Objective To determine the expression of serum proteins in pancreatic cancer patients based on mass spectrometry to screen differential proteins and to find potential molecular biomarkers to prediagnosis of pancreatic cancer. Methods The technique of isobaric tags for relative and absolute quantitation (iTRAQ) and two-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS) were used to analyze the serum proteins in 15 pancreatic cancer patients and 10 healthy controls. Screening for serum differential proteins was performed via retrieving Panther, Mascot and Scaffold databases. Ingenuity Pathway Analysis was used to detect the correlation between proteins. Results The expression level of 442 proteins was quantified, and 76 proteins were found to be differentially expressed which changed corresponding biological process. The up-regulation of DNA repair protein 50 (RAD50) and down-regulation of transforming growth factor β1 (TGFβ1) and apoptosis protease-activating factor 1 (APAF1) which were correlated with cell growth and apoptosis were found in pancreatic cancer. A closely interacted network was formed between these three differential proteins and other molecules, which could effect metabolism in pancreatic cancer patients. Conclusion Differential expression of serum proteins screened by iTRAQ in pancreatic cancer patients may be the potential markers of pancreatic cancer, which can provide molecular basis for early diagnosis of pancreatic cancer.

10.3969/j.issn.2095-4344.2013.23.018

Objective To investigate the anaesthetic effect of propofol combined with remifentanil by target-controlled infusion (TCI) used in gynecological laparoscopic myomectomy.Methods Fifty cases,who were scheduled for gynecological laparoscopic myomectomy in our hospital from June 2010 to June 2011,was randomly divided into propofol combined with remifentanil group (n =25) and inhalation anesthesia (remifentanil combined with sevoflurane) group (n =25).The heart rate and blood pressure were recorded at the time of before induction of anesthesia (T0),30 min after carbon dioxide pneumoperitoneum,the end of operation and 3 min after extubation.The awake time,time of extubation and surgery time were also recorded.Results The hemodynamics were kept stable in propofol combined with remifentanil group,and there were no significant difference with respect to SABP,DABP and heart rate at all time points compared with baseline (P ＞0.05) in propofol group.However,in inhalation anesthesia group,SABP,DABP and heart rate were increased significantly at 30 min after carbon dioxide pneumoperitoneum and 3 min after extubation when compared with baseline (P ＜ 0.05) and were higher than those of propofol group (P ＜ 0.05) at counterpart time points.In inhalation anesthesia group,the awake time ((9.3 ± 1.5) min vs (4.9 ± 1.1) min,t =10.56,P =0.017) and time of extubation ((12.9 ± 2.4) min vs.(6.8 ± 1.2) min,t =12.36,P =0.01) were significantlv longer than that of propofol group (P ＜ 0.05).Conclusion Propofol combined with remifentanil TCI-based anesthesia could achieve the optimal hemdynamic stability during anesthesia maintance and better recovery profile from anesthesia in gynecological laparoscopic myomectomy.

Background The special pathological change of diabetic retinopathy(DR)is microvascular disorder.Angiopoietin-like protein 2(Ang-2)is a new protein associated with genesis of blood vessels.Quercetin has multiple pharmacological action,including improving the microcircularion and the permeability of blood capillary.However,the action mechanism of Ang-2 on DR was unclear.Objective The present study was to investigate the effects of quercetin on Ang-2 and its receptor Tie2 expression in retina with diabetes mellitus.Methods Sixty clean male Wistar rats were randomized into 7 groups and 10 rats for each group,and 10 rats served as blank control group.Streptozotocin of 35 mg/kg was intraperitoneally injected in 60 rats to establish the diabetic models.Quercetins encapsulated by liposome with the doses of 50,150 and 250 mg/(kg · d)(3-5 ml)were used to gavage in different groups of models for 12 weeks,and normal saline solution and calcium dobesilate were used at the same fashion as the negative control group and positive control group,respectively.Twelve weeks later,the animals were sacrificed and retinas were isolated.Expressions of Ang-2 protein and Tie2 mRNA in retinas were detected by ELISA and RT-PCR,respectively.The usage and rearing of the animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Sciences and Technology Commission.Results ELISA showed that the A450 of Ang-2 in 150 and 250 mg/(kg · d)quercetin groups was 0.796±0.057 and 0.842±0.043 respectively and was lower than that negative group(1.012±0.046),showing statistically significant differences(q =2.95,2.698,P＜0.05).RT-PCR assay showed that expression of Tie2 mRNA(Tie2 mRNA/GAPDH mRNA)in retinas was 0.712±0.092 and 0.821±0.087,presenting statistically significant differences in comparison with negative group(1.182±0.098)(q =3.497,2.852,P＜0.05).The expression levels of Ang-2 and Tie2 mRNA in retina were lowest in 150 mg/(kg · d)quercetin group.Conclusions Quercetin can improve the retinal microcirculation by downregulating the expressions of Ang-2 and its receptor in early period of diabetic rats.

Objective:To explore the curative effect and safety of febuxosta for the patients with primary gout complicated with mild to moderate renal insufficiency.Methods:60 male patients with primary gout complicated with mild to moderate renal insufficiency were enrolled in the affiliated Hospital of integrated traditional Chinese and Westem Medicine,Nanjing University of Chinese Medicine from January 2015 to August 2016 and randomly divided into two groups,the control group (n=30) was treated by allopurinol,and the study group (n=30) was treated by febuxosta treatment.The blood uric acid (BUA) and hepatorenal function were compared in the treatment period,and the incidence of adverse reactions and the recurrence of gout in post-treatment were compared.Results:At 1,2 and 3 month after treatment,the serum level of BUN in the study group was significantly lower than that of the control group (P＜0.05).The renal function of all patients were significantly improved compared with pre-treatment (P＜0.05),but the no significant difference was found between two groups at the same time(P＞0.05).The control group showed the adverse reactions with limb discomfort,drowsiness,nausea,abdominal distension,diarrhea and itch of skin,and the study group showed limb discomfort,drowsiness and abdominal distension,and one patients presenting allergic skin rash and discontinuation of allopurinol.The total incidence of adverse reactions in the study group was significantly lower than that of the control group (P＜0.05).Conclusions:Febuxosta could effectively and safely decrease the blood uric acid levels and improve the renal function of patients with primary gout complicated with mild to moderate renal insufficiency.

Objective To evaluate the effects of recombinant human neuregulin-1? on a rat model of adriamycin-induced dilated cardiomyopathy(ADR-DCM).MethodsMatched adult male Sprague-Dawley rats were administered with 2.5 mg/kg adriamycin intravenously once a week for 10 weeks to establish models of ADR-DCM.The ADR-DCM rats were randomly divided into 3 groups as follows: the rhNRG-1? group(rhNRG-1?,20 ?g/kg,i.v.,once a day for 10 d,n=8),the negative control group(excipient,n=8),and the positive control group(digoxin,0.062 5 mg/kg,i.v.,once a day for 10 d,n=7).Another 8 normal SD rats in the same situation served as normal control group(control,n=8).Their heart function and hemodynamics were evaluate after the treatment.The ratio of heart weight/body weight was calculated and the thickness of LV parietes was measured.The pathological lesions of cardiac muscle tissues were evaluated after HE staining.Brain contents of natriuretic peptide(BNP) and troponin T(TnT) were determined.The animal treatment was all accorded to the ethical standards.ResultsCompare with the normal control group,the LVESD and LVEDD of ADR-DCM rats were increased(P

Porcine circovirus 3 (PCV3) has been detected in major pig-producing countries around the world since its first report in the US in 2016. Most current studies have focused on epidemiological investigations and detection methods of PCV3 because of lack of live virus strains for research on its pathogenesis in porcine cells or even in pigs. We constructed a recombinant plasmid pCMV-Cap carrying the PCV3 orf2 gene to investigate the effects of capsid (Cap) protein expression on autophagic response in human embryonic kidney cell line 293T (HEK293T). We demonstrate that PCV3 Cap protein induced complete autophagy shown as formation of autophagosomes and autophagosome-like vesicles as well as LC3-II conversion from LC3-I via inhibiting phosphorylation of the mammalian target of rapamycin (mTOR) in HEK293T cells. The ubiquitin-proteasome pathway is also involved in the autophagy process. These findings provide insight for further exploration of PCV3 pathogenetic mechanisms in porcine cells.

OBJECTIVETo observe the vasodilating effect of adrenomedullin 2 (ADM2) by antagonizing angiotensin 1 (Ang II), and to explore its mechanism.

METHODSEighteen male, 180-200 g SD rats were randomly divided into 3 groups (n = 6): control group, Ang II (150 ng/(kg x min)) group and Ang II (150 ng/(kg x min)) + ADM2(500 ng/(kg x h)) group. Mini-osmotic pumps filled with peptide were implanted in the back of rats subcutaneously. After two weeks, the blood pressure was measured by the way of carotid intubation. The plasma was collected for the detection of nitric oxide (NO) content and the activity of endothelial nitric oxide synthase (eNOS). The in situ oxidation of fluorescent dye dihydroethidium (DHE) was used for detecting superoxide in rat arteries. The rat isolated arterial rings were made for studying the vasodilating effect of ADM2. Human umbilical vein endothelial cell line EA. hy 926 cells were cultured and their intracellular reactive oxygen species (ROS) were evaluated by probe DCFH-DA.

RESULTSADM2 dramatically decreased the blood pressure in angiotensin II-induced hypertension rat model, enhanced plasma NO content and the activity of eNOS and reduced superoxide formation in vessel walls. ADM2 also induced relaxation of the vascular rings preconstricted by Ang II in a concentration-dependent and endothelium-dependent manner. In cultured vascular endothelium, ADM2 ameliorated the ROS generation induced by Ang II.

CONCLUSIONAdrenomedullin 2 relaxed blood vessels by antagonizing angiotensin II-induced oxidative stress and improving the vascular endothelial function.

Adrenomedullin ; pharmacology ; Angiotensin II ; pharmacology ; Animals ; Antihypertensive Agents ; pharmacology ; Blood Pressure ; drug effects ; Drug Antagonism ; Endothelium, Vascular ; drug effects ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Male ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type III ; blood ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Vasodilation ; drug effects

In order to set up a base for stem cells to be widely used in clinical medicine, we tried to optimize, in this study, the technique that induces human mesenchymal stem cells (hMSCs) to differentiate into neural stem cells by using cerebrospinal fluid (CSF) from the different groups. After the induction, presence of neural stem cells was confirmed with microscope observation, flow cytometry analysis, immunohistochemistry and fluorescent immunohistochemistry. At the same time, we also compared and analysed the data of the number of stem cells when it totally met the requirements for clinical treatment and the days required. At last, we confirmed that hMSCs could be induced to differentiate into neural stem cells, and that the number of cells totally met the requirements for clinical treatment. But there were some differences both in the number of cells and the days required. Among the groups, the group that marrow mesenchymal stem cells from patients own induced by CSF from healthy volunteers used the shortest time and the quantity of the cells was significantly higher than those of the others.
Cell Differentiation ; Cerebrospinal Fluid ; chemistry ; Culture Media ; chemistry ; Flow Cytometry ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; cytology ; Neural Stem Cells ; cytology

Objective To use the matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) technique for detecting the mutation gene of neonatal non-syndromic hereditary hearing impairment gene in Guangxi and to investigate its effectiveness and feasibility in clinical application.Methods A total of 7 100 newborns were performed the hearing preliminary screening and secondary screening by adopting AABR.The genomic DNA was extracted by the heel blood spot.Twenty mutation characteristics of 4 deaf predisposing genes were detected by MALDI-TOF-MS.Results The pass rate of hearing screening in 7 100 newborns was 97.11％ (6 895/7 100),the positive rate of neonatal gene mutation was 3.54％ (251/7 100),in which the GJB2 gene mutation was in 131 cases,the carrying rate was 1.84％,235delC heterozygous mutation was in 108 cases.SLC26A4 gene mutation was in 93 cases,which dominated by 1229C＞T heterozygous mutation and IVS7-2A＞G heterozygous mutation,mtDNA12SRNA gene mutation was in 16 cases and GJB3 gene mutation was in 11 cases.Conclusion Adopting the MALDI-TOF-MS screening technique can increase the detection rate of hot point mutation in common deaf related genes and discover neonatal genetic NSHI from molecular level and provides the corresponding geneticconsulting guidance for early finding and predicting deaf occurrence,and formulating the interventional measures.