presented that relative luciferase activity of TRAF6-3' UTR+miR-146b was significantly decreased than TRAF6-3' UTR+miR-NC,P<0.01 and relative luciferase activity of LATS2-3' UTR+ miR-31 was signiifcantly reduced than LATS2-3' UTR-NC+miR-31,P<0.01. Conclusion: Cardiomyocytes hypertrophy could be reversed at certain degree by down-regulating miR-31; the targeting effect of miR-31 on LATS2 was involved in cardiomyocyte hypertrophyregulation.

OBJECTIVETo investigate the protective effects of cistanche total glycosides (CTG) on dopaminergic neuron in substantia nigra (SN) of model mice of Parkinson's disease (PD).

METHODSExperimental mice were randomly divided into 5 groups, the normal control group, the model group, the high (400 mg/kg), moderate (200 mg/kg) and low (100 mg/kg) dose CTG groups. Mouse model of chronic PD was induced by peritoneal injection of MPTP (1-methyl-4-phenyl-1,2,3,6-ttrahydropyridine) 30 mg/kg for 5 successive days. Climbing test was used to estimate the neurobehavior of mice on the 7th and 14th day (D7 and D14) after initiating MPTP injection; meantime, quantitative immunohistochemistry was conducted to detect the number of dopaminergic neuron in SN and expression of tyrosine hydroxylase (TH) in striatum.

RESULTSThe average time of climbing in the high dose CTG group on D7 and D14 was significantly shorter than that in the model group (P < 0.01). The mean optic density (OD) of TH in striatum was higher in the three CTG groups than that in the model group on D7 (P < 0.01); but on D14, significance only showed in the high and moderate dose CTG groups (P < 0.01). Moreover, the MPTP induced decrease of TH positive neuron could be antagonized by CTG, but significant difference only showed between the high dose CTG group and the model group at the two time points of observation (P < 0.05).

CONCLUSIONCTG could improve the neurobehavior of PD model mice significantly, and inhibit the decrease of nigral dopaminergic neurons and TH expression in striatum.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ; Animals ; Behavior, Animal ; drug effects ; Cistanche ; chemistry ; Dopamine ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Glycosides ; pharmacology ; Immunohistochemistry ; Male ; Mice ; Mice, Inbred C57BL ; Neurons ; drug effects ; metabolism ; pathology ; Neuroprotective Agents ; pharmacology ; Parkinson Disease, Secondary ; chemically induced ; physiopathology ; Random Allocation ; Substantia Nigra ; drug effects ; metabolism ; pathology ; Tyrosine 3-Monooxygenase ; metabolism

OBJECTIVETo study the differences in the toxicity of vinegar-processed Kansui Radix on normal and cancerous ascites model rats.

METHODNormal and cancerous ascites model rats were taken as the research objects and orally administered with different doses of vinegar-processed Kansui Radix for 7 d. Pathological sections were prepared to observe the damages in liver, stomach, intestinal tissues in rats and detect the impacts on serum, liver, stomach and intestinal tissues and the oxidative damage index.

RESULTCompared with the blank group, all of normal administration groups and model groups showed significant damages in liver, stomach and intestinal tissues. Compared with the model groups, all of normal administration groups revealed notable alleviation in damages. Compared with the blank group, the model groups showed significant increases in AST, ALT and MDA in serum and liver (P < 0.01) and a significant decrease in GSH in serum and liver, stomach, intestinal tissues (P < 0.01). Compared with the blank group, the results showed significant decreases in ALT, AST in serum and ALT in liver in model low, medium and high dose groups and AST activity in liver tissues in the normal high dose group (P < 0.05, P < 0.01); significant decreases in GSH in serum and stomach tissues in normal low, medium and high dose groups and GSH content in liver and intestinal tissues in normal medium and high dose groups (P < 0.05, P < 0.01); notable rises in MDA in liver tissues in normal low, medium and high dose groups and MDA content in serum and stomach and intestinal tissues in normal medium and high dose groups (P < 0.05, P < 0.01). Compared with model groups, data revealed significant decreases in ALT, AST in serum in model low, medium and high dose groups, AST in liver tissues of model medium and high dose groups and ALT activity in liver in the model high dose group (P < 0.05, P < 0.01); significant increases in GSH content in serum and stomach tissues of model low, medium and high dose groups, GSH in liver tissues in model medium and high dose groups and GSH in intestinal tissues in the high dose groups (P < 0.05, P < 0.01); and notable declines in MDA content in serum in model low, medium and high dose groups, MDA in liver tissues of model medium and high dose groups and MDA in stomach and intestinal tissues the high dose group (P < 0.05, P < 0.01).

CONCLUSIONAccording to the study, vinegar-processed Kansui Radix showed a significant lower toxicity liver, stomach, and intestines of cancerous ascites model rats, which provided a basis for clinical safe application of vinegar-processed Kansui Radix based on symptom-based prescription theory.

Acetic Acid ; chemistry ; Animals ; Chemistry, Pharmaceutical ; methods ; Drug Prescriptions ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; toxicity ; Euphorbia ; chemistry ; toxicity ; Intestines ; drug effects ; pathology ; Liver ; drug effects ; metabolism ; pathology ; Male ; Neoplasms ; drug therapy ; metabolism ; pathology ; Oxidative Stress ; drug effects ; Plant Roots ; chemistry ; toxicity ; Rats ; Rats, Sprague-Dawley

Limit test of flavones in diterpene ginkgolides meglumine injection materials by UV-Vis and HPLC-DAD method was studied in this essay. The HPLC-DAD method has lower LOD (about 1% of the UV-Vis), that is, the sensitivity is higher than UV-Vis method. Through the analysis of the kinds of flavonoids ingredients in the samples by LC-MS, the three compounds with highest contents are kaempferol, quercetin and isorhamnetin. Kaempferol, quercetin and isorhamnetin were chosen as reference compounds for HPLC analysis, and the HPLC separation analysis was carried on an Agilent Eclipse plus C18 column (4.6 mm x 250 mm, 5 μm) with methanol and water containing 0.4% phosphoric acid (50: 50) as mobile phase, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. This method has good specificity, precision and reproducibility. The LODs of quercetin, kaempferide and isorhamnetin were 27.6, 22.3, 29.5 μg x L(-1). The average recovery was 87.9% (RSD 3.3%), 91.7% (RSD 3.1%), 88.3 (RSD 1.3%) for quercetin, kaempferide and isorhamnetin, respectively. Based on the 10 batches of sample results and sensitivity of different HPLC, the content of total flavonoids ingredients of diterpene ginkgolides meglumine injection materials was limited no more than 2 x 10(-5). This method is simple, quick and has good maneuverability, and could be used to the limit test of flavonoids in the diterpene ginkgolides meglumine injection materials.
Chromatography, High Pressure Liquid ; methods ; Diterpenes ; analysis ; Drugs, Chinese Herbal ; analysis ; Flavones ; analysis ; Ginkgolides ; analysis ; Limit of Detection ; Mass Spectrometry ; methods