Objective · To explore the change of metabolomic profiling after erlotinib (anepithelial growth factor receptor tyrosine kinase inhibitor)resistance of lung adenocarcinoma cells (PC9-ER), and find the differential metabolome associated witherlotinib resistance. Methods · Metabolic profiling of PC9-ER cells and homologous parent PC9 cells was acquired by the ultraperformance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS). The data were analyzed by multi-dimensional statistical methods, such as partial least squares projection to latent structures-discriminant analysis (PLS-DA), to select and identify differential metabolites associated with erlotinib resistance. Results · A total of 14 differential metabolites were identified in PC9-ER cells. Seven up-regulated metabolites included N-acetylspermidine, phosphatidylethanolamine, AMP, pantothenic acid,proline, glutamate, and histidine, while seven down-regulated metabolites included citrulline, phosphorylcholine, glutathione, cysteinylglycine, glutathione oxidized, NAD, and S-adenosylmethionine, mainly participating in glutathione metabolism, glutamate metabolism, ammonia recycling, and protein biosynthesis. Conclusion · Metabolic profiling of erlotinib-resistant lung adenocarcinoma cells was changed. The information of differential metabolites associated with erlotinib resistance could provide clues for new resistance mechanisms and potential metabolism-related drug targets.

BACKGROUND: There are various methods for management of allogeneic bone, xenogeneic bone and various tissue engineered materials, but there is no ideal method for treatment of insufficient bone mass following jaw defects. OBJECTIVE: To observe the repair efficiency of bone composite and biomembrane following large mandibular defect and mandibular defect combined with tooth luxation in animal studies. DESIGN, TIME AND SETTING: The controlled observational animal study was performed at the Animal Laboratory of Zhejiang University from March to July 2006. MATERIALS: The mixed proportion of Bio-oss material and autologous bone powder was 1:1. The proportion of recombinant human bone morphogenetic protein-2 freeze-dry powder dissolved in autologous fresh blood was 0.25 mg:1 mL. Bone powder mixture was moistened by blood containing human bone morphogenetic protein to stick on the medicine spoon for moulding easily. METHODS: Ten New Zealand rabbits were selected. Consecutive bone defects (15 mm×6 mm×5 mm) were made in the inferior border of bilateral mandible body. Bone composite and Bio-gide membrane were randomly implanted into one side (bone composite + Bio-gide membrane group). Another side was directly sutured as blank control group. The remaining 30 rabbits were considered bone composite + Bio-gide membrane + implantation tooth group. A bone defect (15 mm×6 mm×8 mm) was made at the upper site of inferior border of mandible, with the combination of tooth luxation. Bone composite and Bio-gide membrane were implanted, and the luxation teeth were implanted into the original site. MAIN OUTCOME MEASURES: Implantation site, composite conjugation, loose of bone formation and implanted teeth were generally observed. New bone formation at the bone defect site was observed using radiograph and histological method. RESULTS: At 12 weeks following surgery, a bone defect, which was smaller than the original bone, was found at the mandibular defect site in the blank control group. New bones were visible in the mandibular defect site in the bone composite + Bio-gide membrane group. Radiograph demonstrated that the density of defect bone site was similar to normal bone tissue. Histological method revealed that bone implant formed board-shaped bone. No significant loose was detected in implanted teeth of 17 rabbits in the bone composite + Bio-gide membrane + implantation tooth group. Radiograph demonstrated that no transparent area was found in the root tip of 13 rabbits. Histological method showed replacement resorption in 13 rabbits. CONCLUSION: Bone composite combined with Bio-gide membrane for repairing large mandibular defect obtained good efficiency. The outcome of autologous tooth implantation is acceptable in the near future.

Objective To know the epidemiological characteristics of hand-foot-mouth disease in Yangpu District and provide evidence for specific strategies and measures of hand-foot-mouth disease activity prevention and control. Methods Descriptive analysis of the data from hand-foot-mouth disease epidemic situation in Yangpu District from 2005 to 2008 was made. Results 1 348 cases were reported in the whole Yangpu District from 2005 to 2008, and no severe cases, no death. The average morbidity in Yangpu District was 27.48/100 000. The male to female ratio was 1.46∶1. The infection occurred to infants aged from 1 year to 5 years (85.39%). The incidence of the native population children aged 2 to 3 exceeded 10/100 000. The season peak appeared from May to July (70.18%), and outbreaks used to occur at nurseries and kindergartens. The typical clinical presentations mainly included fever and rash. The rash mainly occurred to hands, feet, mouth, buttocks and so on. Conclusions Incidence varied significantly between different sexes, seasons and ages. It can cause large-scale epidemic in a short period of time, the epidemic was very difficult to control, but the leaders attach importance to take the early warning and monitoring, accuratey deal with emergencies, health education promotion and training of comprehensive measures, the epidemic can be effectively controlled.

Objective:To prepare nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR and to examine its cytotoxicity against EGFR positive tumor cells.Methods:By using molecular cloning strategy,prokaryotic expression construct of pET28a-BI7D12-PE38KDEL was generated which consisted of nanobody 7D12 targeting EGFR in the form of a divalent fused with PE38KDEL,a truncated form of pseudomonas exotoxin A via a flexible peptide(G4S)4,and then transformed into E.coli BL21(DE3).Protein expression was induced by adding IPTG,purified by Ni-affinity column chromatography,and verified by Western blot.The binding capacity of the resulted immunotoxin to EGFR-positive cells A549,HT29,MCF-7 and EGFR-negative cells CEM,Jurkat were determined by flow cytometry assay,and its cytotoxicity against the target cells was examined.Briefly,tumor cells were treated with different dosage of the immunotoxin,and the killing efficacy of BI7D12-PE38KDEL on these cells were assessed by WST-1 assay after 72 hours.Results:The SDS-PAGE and Western blot results showed the recombinant immunotoxin BI7D12-PE38KDEL was successfully prepared,and majority of them was expressed in soluble form.BI7D12-PE38KDEL could selectively bind to EGFR-positive cells of A549,HT29,and MCF-7.More importantly,the immunotoxin exhibited much more significant killing effect on these EGFR positive cells compared to the negative control group of CEM and Jurkat cells(P<0.01).Conclusion:In the current study,the nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR was successfully prepared and exhibited a superior inhibition effect for the growth of EGFR-positive cells.

Objective To summarize our experience in hepatic artery reconstruction in adult-to-adult right lobe living donor liver transplantation(LDLT).Methods A retrospective analysis was made for 17 cases undergoing LDLT in our center from May 2007 to Oct 2008.Results All the 17 right lobe graft of the liver was supplied by single right hepatic artery and the mean diameter of right hepatic artery was 3.1 mm.The hepatic artery for segment 4 was mainly originated from left hepatic artery(12/17,70.1%).The recipient right or left hepatic artery was used in 14 cases of reconstruction,proper hepatic artery was used in 2 cases,and gastroduodenal artery was used in one case.Anastomosis was performed with interrupted 8-0 prolene and 12-16 stitches were made on the posterior wall first and then the anterior wall to avoid turning over the vessel.The mean anastomosis time was(51±26) minutes and all hepatic arteries were patent immediately after anastomosis.Hepatic arterial complications including hepatic artery thrombosis (HAT)did not occur after LDLT.Conclusions Detailed evaluation and careful protection of the hepatic artery of segment 4 are the key to successful reconstruction of hepatic artery in LDLT.Anastomosis was performed without flipping the artery wall helped to reduce the difficulty of operation remarkably and with a good result.

Objective To study the effects of melatonin on the expression of endothelin-1 (ET-1) and endothelial nitricoxide synthase (eNOS) in renal cortex of insulin-resistant (IR) rats. Methods Insulin-resistant rat model was established with 6-8 week ages Sprague-Dawwley (SD) rats by high-glucose diet (70% calories from glucose) for 6 weeks. IR rats models were divided into control normal group (CN, n=10), IR group (n=10) and melatonin group (MEL, n=10). Rats in MEL were affused melatonin 10mg/(kg·d) (4PM) for further 6 weeks. The protein and mRNA in rats' renal cortex of each group were assayed by immunohistochemistry and RT-PCR. Results Arterial blood pressure (ABP) and serum triglyceride (TG), free fatty acid (FFA), malondialdehyde (MDA), insulin (Ins) and homeostasis model analysis insulin resistance (HOMA-IR) in MEL were lower than that in IR (all P＜0.01). Serum high-density lipoprotein (HDL-c) and superoxide dismutese (SOD) were higher in MEL than that in IR (all P＜0.01). HE staining showed that the structure of renal cortex in MEL is approximately normal. The levels of ET-1 protein and mRNA in renal cortex of MEL rats were lower than that in IR group. The levels of eNOS protein and mRNA of renal cortex in MEL group were higher than that in IR group (all P＜0.05). Conclusion At the early stage of IR, melatonin application can ameliorate the imbalance between ET-1 and eNOS and protect the angio-endothelial function of renal cortex in high-glucose induced IR rats.

OBJECTIVE:To establish a method for the 5 main components (original syringin,chlorogenic acid,eleutheroside E,isofraxidin and quercetin-3-rhamnoside) in the fruits and roots of wild Acanthopanax senticosus. METHODS:UPLC was per-formed on the column of Waters ACQUITY UPLC HSS T3 with mobile phase of acetonitrile-0.3% phosphoric acid (gradient elu-tion)at a flow rate of 0.2 ml/min. Detection wavelength was 300 nm,column temperature was 30 ℃,and injection volume was 10μl. RESULTS:The linear range was 24.56-184.2 μg/ml for syringin(r=0.9993),18.454-138.405 μg/ml for chlorogenic acid(r=0.9993),8.416-63.12 μg/ml for eleutheroside E (r=0.9997),3.286-24.645 μg/ml for isofraxidin (r=0.9993) and 2.522-18.915μg/ml for quercetin-3-rhamnoside(r=0.9998);RSDs of precision,stability and reproducibility tests were lower than 1%;recover-ies were 99.14%-100.50%(RSD=0.48%,n=6)for syringing in the fruits of A. senticosus、99.03%-100.45%(RSD=0.50%,n=6) for chlorogenic acid in the fruits of A. senticosus、99.22%-100.44%(RSD=0.44%,n=6)for eleutheroside E in the fruits of A. sen-ticosus、99.80%-100.80%(RDS=0.44%,n=6)for isofraxidin in the fruits of A. senticosus、99.76%-101.10%(RSD=0.51%,n=6) for quercetin-3-rhamnoside in the fruits of A. senticosus;99.21%-101.20%(RSD=0.73%,n=6)for syringing in the root of A. senti-cosus、99.81%-101.20%(RSD=0.52%,n=6)for chlorogenic acid in the root of A. senticosus、100.00%-101.50%(RSD=0.62%, n=6)for eleutheroside E in the root of A. senticosus、99.22%-100.40%(RSD=0.47%,n=6)for isofraxidin in the root of A. senti-cosus. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determi-nation of original syringin,chlorogenic acid,eleutheroside E,isofraxidin and quercetin-3-rhamnoside in the fruits and root of wild A. senticosus.

Objective To evaluate the anatomic localization and size of acute necrotic myocardium in the ischemic-reperfused rat hearts using 99m TC-Glucarate and microSPECT/CT.Methods The ischemic-reperfused ( IR) rat heart models were established by ligating left anterior descending coronary artery for 60 min.99mTC-Glucarate was intravenously injected into the rats 24 hours after IR operations .Images were acquired 30 min after administration of 99m TC-Glucarate using microSPECT/CT. Anatomic localization and size of acute necrotic myocardium were analyzed with microSPECT/CT imaging , and these results were compared to those determined by triphenyltetrazolium chloride ( TTC ) staining.Results The microSPECT/CT images showed hot spot accumulations of 99mTC-Glucarate in IR hearts (the heart-to-liver ratio was 1.90 ±0.33), not in controls (P <0.05).The anatomic localization of 99mTC-Glucarate-labeled necrotic myocardium were in correspondence with TTC staining results .The hot spot size was related significantly to necrotic myocardial size determined by TTC staining ( R2 =0.964 ) .Conclusions The localization and size of acute necrotic myocardium can be assessed by non-invasive microSPECT/CT imaging with99m Tc-Glucarate.

Objective:To explore the effect of sarpogrelate on platelet function in patients at the bridging stage before coronary artery bypass grafting (CABG).
Methods: A total of 40 consecutive patients with peripheral artery stent and scheduled for CABG in our hospital from 2011-05 to 2013-04 were enrolled in this study. The patients were randomly divided into 2 groups, Low molecular weight heparin (LMWH) alone group, n=19 and Sarpogrelate+LMWH group, n=21. The medications started at 5-7 days before CABG and stopped at 24 h before CABG. The platelet inhibition rates (platelet aggregation induced by collagen+ serotonin) were examined and compared between 2 groups at the baseline (before randomization), 24h and 1h before CABG respectively.
Results: The platelet inhibition rates were similar between 2 groups at the baseline (87.33 ± 6.82) % vs (86.11 ± 6.87) %, P=0.577 and 1h before CABG (62.60 ± 12.39) % vs (56.19 ± 14.99) %, P=0.148. At 24h before CABG, the platelet inhibition rate in Sarpogrelate+LMWH group was higher than that in LMWH alone group (83.87 ± 8.99)%vs (63.13 ± 10.88)%, P<0.001. Compared with the baseline, the falling range of platelet inhibition was lower in Sarpogrelate+ LMWH group at 24h before CABG, (3.46 ± 6.18) % vs (22.98 ± 9.43) %, P<0.001 and the falling range was similar between 2 groups at 1h before CABG (24.73 ± 14.19)%vs (29.92 ± 14.28)%, P=0.257.
Conclusion: Sarpogrelate + LMWH may result better platelet inhibition rate with quicker recovery of platelet function upon the medication stopping, which might be a feasible management in patients at the bridging stage before CABG.

Objective To investigate the impact of percutaneous coronary interventional (PCI) on the inflammatory indices and postoperative vascular restenosis.Methods This study involved 90 patients undergoing PCI procedures for Coronary artery disease (CAD) compromising a single coronary artery.Fourty healthy individuals with normal findings by coronary angiography were selected as the control group.Before and after PCI or coronary angiography,plasma hs-CRP and IL-6 were measured in all the subjects by immunonephelometry and enzyme-linked immunosorbant assay (ELISA),respectively.Results (1) In the CAD patients,the plasma hs-CRP level was significantly elevated after PCI as compared with the preoperative level((18.69 ±5.14) mg/L vs (14.45 ± 4.32) mg/L,t =1.42,P ＜ 0.01),whereas in the control group,the hs-CRP level underwent no significant changes after coronary angiography((13.59 ±5.99) mg/L vs(12.46 ±5.35) mg/L,t =1.25,P ＞ 0.05).(2) PCI procedures also resulted in significant elevation of plasma IL-6 level in the CAD patients((1.87±0.45) pg/L vs (1.35 ±0.39) pg/L,t =1.33,P＜0.01),but in the control group,IL-6 showed no significant variation after coronary angiography ((1.32 ± 0.41) pg/L vs (1.21 ± 0.38)pg/L,t =1.16,P ＞ 0.05).We observed significant difference of hs-CRP and IL-6 levels between the CAD patient group and the control group (t =4.96,6.61 respectively,P ＜ 0.01).Conclusion Plasma hs-CRP and IL-6 are elevated in CAD patients following PCI procedures.But the roles of elevated hs-CRP and IL-6 in the vascular restenosis following the procedures need further investigation.