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.

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.

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.

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

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 study the expressions of Cx43,CD105 and VEGF in HBV related HCC tissues and the relationships between Cx43 expression and recurrence and prognosis after cancer radical resection in HCC patients stratified by serum AFP levels. Methods The expressions of Cx43,CD105,VEGF in 234 HBV related HCC tissues were examined by tissue microarray and two-step methods of PV-6000 of immunohistochemistry and the expressions of Cx43 in 20 frozen HCC specimens were examined by RT-PCR. Results Cx43 in HCC tissues was positive as examined by both immunohistochemistry and RTPCR methods.Positive Cx43 expression is correlated with lower early recurrence ( Log Rank P =0.001 ),longer disease free survival (Log Rank P =0.026 ) and overall survival( Log Rank P =0.000 ) as showed by the Kaplan-Meier analysis in patients with AFP ＜ 400 μg/L. The expression of Cx43 is an independent prognostic factor.The positive expression of Cx43 related with lower positive expression of CD105 and VEGF (P =0.018,0.023 ),and correlated with histological differentiation (P =0.002),the number of focus (P =0.033 ),blood vessel tumor embolism ( P =0.029 ). Conclusions The expression of Cx43 is correlative with the expression of CD105 and VEGF,and is predictive of HCC early recurrence and poor prognosis after radical hepatectomy in HBV related HCC patients with serum AFP ＜ 400 μg/L.

This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.

Objective:To observe the clinicopathological features of gastrointestinal stromal tumor (GIST) cases with concurrent carcinoma. Methods:Patient data of 24 GIST cases with concurrent carcinoma were collected from the 157 GIST cases reported be-tween 2002 and 2012. The clinicopathological features of the GIST cases with concomitant carcinoma were studied. The expression of CD117, CD34, and SMA by the tumors was assayed using the immunohistochemical EliVision method. In particular, the expression of the proliferation marker Ki-67 was studied. Results:GIST cases with concurrent carcinoma accounted for 15.3%of the total GIST cas-es studied. The GIST patients with concurrent carcinoma included 14 males and 10 females. The male-female ratio of these patients was 1.4∶1. The age of the patients ranged from 41 years to 66 years, with a median age of 55 years. Lesions at the inferior segment of the esophagus were found in 7 of the 24 selected GIST cases;lesions at the gastric wall and in the intestines were observed in 15 and 2 cas-es, respectively. The diameter of the GIST cases with concurrent carcinoma ranged between 0.6 and 3.8 cm, with an average of 1.50 ± 0.85 cm. Slight dysplasia was observed in 4 of the 24 cases; no heteromorphism was present in the remaining 20 cases. The mitotic counts of GIST cases with concurrent carcinoma ranged from 0/50 HPF to 5/50 HPF, with an average of (0.79±1.83)/50 HPF. The pro-liferative index of Ki-67 in the GIST cases with concurrent carcinoma ranged between 0 and 7.72, with an average of 2.16 ± 3.26. The concurrent carcinoma cases included 5 cases with esophageal carcinoma, 2 with cardiac carcinoma, 15 with gastric cancer, and 2 with intestinal cancer. In contrast to the GIST cases with concurrent carcinoma, the GIST cases without carcinoma complications included 74 males and 59 females. The male-female ratio was 1.25∶1. The age of the patients without concurrent carcinoma ranged from 43 years to 71 years, with a median age of 54 years. Among the 133 GIST cases without cancer complications, gastric, intestinal, and esophageal lesions were found in 114, 13, and 6 cases, respectively. The diameter of GISTs without cancerous complications ranged from 2.4 cm to 15.5 cm, with an average of 6.11 ± 7.09 cm. Different degrees of dysplasia were seen in 82 of the 133 cases. The mitotic counts in the GIST cases without cancer complications ranged from 0/50 HPF to 53/50 HPF, with an average of (3.81±23.67)/50 HPF. The prolifera-tive index of Ki-67 for these cases ranged from 0 to 39.21 and averaged at 6.22 ± 16.96. The male-female ratio of the GIST cases with cancer complications was higher compared with the GIST cases without. The average diameter of GISTs with complications was small-er compared with that of GISTs without complications. The mitotic counts and the proliferative index of Ki-67 were significantly lower in the GIST cases with cancer complications than in those without (t=1.981, P<0.05 vs. t=1.993 5, P<0.05). Conclusion:Concurrent car-cinomas were found in 15.3% of the total GIST cases. No special clinical symptoms were observed in most GIST cases with cancer complications, as revealed when the carcinomas were examined. The proliferative index of Ki-67 in the GIST cases with concurrent car-cinoma is significantly lower compared with that of the GIST cases without complications.