Objective To identify new serum biomarkers of lung adenocarcinoma.Methods Serum samples from 31 patients with lung adenocarcinoma and 31 healthy individuals were applied to SAX-2 protein chips to generate proteomic spectra by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry(SELDI-TOF-MS).The spectra were analyzed with Ciphergen Biosystems software and biomarker patterns software.Results The software identified 102 peaks and m/z 14 022.9 and 3 735.99 were used to construct the classification tree.The classification tree separated adenocarcinoma of lung effectively from healthy individuals,achieving a validity of 100%.The blind test challenged the model with a sensitivity of 100%and a specificity of 100%.Conclusions The results suggest that SELDI-TOF-MS technique can distinguish lung adenocarcinoma patients from healthy individuals and shows great potential for the development of a screening test for the detection of lung cancer.

Background and Objective:Lung adenocarcinoma is,at present,the most common malignancy in the world and its overall 5-year survival rate is disappointing.Because most of the patients are diagnosed at advanced stage,early diagnosis may improve the prognosis of patients with lung adenocarcinoma.Surface-Enhanced Laser Desorption and Ionization Mass Spectrometry(SELDI) is one of the currently used techniques to identify biomarkers for cancers.This study explored the application of serum SELDI proteomic patterns to distinguish lung adenocarcinoma patients from healthy individuals.Methods:Serum samples from 71 lung adenocarcinoma patients,71 healthy volunteers with matched gender,age and history of smoking were analyzed using WCX2 ProteinChip to select potential biomarkers.Proteomic spectra were generated by mass spectrometry.Results:Five highly expressed potential biomarkers were identified with the relative molecular weights of 4047.79?1.60,4203.99?1.91,4959.81?2.13,5329.30?2.55 and 7760.12?4.11.The sensitivity for diagnosing lung adenocarcinoma was 90.14%,78.87%,50.70%,57.75%,73.24%;and specificity was 97.18%,92.96%,70.42%,76.06%,94.37%,when the critical point was made 1.5.Conclusions:SELDI-TOF-MS ProteinChip technology is a quick,easy,convenient,and highoutput analyzing method that it is capable of selecting several relatively specific,potential biomarkers from the serum of lung adenocarcinoma patients and may have clinic value.

Objective To investigate the influences of sodium salicylate on the auditory brainstem response (ABR)and expression of EphA4mRNA in rat inferior colliculus and its effects on salicylate ototoxicity.Methods A total of 30 healthy SPF rats were randomly divided into five groups:the control group (without any treatment),S7 (i.m.injection of sodium salicylate,175mg/kg,twice daily for 7 days),S14(the same method as S7,twice daily for 14 days),S14+R14(the same method as S14,twice daily for 14 days and recovered for 14 days),and S14+R28(the same method as S14,twice daily for 14 days and recovered for 28 days).After the ABR assessment,rats were sacri-ficed after deep anesthesia and the inferior colliculus tissues were dissected.Real-time PCR was used to detect the expression of EphA4mRNA.Results Compared with the control group,ABR thresholds in the S7 group and S14 group were increased significantly (P<0.05),while there was no significant difference between the control group and the S14+R28 group (P>0.05).The inferior colliculus EphA4mRNA expression level of S7 group was signifi-cantly decreased than the control group (P<0.05).The EphA4mRNA expression level of S7 group was significant-ly decreased than the control group (P<0.05),while there was no significantly difference between the control group and the S14+R28 group (P>0.05).Conclusion Long term injection of sodium salicylate can cause changes in the inferior colliculus of EphA4mRNA which are related closely with synaptic plasticity.It may lead the alteration of the inferior colliculus synaptic plasticity,which is associated with the changes of the hearing failure and the tinni-tus behavior.This indicates that EphA4 which is considered as a related protein in the inferior colliculus may play an important role in the pathology of tinnitus.

Objective To investigate the clinical value of multidisciplinary team (MDT) assessment for chronic constipation.Methods The clinical characteristics of 346 patients with chronic constipation who were admitted to the Third Peolep's Hospital of Hangzhou from January 2010 to December 2013 were multidisciplinarily assessed.The muhidisciplinary team was involved surgeons from the department of colorectal surgery,urology,gynecology,psychiatry and psychology,and tools including questionnaires,defecography,anorectal manometry,colon transit study,urodynamic tests,gynecological examination were applied in the study.The measurement data with normal distribution were presented as (x) ± s,the comparison between groups was analyzed using the ANOVA,and the count data were analyzed using the chi-square test.Results Three hundred and forty-six patients who met criteria of this research were selected,including 86 males and 260 females with the ratio of 1 ∶ 3;the mean age was (55 ± 11)years.Of the 346 patients,slow transit constipation accounted for 7.52％ (26/346),defecatory disorder for 60.98％ (211/346),and mixed constipation for 31.50％ (109/346).A total of 93.85％ female patients (244/260) had anterior rectocele,75.43％ (261/346) patients had internal rectal mucosal prolapse,66.76％ (231/346) patients had perineum descending,23.99％ (83/346) had achalasia or inappropriate contraction of internal anal sphincter,18.79％ (65/346) had puborectalis rectocele muscle thickening,5.49％ (19/346) had rectal prolapse.A total of 82.37％ (285/346) patients were involved in other subjects than colorectal surgery.A total of 28.61％ (99/346) patients presented with urinary symptoms,including 65 cases with stress urinary incontinence,23 cases with unstable bladder and 19 cases with bladder neck obstruction (some patients had multiple urological systoms).The incidence of reproductive organ prolapse in female patients was 31.92％ (83/260),the incidence of uterine prolapse and anterior vaginal prolapse were 26.15％ (68/260) and 29.23％ (76/260),respectively.Patients with anxiety and/or depression accounted for 36.13％ (125/346).The male and female patients of slow transit constipation,defecatory constipation and mixed constipation were 10 vs 16,30 vs 79,46 vs 165,respectively,the age was 60 ± 12,56 ± 11,52 ± 10,showing no significant differences (x2=4.046,F =2.877,P ＞ 0.05).In the three kinds of constipation,patients with urinary diseases accounted for 26.92％ (7/26),26.61％ (29/109) and 29.86％ (63/211),patients with gynecological diseases accounted for 11.54％ (3/26),20.18％ (22/109),27.49％ (58/211),patients with psychological diseases accounted for 38.46％ (10/26),39.45％ (43/109),34.12％ (72/211),respectively,showing no significant difference (x2=4.090,P ＞ 0.05).Conclusion MDT assessment for patients with chronic constipation can reflect comprehensively clinical characteristics of chronic constipation,therefore multidisciplinary team should be emphasized in clinical diagnosis and treatment of chronic constipation.

Medicinal plants of the Fructus Amomi containing three species (A momum villosum, A momum longiligu-lare, Amomum villosum var. xanthioides)are well-known, which are widely used as traditional medicines. The mor-phological characteristics of the three origins are very similar, especially in the form of seed. In this study, 60 sam-ples of Fructus Amomi were co llected, and 34 sequences of the Fructus Amomi and their adulterants from GenBank were analyzed. Single nucleotide polymorphisms (SNPs) were detected. All the ITS2 sequences here (including our ex-periments and GenBank data)were examined for SNPs at the interspecies level. Results from the study revealed that two stable bases at position 135 bp and 199 bp were found, which could be used as a unique marker to distinguish the three origins of Fructus Amomi. The two SNPs in the ITS2 were found to exist stably between the three species, and all the GenBank sequences of the Fructus Amomi. Our findings indicated that SNP-based DNA barcoding could be used as an efficient method for the rapid and accurate identification of the three origins of Fructus Amomi.

Objective To investigate the effect of impulse noise expose on the expression of growth associated protein 43(Gap-43) in inferior colliculus in rat.Methods SPF grade Male SD rats were randomly divided into 5 groups.The normal control group received noise exposure.The model groups received an averange impulse noise exposure of 156 dB SPL with a pulse duration of 0.23 ms, once for 6 s, for 50 times.Auditory brainstem responses (ABRs) were measured before and 3,7,14, and 28 d after noise exposure with tone pips of 2,4,8,16, and 32 kHz, from 20 to 110 dB SPL.Bilateral inferior colliculus of rats in the model groups was collected and treated by immunohistochemical staining.Gap-43 expression of rats in different groups was measured by determining the gray value of inferior immunohistochemical images.Results After noise exposure, ABRs threshold in the model groups were significantly higher than those of in the normal group (P<0.05 or P<0.01).ABRs threshold at 14 and 28 days after noise exposure were significantly lower than 3 days after impulse exposure (P<0.05).Expression of Gap-43 in inferior colliculus was significantly up-regulated in the noise exposed groups compared with the normal group (P<0.05 or P<0.01).Expression of Gap-43 was significantly down-regulated 28 days after noise expose compared with 3 days after noise expose(P<0.05).Conclusion Impulse noise exposure leads to significant elevation of ABR thresholds and up-regulation of Gap-43 expression in inferior colliculus.Impulse noise exposure may induce auditory cortex prominent remodeling.

Objective To observe the early effect of organ donation after pancreas-kidney transplantation.Methods Eight cases of diabetic nephropathy received combined pancreas kidney transplantation.There were 8 donors,including 6 males and 2 females,with an average age of (26 10) years old (range from 15 to 42 years).There were 4 cases of donors with China during the transition period of brain heart double death organ donation (C-Ⅲ) standard,3 cases of donors in line with the international standard of brain death organ donation (C-Ⅰ) standard,1 case of international standard of heart death organ donation (C-lⅡ M-Ⅲ) citizen donors.There were 6 men and 2 women for recipients of the same blood type.Results Eight cases were awake 4-6 h postoperation and the ventilator was removed 8-14 h after operation.The rehabilitation therapy began 2 days postoperation from surgery intensive care unit (SICU) to the common wards.Serum C-peptide and insulin levels achieved normal range in 1-2 weeks after transplant.Blood glucose returned to the normal level in 2-3 weeks,and the creatinine level decreased to the normal level in 2 weeks postoperation.Duodenal intramural hematoma occurred in one patient intraoperatively,and the pancreatic graft was removed for safe consideration.Other patients had no serious surgical complications within 2 weeks after transplantation.Conclusion For organ donation after death of pancreas kidney transplantation,early organ function recovered well.Under the strict preoperative evaluation,the young donors can be safely used in combined pancreas and kidney transplantation.

We produced and identified the monoclonal antibodies against Brucella melitensis U‐lipoprotein OMP19 .A DNA fragment coding omp19 of Brucella melitensis was amplified by PCR ,and inserted into the vector of pET‐30a(+ ) ,the result‐ant recombinant plasmid ,which we designated as pET‐30a(+ )/omp19 .We then transformed the plasmid into BL21(DE3) competent cells for the expression of the OMP19 protein .After induction with different concentrations of IPTG ,the colleted cells were analyzed by SDS‐PAGE ,and then OMP19 monoclonal antibodies were prepared through hybridoma technology . These mAbs were tested to reactivity to rOMP19 and nature membrance proteins (NMP) of Brucella melitensis by Western blot and IEST .We successfully constructed an expression vector of pET 30a(+ )/omp19 .An IPTG‐induced expression of the OMP19 protein (19 kDa in molecular weight) was demonstrated by SDS‐PAGE .The fusion protein existed in the form of solu‐ble ,and the OMP19 protein of high purity could be obtained by Ni‐NTA .Western blot assay showed that the refolded protein could be recognized by the anti‐serum against Brucella melitensis .Twenty‐three mAbs to OMP19 was produced in which 91 .30% were IgG1 ,twenty‐two (95 .65% ) mAbs could recognize nature OMP19 protein ,and eighteen (78 .26% ) mAbs could recognize NMP ,four mAbs could react with Brucella melitensis .The protein maintained good immunogenicity and twenty‐three mAbs were obtained ,which we believe provides a good protein candidate for the immunological research .

Objective To establish diagnostic models for pancreatic carcinoma(PC)and to find out the biomarkers related to PC.Methods Serum samples obtained from subjects including PC patients,pancreatic benign disease patients and normal controls were examined with strong anionic exchange chromatography(SAX2)chips for protein profiling using surface enhanced laser desorption/ionization-time of flight-mass spectrometry(SELDI-TOF-MS).The decision tree models and logistic regression models for evaluating the value of serum biomarkers were assessed.SELDI immunoassay and ELISA were used to identify the biomarker and its level in serum respectively.Results Twentysix mass peaks were different between PC patients and normal controls(P＜0.0 1)and 16 mass peaks were different between patients with PC and with pancreatic benign disease(P＜0.05).The decision tree model had a sensitivity of 83.3%and a specificity of 100.0%in differentiation of PC,which was better than that of CA19-9 by ROC curve.There were significant differences in 6 mass peaks among different stages of PC(P＜0.01).Logistic regression model showed a sensitivity of 81.6%and a specificity of 80.6%in diagnosis of early PC.The M/Z 28068 protein was identified as C14orf166 with a sensitivity of more than 82%and a specificity of more than 88%in diagnosis of PC.Conclusions The diagnostic models based on SELDI-TOF-MS were superior to CA19-9 in diagnosis of PC.The identified biomarker C14orf166 is expected to play a role in the diagnosis of PC.

Objective To investigate the factors associated with seizure relapse after antiepilepsy drug (AED) withdrawal in childhood epilepsy.Methods A retrospective analysis was conducted in epileptic children of Hunan Children's Hospital from Jan.2003 to Jan.2011.Among those with anti-epileptic therapy for seizure-free period over 2 years,the patients who relapsed after withdrawal were followed up through outpatient clinic visits and/or by telephone interviews for at least 2 years.Results Of the 127 cases of children enrolled in this study,28 patients(22.05％) relapsed [male:12/59 cases (20.34％) and female:16/68 cases (23.53％)].Cumulative relapse rates were 18.18％ (8/44 cases) in infancy,15.79％ (6/38 cases) in toddlers,23.53％ (8/34 cases) in preschool children,and 54.55％ (6/11 cases)in school age group.Of the patients who relapsed,generalized seizure occurred in 12/87 cases (13.79％),partial seizure in 16/40 cases(40.00％).According to seizure frequency between the first seizure and AED administration,3 cases(6.25％) relapsed among 48 cases of seizure frequency ＜ 5 times,13 cases(24.07％) relapsed among 54 cases of seizure frequency 5 to 10 times,and 12 cases(48.00％) relapsed among 25 cases of seizure frequency more than 10 times.Relapse occurred in 9 cases of monotherapy(9/91 cases,9.89％) and in 19 cases of polytherapy (19/36 cases,52.78％).According to the seizure control period (period between the beginning of antiepileptic treatment and AED withdrawal),14 cases relapsed among 37 cases with the seizure control period of 2 to 3 years (37.84％),8 cases relapsed among 51 cases with the period of 3 to 4 years (15.69％),and 6 cases relapsed among 39 cases with the period of 4 to 5 years(15.38％).According to AED tapering off period,10 cases relapsed among 24 cases with the period of 3 months (41.67％),9 cases relapsed among 36 cases with the period of 3-6 mc ths (25.00％),and 9 cases relapsed among 67 cases with the period of over 6 months(13.43％).Factors associated with an increased risk of relapse were age of epilepsy onset,seizure type,route of administration,timing of antiepileptic trug withdrawal,tapering speed,which were had statistical significance (x =8.051,6.780,16.896,27.607,7.576,8.451,all P ＜0.05).Gender difference was not associated with the risk of relapse(x2 =0.187,P ＞ 0.05).Conclusions Factors associated with an increased risk of relapse are age of epilepsy onset,seizure type,route of administration,timing of antiepileptic drug withdrawal,tapering speed.Standard therapies of early treatment,adherence to medication for at least 3 years,taper period for more than 6 months are associated with a decreased probability for relapse.