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 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 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 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.

In order to verify the sstability and accuracy of DNA barcode technique, we chose Rosa laevigata Michx as study object. Genomic DNAs of 10 samples were extracted by modified CTAB method. ITS2 sequences were obtained by direct PCR sequencing; the other 6 sequences were obtained from GenBank. The sequences were assembled using the CodonCode Aligner. All of the 16 ITS2 sequences were aligned through Clustal-W and the genetic distances were computed using MEGA 5.0 in accordance with the Kimura 2-parameter (K-2-P) model. Results indicated that the lengths of ITS2 regions of R. laevigata ranged from 219 to 221 bp with two Poly C structure in it. The intra-specific genetic distances were smaller than inter-specific ones in ITS2 regions of R. laevigata. The NJ tree showed that R. laevigata and adulterants were divided into two clades, with 99%bootstrap value, showing good monophyly. So, ITS2 was considered a good marker to identify R. laevigata and its adulterants.

Objective To determine the prevalent herpes simplex virus (HSV) strain in patients with genital herpes (GH),and to evaluate the sensitivity,specificity,positive predictive value (PPV) and negative predictive value (NPV) of anti-herpes simplex virus type 2 (HSV2) IgG and IgM antibodies in the diagnosis of genital herpes (GH) before in vitro fertilization (IVF).Methods Totally,193 HSV2 clinical strains isolated in cell culture from the lesions of patients with GH in the Department of Dermatology,First Affiliated Hospital,Sun Yat-sen University between 2009 and 2011 were typed by using type-specific fluorescein isothiocyanate (FITC)-labelled anti-HSV monoclonal antibodies.Serum samples were obtained from 57 anti-HSV2 IgM/IgG antibody-positive females with suspected GH as well as their husbands (clinical observation group),68 HSV culture-positive patients diagnosed with GH (positive control group),and 120 children aged 8-12 years (negative control group).Enzyme-linked immunosorbent assay (ELISA) was performed to detect anti-HSV1/HSV2 IgG/IgM antibodies in these serum samples.Statistical analysis was carried out using chi-square test.Results There was a significant difference between the positive control group and negative control group in the positivity rate of anti-HSV1 IgG (89.71％ (61/68) vs.40.80％ (49/120),P ＜ 0.01) and anti-HSV2 IgG (91.18％ (62/68) vs.0,P ＜ 0.01),but not in that of anti-HSV1 IgM (20.59％ (14/68) vs.21.70％ (26/120),P ＞ 0.05) or anti-HSV2 lgM (13.24％ (9/68)vs.13.30％ (16/120),P ＞ 0.05).In the clinical observation group,the positivity rate of anti-HSV1 and anti-HSV2 IgM antibodies,anti-HSV1 and anti-HSV2 IgG antibodies was 80.70％ (46/57),91.23％ (52/57),84.21％ (48/57) and 14.04％ (8/57) respectively in the females,19.30％ (11/57),8.77％ (5/57),87.71％ (50/57),12.28％ (7/57)respectively in the males,with significant differences in the positivity rate of anti-HSV1 and-HSV2 IgM antibodies (both P ＜ 0.01),but not in that of anti-HSV 1 or-HSV2 IgG antibodies (both P ＞ 0.05).The sensitivity,specificity,PPV and NPV were 13.24％ (9/68),86.67％ (104/120),36.00％ (9/25) and 63.80％ (104/163) respectively for anti-HSV2 IgM antibody in the diagnosis of GH,91.18％ (62/68),100.00％ (120/120),100.00％ (62/62),and 95.24％ (120/126) respectively for anti-HSV2 IgG antibody.Conclusions HSV2 prevails in the patients with GH in this region,while HSV1 only amounts to 5.18％.The type-specific anti-HSV2 IgG antibody shows a higher specificity,sensitivity,PPV and NPV in the diagnosis of GH than anti-HSV2 IgM antibody,hence,the type-specific anti-HSV2 IgG antibody is superior to anti-HSV2 IgM antibody in diagnosing GH before assisted reproduction.

Objective To detect the disparity of three biological molecules Caveolin-1,IL-1β,VEGF in cerebrospinal fluid of children with viral encephalitis at the different stages; to explore the role of Caveolin-1,IL-1β,VEGF in the pathogenesis of viral encephalitis;and to evaluate their clinical significance in assessing the severity and prognosis of viral encephalitis.Methods We recruited 65 inpatients children with viral encephalitis in the Second Neurology Department of Hunan Children's Hospital from July 2011 to July 2012.Subjects were divided into 2 groups:54 cases of acute phase and 11 cases of recovery phase.According to the clinical manifestations,they were re-divided into 40 patients with mild viral encephalitis and 25 cases of severe viral encephalitis.Twenty healthy age matched controls (10 cases of epilepsy and 10 cases of congenital abnormality) were also taken for the study.Cerebrospinal fluid exam,EEG,head MRI and other tests were performed in all patients.Caveolin-1,IL-1β and VEGF levels in cerebrospinal fluid of 65 children with viral encephalitis and 20 age-matched controls were measured using ELISA.Results Cerebrospinal fluid Caveolin-1,IL-1β,VEGF levels in the acute phase of viral encephalitis were (49.209 ± 22.320) pg/ml,(16.923 ± 6.823) ng/ml,(44.342 ± 19.264) ng/ml respectively,and (33.253 ± 20.349)pg/ml,(11.724 ± 3.009)ng/ml,(30.312 ± 18.147) ng/ml in recovery phase,which were significantly higher than those of controls (P ＜0.01).The difference was statistically significant between acute phase and recovery phase (P ＜ 0.05).Acute viral encephalitis patients had higher Caveolin-l,IL-1β,VEGF levels than the epilepsy group,and the difference was statistically significant (P ＜ 0.05).In viral encephalitis group,children with cerebrospinal fluid protein content (0.5 ～ 1.0 g / L) had higher of Caveolin-1,IL-1β and VEGF levels as compared with those with cerebrospinal fluid protein content ≤ 0.5 g/L,and the difference was statistically significant (P ＜ 0.01).Cerebrospinal fluid Caveolin-1,IL-1 β and VEGF showed no significant difference among children with different severity of encephalitis,different levels of frequent seizures,different degrees EEG changes (P ＞ 0.05).But in the patients with severe head MRI changes,cerebrospinal fluid Caveolin-1,IL-1β,VEGF levels increased significantly (P ＜ 0.05).Conclusions Caveolin-1,IL-1β and VEGF may participate in the pathogenesis of viral encephalitis.Detection of these parameters may be helpful to the evaluation of the severity and prognosis of viral encephalitis.

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 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.