Objective To explore the efficacy and safety of mild hypothermia (MH) in treating the infants with moderate-to-severe neonatal hypoxic-ischemic encephalopathy(HIE),and to make a follow-up of the nerve motor development of the infants at 18 months old after discharge.Methods Totally 61 neonates with moderate-to-severe HIE in Neonatal Intensive Care Unit (NICU) from Jan.2007 to Dec.2013 were retrospectively analyzed.According to before and after MH therapeutic apparatus was used by NICU of Shanghai Children's Hospital,61 neonates of HIE were divided into 2 groups,the conventional treatment group(25 cases) and MH treatment group(36 cases).The patients in both groups were measured respectively by using the amplitude integrated electroencephalography (aEEG) before MH treatment and at 72 hours after M H treatment,by neonatal behavioral neurological assessment(NBNA) on the 28th day after birth,and by adopting Bayley Scales of Infant Development at 18 months old.The adverse reactions,serious disability cases and deaths of MH treatment were recorded.Results Compared with the conventional treatment group,aEEG recording before treatment showed no statistically significant differences in MH treatment group [maximum voltage:(22.4 ±3.1) μV vs(18.6 ±2.5) μV,maximum voltage:(8.2 ±2.6)μV vs(6.5 ±1.9) μV,t =1.264,0.852,all P ＞ 0.05].However,aEEG recording at 72 h after treatment showed statistically significant differences in MH treatment group [maximum voltage:(24.1 ± 3.2) μV vs (30.6 ± 2.8) μV,maximum voltage:(9.7 ± 3.4) μV vs (13.3 ± 2.2) μV,t =6.376,4.257,all P ＜ 0.05].Severe disability cases [24.0％ (6/25 cases) vs 5.6％ (2/36 cases),x2 =4.405,P ＜ 0.05] and deaths [16.0％ (4/25 cases) vs 0 (0/36 case),x2 =6.1 64,P ＜ 0.05] in MH treatment group were significantly decreased,and there was significantly difference in NBNA on the 28th day after birth[(35.9 ± 2.1) vs(39.1-± 1.6),t =3.361,P ＜ 0.05],and scales of neurobehavioral evaluation through follow-up of 18 months old [mental development index (MDI):(85.2 ± 10.7) vs (96.5-± 13.1),t =7.839,P ＜ 0.05].Very few neonates had apnea,coagulation dysfunction,arrhythmia and other adverse reactions in MH treatment course.Conclusions MH treating moderate-to-severe HIE is safe and effective.MH is effective in reducing death and major disabilities in neonates with moderate-to-severe HIE and without significant side effects.MH can obviously improve the development of nervous system disorders in 0-18 months infants,and can significantly improve these infants' Bayley developmental scale neurobehavioral scores.

Objective To explore the effect of macrolide antibiotics(erythromycin) on tumor necrosis factor(TNF)-α and interleukin(IL)-8 in hyperoxia-induced lung tissue of premature newborn rats,and to study the intervention effect of erythromycin on hyperoxia-induced lung injury.Methods One-day old preterm Sprague Dawley rats were randomly divided into four groups by random number table method:air + sodium chloride group,air + erythromycin group,hyperoxia + sodium chloride group,hyperoxia + erythromycin group.Hyperoxia groups were continuously exposed to oxygen (oxygen ＞ 0.85) and air group in room air.After 1,7,14 days of exposure,the preterm rats of four groups were sacrificed,whole lung of these rats were isolated,the lung histological changes were observed by hematoxylin-eosin staining,TNF-α and IL-8 in pulmonary tissue homogenate were detected by ELISA.Results The results showed that:(1) Compared with air + sodium chloride group,TNF-α and IL-8 expression in hyperoxia + sodium chloride group were significantly increased(P ＜ 0.05) after 1,7 days of exposure [1 d:TNF-α:(16.163 ± 0.574) ng/ml vs.(21.923 ±2.066) ng/ml,IL-8:(18.214 ±3.649) ng/ml vs.(23.546 ± 5.240) ng/ml ;7 d:TNF-α:(15.940 ±0.821) ng/ml vs.(19.688 ±0.764) ng/ml,IL-8:(18.541 ± 4.114) ng/ml vs.(24.255 ±4.692) ng/ml],in particular,TNF-α expression appeared to increase earlier,their expression became significantly weak in 14 days (P ＜ 0.05).(2) Compared with hyperoxia + sodium chloride group,TNF-α and IL-8 expression in hyperoxia +erythromycin group became significantly weak after 1,7,14 days of exposure(P ＜0.05) after the intervention of erythromycin [1 d:TNF-α:(21.923 ± 2.066) ng/ml vs.(18.903 ± 1.851) ng/ml,7 d:IL-8:(24.255 ±4.692) ng/ml vs.(23.508 ±3.543) ng/ml,14 d:TNF-α:(16.443 ±5.466) ng/ml vs.(14.453 ±0.963)ng/ml],but their expression became weaker in 14 days than that in 1,7 days.Conclusion The release of inflammatory mediators TNF-α and IL-8 induced by oxidation outbreak participates in the development of hyperoxia induced lung injury,erythromycin may regulate immune function,inhibits the levels of oxidant-mediated TNF-α and IL-8 induced by oxidation outbreak,and alleviate hyperoxia lung injury in premature rats.

Objective:To explore the efifcacy and safety for radiofrequency catheter ablation (RFCA) of left bundle branch guided by left bundle potential (LBP), X-ray image with EnSiteNavX System in canine model.
Methods:The RFCA of left bundle branch was conducted in 13 dogs. A mapping catheter was positioned in right atrium to record right-sided His-bundle (R-His) potential, and an ablation catheter via right femoral artery was retrograded to left ventriclefor LBP mapping and ablation. Meanwhile, EnSiteNavX System was used to identify R-His, L-His and LBP at the same time. The potential characteristics in dogs with successful ablation were observed, the PR interval, QRS shape and time limit, AH interval, HV interval, the A/V electro-gram ratio in ablationcatheter at before and after ablation were recorded. The procedural time and X-ray exposure time between LBP with X-ray image method and LBP, X-ray image with EnSiteNavX System method were compared.
Results: There were 9/13 dogs received successful left bundle branch ablation, 3 dogs failed and 1 suffered from complete A-V block. At the successful ablation target site, the LBP-V was (17.8 ± 2.6) ms with the range of (13-21) ms, and the A/V electro-gram ratio<1/10. The procedural time and X-ray expose time were signiifcantly decreased in LBP, X-ray image with EnSiteNavX System method than those in LBP with X-ray image method P=0.007 and P<0.001.
Conclusion:Under the LBP, X-ray image with EnSiteNavX System guidance method, left bundle branch could be safely and effectively ablated to establish left bundle branch block (LBBB) model in experimental canine.

Objectives To explore the effect of erythromycin on glutathione hormone (GSH) and γ-glutamyl cysteine synthetase (γ-GCS) in premature newborn rats exposed to hyperoxia, to study the intervention effect of erythromycin on hype-roxia-induced lung injury. Methods One-day old preterm SD rats were randomly divided into four groups:control group, eryth-romycin group, hyperoxia group, erythromycin+hyperoxia group. Hyperoxia group and hyperoxia+erythromycin group were continuously exposed to oxygen (oxygen concentration>0.85), control group and erythromycin group were in room air. Via cau-dal vein, the preterm rats was injected with erythromycin in erythromycin group and hyperoxia+erythromycin group, sodium chloride in control group and hyperoxia group daily. After 1,7,14 day(s) of hyperoxia (or air ) exposure, the preterm SD rats of four groups were killed, whole lung of these rats were isolated and histological changes were observed by hematoxylin-eosin (HE) staining, GSH andγ-GCS of pulmonary tissue homogenate were detected by double antibody sandwich enzyme linked im-munosorbent assay. Total lung RNA was extracted andγ-GCS mRNA was detected by reverse transcription polymerase chain re-action. Results The results showed that:After 1 and 7 day(s) of exposure, the expression of GSH、γ-GCS andγ-GCS mRNA in four groups showed significant differences(P<0.05). Among them, GSH expression in erythromycin + hyperoxia group was higher than that in the other three groups in 1,7,14 day(s) of exposure with significant differences (P<0.05);GSH expression in erythromycin+hyperoxia group and hyperoxia group reached the peak after 7 days of exposure. The expression ofγ-GCS andγ-GCS mRNA in erythromycin+hyperoxia group and hyperoxia group were higher than the other two groups after 1and 7 day(s) of exposure, the expression ofγ-GCS mRNA in erythromycin+hyperoxia group were higher than that of hyperoxia group with significant differences (P<0.05). Conclusions The expressions of GSH andγ-GCS in the lung of premature SD rats were abnor-mal by oxidation outbreak. Erythromycin may increase the activity ofγ-GCS, improve the anti-oxidation ability of GSH, and al-leviate hyperoxia mediated lung injury in premature rats.

Objective To observe the effect of MAP4K4 targeted shRNA on biological characteristics such as proliferation,invasiveness,and apoptosis in human bladder cancer cell.Methods Differentially expressed genes was screened out through cDNA microarray analysis in 5 pairs of fresh-frozen muscle-invasive bladder cancer(MIBC) and adjacent normal tissue obtained from radical cystectomy.Combining the results of genechip and literature review,MAP4K4 was picked up for further analysis.To verify the result of microarray analysis,16 pairs of fresh muscle-invasive bladder cancer (MIBC) and adjacent tissues were assessed for the expression of MAP4K4 mRNA and protein through RT-PCR,qRT-PCR and Western-blot.T24 cell line was stably trasfected with MAP4K4 targeted shRNA and control shRNA,respectively.The effects of MAP4K4 silencing on proliferation,invasiveness and apoptosis of T24 cells transfected with MAP4K4 targeted shRNA and control shRNA were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT),transwell and flowcytometry (FCM) assay.Results MAP4K4 was overexpressed in muscle invasive bladder cancer than in normal tissue.Down regulation of MAP4K4 expression decreased bladder cancer cell proliferation(MAP4K4-targeted versus control,P＜0.001),invasiveness(MAP4K4-targeted versus control,P=0.004)and promoted cell apoptosis(MAP4K4-targeted versus control,P=0.023).Conclusions MAP4K4 is overexpressed in muscle invasive bladder cancer than in normal tissue.Down-regulation of MAP4K4 expression inhibits the invasive ability of bladder cancer.Therefore,MAP4K4 might be a potential therapeutic target for bladder cancer.

Objective To summarize the experience in application of amputation of secondary structures of splenic pedicle and self-made spleen-removing bag in laparoscopic splenectomy the using 3-hole method. Methods The clinical data of 11 patients receiving the procedure from June 2007 to April 2009 in our hospital were retrospectively analyzed. Results Advantages of the procedure were less bleeding, small wound, quick recovery and no occurrence of postoperative complications. Six cases had slight postoperative pain. All 11 patients were cured. Conclusion The technique of amputation of secondary structures of splenic pedicle is safe and feasible in laparoscopic splenectomy using the 3-hole method.

OBJECTIVETo set up a venous congested flap model to study the mechanism of necrosis through long-term microcirculation observation.

METHODSA specially deviced chamber was assembled to one side of the ears in an adult white rabbit, about 7 approximately 10 days after the operation the congested flap model was made and the microcirculatory status of the flap was dynamically observed under a vivo-microscope for a long time.

RESULTSThe venous crisis phenomenon of flap was well studied and the microcirculation of the flap was observed carefully, finally the variational rule of the congestion flap microcirculation was made clear.

CONCLUSIONSThe model could well simulate the venous crisis flap in clinic, and the microcirculation could also be observed for a long time.

Animals ; Disease Models, Animal ; Female ; Male ; Microcirculation ; Rabbits ; Surgical Flaps ; blood supply ; pathology ; Veins ; pathology

OBJECTIVETo investigate the effect of fetal bovine serum (FBS) on expression of pigment epithelium-derived factor (PEDF) in normal epidermal keratinocytes and dermal fibroblasts.

METHODSKeratinocytes and fibroblasts were incubated with 10% FBS. PEDF protein level in the cells was determined by immunofluorescence and Western blot.

RESULTSPEDF was localized mostly in the cytoplasm,while some in the nuclei. The distribution of PEDF in cytoplasm was in a granular pattern. 10% FBS increased the expression of PEDF both in keratinocytes and fibroblasts,but histamine and Phorbol 12-myristate 13-acetate (PMA) did not interfere the distribution of PEDF in cells.

CONCLUSION10% FBS can upregulate expression of PEDF in epidermal keratinocytes and dermal fibroblasts.

Animals ; Cattle ; Cells, Cultured ; Epidermis ; cytology ; metabolism ; Eye Proteins ; genetics ; metabolism ; Fetus ; Fibroblasts ; cytology ; metabolism ; Keratinocytes ; cytology ; metabolism ; Nerve Growth Factors ; genetics ; metabolism ; Serpins ; genetics ; metabolism ; Serum ; physiology ; Skin ; cytology ; metabolism ; Up-Regulation

Gastric cancer is one of the most common malignant tumors. Traditional Chinese medicine (TCM) has been widely used in treatment of gastric cancer, but still lacking large sample controlled trial to evaluate its efficacy.

OBJECTIVETo observe the lethal effect of multidrug resistance gene (MDR1) antisense RNA combined with oxaliplatin and 5-FU on drug-resistant rectal carcinoma cells.

METHODSPC-MDR1 plasmid including MDR1 was constructed with gene cloning techniques. The drug-resistant cancer cells (8348R) were transferred with the plasmids, and the positive neoplasm cells were selected with G418. Green fluorescent protein (GFP) gene was used as a reporting gene to monitor the gene transfer efficiency under the influence of oxaliplatin and 5-FU. The cytotoxicity and therapeutic effects of MDR1 anti-sense RNA combined with oxaliplatin and 5-FU were evaluated by colony-forming rate and MTT assay.

RESULTSA significant decrease of biological activity was observed in 8348R cells transferred with PC-MDR1, cell cycles were blocked in S phase, or in G2/M phase, and apoptosis rate of the cells increased. With treatment of oxaliplatin, the plasmid transfer efficiency in the drug-resistant cancer cells was improved about 18 times. Using an IC(50) dose of oxaliplatin and 5-FU combined with (MDR1) anti-sense RNA, 75 percent of 8348R cells were killed, which was significant higher than that of the control cells.

CONCLUSIONSCombined MDR1 antisense RNA with oxaliplatin and 5-FU has a synergistic effect of killing drug-resistant cancer cells and may be a promising method for treating drug-resistant rectal carcinoma.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Combined Modality Therapy ; Drug Synergism ; Fluorouracil ; pharmacology ; Genes, MDR ; genetics ; Genetic Therapy ; Humans ; Organoplatinum Compounds ; pharmacology ; RNA, Antisense ; genetics ; Rectal Neoplasms ; genetics ; pathology ; therapy ; Transfection