Objective To summarize the treatment results,operative complications and experiences of laparoscopic modified pull-through(Soave)precedure.Methods Twenty children with Hirschsprung disease(HD),16 males and 4 females,with age from 3.5 months to 4 years old,underwent laparoscopic modified pull-through(Soave)precedure under whole anesthesia and tracheal intubation,and the pathological colon was freed by ultrasonic surgical knife.Results All cases were operated on successfully.The average operating time was 132 min.The average resection colon was 35 cm and the average volume of bleeding was about 14 mL without blood transfusion.Two patients had postoperative enterocolitis;3 patients were no complications postoperation,such as constipation,soiling,intestinal obstruction,preianal intestine prolapse or cuff narrowing,and all patients had 1-2 bowel movements per day during a period of 6 months to 3 years follow-up.Conclusion Laparoscopic modified pull-through(Soave)precedure has smaller incision,less bleeding,quick recovery and less complications.

Ginsenosides are the abundant secondary metabolites in American ginseng (Panax quinquefolium), it could be released into soil through root exudation and decomposition during plant growth. This study determined ginsenoside contents in American ginseng cultivated soil by HPLC. Three ginsenosides, Rb1, Rb2 and Rd, were detected in the rhizosphere soil of 3-4 years old American ginseng cultivated in Huairou District, Beijing, and their contents were 0.80-3.19 mg x kg(-1). Correspondingly, the contents of the three ginsenosides in soil solution were 4-16 mg x L(-1) at field water-holding capacity of 20%. According to the field soil test data, we designed the concentration of ginsenosides for bioassays (0.2-125 mg x L(-1) in solution or 0.2-125 mg x kg(-1) in soil). The results showed that radicle lengths of American ginseng were reduced by 6%-23% in solution containing 0.2-125 mg x L(-1) ginsenoside extract, and a significant difference was observed at concentration of 125 mg x L(-1) (P < 0.05). The shoot lengths of American ginseng were not significantly inhibited by 0.2-125 mg x L(-1) ginsenosides extractions. After 20 days of growth in nutrient solution amended with 25 mg x L(-1) ginsenosides extraction, plant height of 3-year-old American ginseng seedling was decreased by 28% compared to the control, and the biomass of aerial parts was also reduced by 50% (P < 0.05). However, the growth of newly-grown fibrous root was not significantly inhibited. Comparatively, when American ginseng embryos were cultivated into sterile or non-sterile soil, neither radicle lengths nor shoot lengths were significantly affected by 0.2-125 mg x kg(-1) ginsenoside extracts. In conclusion, ginsenosides showed autotoxic effect on growth of American ginseng radicle and adult seedling, however, this effect was weakened in field soil.
Chromatography, High Pressure Liquid ; Culture Media ; chemistry ; metabolism ; Ginsenosides ; analysis ; metabolism ; toxicity ; Panax ; chemistry ; drug effects ; growth & development ; metabolism ; Plant Roots ; chemistry ; drug effects ; growth & development ; metabolism ; Soil ; chemistry

OBJECTIVETo explore the difference in the efficacy and effect mechanism of subcortical ischemic vascular disease (SIVD) complicated with depression between acupuncture and medication.

METHODSSixty patients were randomized-into an acupuncture group and a medication group, 30 cases in each one. In the acupuncture group, acupuncture was applied to Baihui (GV 20), Shuigou (GV 26), Fengfu (GV 16), Fengchi (GB 20) and the others, once a day, 6 times a week. The treatment of 4 weeks made one session and totally 2 sessions were required. In the medication group, nimodipine 30 mg, three times a day and fluoxetine 20 mg, once a day were prescribed for oral administration, for 8 weeks totally. Before treatment, at the end of the 4th week and at the end of the 8th week of treatment, cerebral blood flow velocity (CBFV) and solubility CD40 ligand (sCD40L) were determined respectively. The scores in Montreal cognitive assessment (MoCA) and Hamilton' s depression scale (HAMD) were evaluated in the two groups. The efficacies on cognitive function and depression symptoms were compared in the patients between the two groups. Results Compared with the outcome before treatment, mean blood flow velocity (Vm) of middle cerebral artery (MCA), anterior cerebral artery (ACA) and posterior cerebral artery (PCA) was increased significantly at the end of the 4th week of treatment in the two groups (all P<0.05). At the end of the 8th week, Vm was increased much significantly (all P<0.01). The differences were not significant in comparison between the two groups (all P>0.05). Compared with the expression before treatment, sCD40L was reduced significantly after treatment in the patients of the two groups (all P<0.01), but the differ- ence was not significant between the two groups (all P>0.05). Compared with that before treatment, MoCA score was increased significantly after treatment in the two groups (all P<0.01), HAMD score was reduced sig- nificantly (all P<0.01), the differences were not significant in comparison between the two groups (all P>0.05). The total effective rate of cognitive improvement was 86.7% (26/30) in the acupuncture group and was 80.0% (24/30) in the medication group, the differences were not significant in comparison of the two groups (P>0.05). The total effective rate of the improvement in depression was 93.3% (21/30) in the acupuncture group and was 86.7% (26/30) in the medication group, the differences were not significant in comparison of the two groups (P>0.05).

CONCLUSIONAcupuncture could significantly increases CBFV and reduces serum sCD40L expressions in the patients of SIVD complicated with depression, and significantly improves cognitive function and relieves depression symptoms. The efficacy of it is similar to that of western medication. The increase of serum sCD40L expression is possibly involved in the occurrence and development of SIVD. Reducing sCD40L expression contributes to the alleviation of damage induced by cerebral ischemia and reperfusion.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Cerebral Infarction ; complications ; drug therapy ; therapy ; Depression ; drug therapy ; etiology ; therapy ; Female ; Fluoxetine ; administration & dosage ; Humans ; Male ; Middle Aged ; Nimodipine ; administration & dosage ; Treatment Outcome

Animals ; Electromagnetic Fields ; Embryo, Mammalian ; metabolism ; Female ; Mice ; Pregnancy ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

The present study is to elucidate the mechanisms underlying Gleevec-induced apoptosis of chronic myeloid leukemia (CML) K562 cells in vitro. The apoptotic cell death and cell cycle distribution after Gleevec treatment and the effect of PDCD4 siRNA on Gleevec-induced apoptosis of K562 cells were analyzed by flow cytometry. The effect of Gleevec on p-Crkl, caspase-3, PARP and PDCD4 protein levels, and the knockdown efficacy of PDCD4 siRNA were detected by Western blotting. The results showed that Gleevec dramatically suppressed the phosphorylation level of Crkl in a dose-dependent manner and induced significant apoptosis and G0/G1 cell cycle arrest of K562 cells in time- and dose-dependent manners. In addition, Gleevec activated caspase-3 and its downstream substrates PARP, and the caspase pan inhibitor Z-VAD-FMK (50 micromol x L(-1)) markedly reduced Gleevec-induced apoptosis from 47.97% +/- 10.56% to 31.05% +/- 9.206% (P < 0.05). Moreover, Gleevec significantly increased the protein expression of programmed cell death 4 (PDCD4). PDCD4 knockdown by siRNA reduced Gleevec-induced apoptosis from 46.97% +/- 14.32% to 42.8% +/- 11.43%. In summary, Gleevec induced apoptosis in K562 cells via caspase-3 activation.
Amino Acid Chloromethyl Ketones ; Apoptosis ; drug effects ; Benzamides ; pharmacology ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Humans ; Imatinib Mesylate ; K562 Cells ; Phosphorylation ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology

OBJECTIVETo explore the effect of Jingui Shenqi pill (JGSQP) with various concentrations at different time points on pituitary adrencorticotropic hormone (ACTH) gene expression level in Shen-Yang deficiency rats.

METHODSThe Shen-Yang deficiency rats were randomly divided into the model control group and the high, medium and low dosage of JGSQP groups. Reverse transcriptase polymerase chain reaction was used to observe the effect of JGSQP on the ACTH mRNA of pituitary tissue in rats treated at different time points (10 d, 20 d, 30 d).

RESULTSAs compared with that in the model group, the ACTH gene expression level was significantly higher in the high dose JGSQP group (P < 0.05), and the increment in the medium dosage group was significantly higher in comparing with that in the high and low dosage groups (P < 0.05 or P < 0.01).

CONCLUSIONThrough up-regulation on ACTH gene expression is possibly one of the mechanisms of JGSQP in treating Shen-Yang deficiency.

Adrenocorticotropic Hormone ; biosynthesis ; genetics ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Hypothalamo-Hypophyseal System ; Kidney Diseases ; genetics ; metabolism ; Medicine, Chinese Traditional ; Pituitary Gland ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Yang Deficiency ; genetics ; metabolism

Fourteen compoumds were isolated from the ethyl acetate portion of the 95% ethanolic extract of Pleione bulbocodioides by a combination of various chromatographic techniques including silica gel, ODS, macroporous adsorbent resin, Sephadex LH-20, and preparative HPLC, of which ten compoumds were phenanthrenes and dihydrophenanthrenes, two compoumds were bibenzyls, one was lignan and a sterol. Their structures were identified on the basis of spectroscopic data as monbarbatain A(1), 2, 7, 2'-trihy-droxy-4, 4', 7'-trimethoxy-1, 1'- biphenanthrene(2), blestriarene A(3), pleionesin B(4), shanciol H(5), 17-hydroxy-7'-(4'-hy-droxy-3 '-methoxyphenyl)- 4-methoxy-9, 10, 7', 8'-tetrahydrophenanthro[2, 3-b]furan-8'-yl methyl acetate(6), 1-p-hydroxybenzyl-4-methoxy phenanthrene-2, 7-diol(7), 1-p-hydroxybenzyl-4-met-hoxy-9, 10-dihydrophenanthrene-2, 7-diol(8), hircinol(9), coelonin( 10), gigantol(11), batatasin 11 (12), syringaresinol(13) and ergosta4, 6, 8 ( 14) , 22-tetraen-3-one (14). Compounds 1-3, 9, 13 and 14 were isolated from this genus for the first time.
Drugs, Chinese Herbal ; chemistry ; Orchidaceae ; chemistry ; Organic Chemicals ; analysis

OBJECTIVETo observe the impact of bone morphogenetic protein-2 (rhBMP-2) on bone marrow stromal cells (BMSCs) osteogenesis in vitro and vascular endothelial growth factors (VEGF) expression in bone osteoporotic to prevent and treat the osteoporosis.

METHODSTwenty 6-month-old female SD rats weighted (300±20) g underwent bilateral ovariectomized. At 3 months after operation, dual-energy X-ray absorptiometry was used to measure bone mineral density of rats,the values were compared with preoperative to ensure the model successfully, and the osteoporosis rats' BMSCs were cultured by bone marrow adherent cultured and the BMSCs morphology was observed under a phase contrast microscope upside down. The osteoporosis rats' BMSCs at the 2nd generation (p2) were randomly divided into experimental and control groups and were added complete medium (containing rhBMP-2) and osteogenic induced liquid, respectively. Two weeks later, the formation of cell calcium nodules were detected by Alizarin red staining method,alkaline phosphatase activity was measured by enzyme standard instrument and the expression of VEGF was detected by RT-PCT method.

RESULTS(1)Whole body bone mineral density of rats before and after operation were (0.179±0.007), (0.158±0.006) g/cm2,there was statistically significant (t=4.180,P< 0.05). (2)Alizarin red staining at 2 weeks after osteogenesis induced by BMSCs (P2) in the experimental group had more strong dyeing effect than the control group obviously. (3)Alkaline phosphatase activity at 2 weeks after osteogenesis induced by BMSCs (P2) of the experimental group (15.62±1.27) ug/gprot was significantly higher than that of the control group (8.62±0.93) ug/gprot,there was statistically significant (t=7.709, P<0.01). (4)The expression of VEGF at 2 weeks after osteogenesis induced by BMSCs (P2) of the experimental group 3.723±0.143 was significantly higher than that of the control group 0.950±0.072, there was statistically significant (t=29.462, P<0.01).

CONCLUSIONRhBMP-2 can improve the in-vitro osteogenesis ability of ovary osteoporosis rat BMSCs, promote the VEGF expression of osteogenesis factor. Regulating the VEGF expression may be one of the mechanisms of BMP-2 to participate in bone metabolism.

Animals ; Bone Density ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Cells, Cultured ; Female ; Humans ; In Vitro Techniques ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Osteogenesis ; Osteoporosis, Postmenopausal ; genetics ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Epithelial-to-mesenchymal transition (EMT) plays a critical role in cancer metastasis,and is relevant to the inflammatory microenvironment.Lipopolysaccharide (LPS),a cell wall constituent of gram-negative bacteria,has been reported to induce EMT of cancer cells through TLR4 signal.We previously reported that LPS promoted metastasis of mesenchymallike breast cancer cells with high expression of cyclin D 1 b.However,the role of cyclin D1b in LPS-induced EMT has not been fully elucidated.In the present study,we described that cyclin D1b augmented EMT induced by LPS in MCF-7 breast cancer cells.Cyclin D1b markedly amplified integrin αvβ3 expression,which was further up-regulated under LPS stimulation.Our results showed ectopic expression of cyclin D1b promoted invasiveness of epithelial-like MCF-7 cells under LPS stimulation.Additionally,LPS-induced metastasis and EMT in MCF-7-D1b cells might depend on αvβ3 expression.Further exploration indicated that cyclin D1b cooperated with HoxD3,a transcription factor promoting αvβ3 expression,to promote LPS-induced EMT.Knockout of HoxD3 repressed LPS-induced EMT and αvβ3 over-expression in MCF-7 cells with high expression of cyclin D1b.Specifically,all these effects were in a cyclin D1a independent manner.Taken all together,LPS up-regulated integrin αvβ3 expression in MCF-7 cells with high expression of cyclin D 1b and induced EMT in breast cancer cells,which highlights that cyclin D1b may act as an endogenous pathway participating in exogenous signal inducing EMT in breast cancer cells.

Background There are a lot of studies about the carrier of corneal endothelial transplantation,but the best carrier has not been defined.Objective This study was to investigate the biocompatibility of chitosan carrier with rabbit corneal endothelium in vivo.Methods Fresh eye-balls were obtained from 10 New Zealand white rabbits.Rabbit corneal endothelial cells (CECs) were isolated and cultured on chitosan carrier in vitro.The morphology and density of rabbits CECs were observed every day,and the expressions of fibronectin (FN),collagen-1 (Coil-I) and Zonula occludens 1 (ZO-1) were detected by immunoinfluorescence.The morphology and ultrastructure of CECs were observed under the scanning and transmission electron microscope.Chitosan carrier with CECs was implanted into the anterior chamber of the left eyes in ten healthy New Zealand white rabbits,and only paracentesis of anterior chamber was performed in the right eyes as controls.The inflammation of ocular anterior segment was examined under the slit lamp microscope,and corneal thickness was measured 1 week,4 and 8 weeks after operation.Corneal endothelium cell density and morphology were examined under the corneal endothelial microscope at postoperative 2 weeks.Corneal samples were collected for the regular histopathological examination to observe the inflammatory reaction at postoperative 1 month and 3 months.Paired t test was used for statistical analyses between the control group (left eyes) and the experimental group (right eyes).The use and care of the animals followed the Statement of ARVO.Results CECs formed an intact monolayer of cells with the uniform shape and size on the chitosan membrane after incubated for 5 days.The cells reached confluence of 90％ 7 days after cultured with the 40％ hexagon cells.Under the scanning electron nicroscope,rabbit CECs showed the round or polygon in the shape with the microvillus on the cell surface.The cells connected closely by desmosome.The processes,pseudopodiums and microvillus on the cellular surface,vacuole in the cytoplasm,expanded endoplasmic reticulum with ribosome and abundant chromatin were exhibited under the transmission electron microscope.The immunofluorescence examination revealed the positive expressions of FN,Coll-Ⅰ and ZO-1 in the CECs on the chitosan carrier.In the in vivo experiment,the exudation in the anterior chamber and corneal edema were seen under the slit lamp microscope 3 days after implantation of chitosan carrier with CECs.However,the inflammation was gone 14 days after operation.The differences of the corneal thickness were no significant between the experimental group and the control group 1 week and 4,8 weeks after operation (t =1.377,P=0.265;t =1.795,P=0.165 ; t =0.390,P =0.760).In addition,no significant differences were found in the CECs density and the hexagon cells rate between the two groups(P =0.365,0.062).The histopathological examination showed that the inflammatory cells around the chitosan membrane were disappeared 3 months after operation and showed a good corneal structure.Conclusions Chitosan carrier has a good biocompatibility with rabbit CECs and anterior chamber,and it may be a potentially good carrier for CECs transplantation.