Background:Helicobacter pylori(Hp)is an important pathogen for peptic ulcer and gastric cancer,and is reportedly associated with a variety of extragastrointestinal diseases. However,there is no body fluid detection technique for Hp infection in clinical practice. Aims:To identify Hp infection-associated differentially expressed proteins in urine with relative molecular mass more than 10 kDa and provide potential biomarkers for diagnosis of Hp infection through body fluid detection. Methods:Midstream urine was collected from volunteers in the morning,and 13 C-urea breath test was performed to determine Hp infection. Each of 15 Hp-negative and 15 Hp-positive urine samples were mixed respectively for protein extraction. Spectra data were acquired by high performance liquid chromatogram-mass spectrometry,and label-free technology was used for relative quantitative analysis. The other 26 urine samples(15 Hp-negative and 11 Hp-positive) were used for validation by full scan. IPA software was employed for bioinformatics analysis. Results:A total of 475 urinary proteins were detected by label-free quantitative analysis and 42 differentially expressed proteins were identified. Finally,11 significantly up-regulated differentially expressed proteins were confirmed by external scanning validation. Bioinformatics analysis revealed the molecular functions,biological pathways,and related diseases of these differentially expressed proteins. Conclusions:These 11 differentially expressed proteins more than 10 kDa identified in urine might be potential biomarkers for diagnosis of Hp infection and provide molecular evidence for the correlation of Hp infection with extragastrointestinal diseases.

OBJECTIVE:To study the effects of nimesulide combined with oxaliplatin on transplanted tumor growth and im-mune function of rats with esophageal cancer. METHODS:Rats were randomly divided into model group(intragastrically given So-dium carboxymethyl cellulose solution+intravenously given 5% Glucose injection in tail),nimesulide group(intragastrically given 20 mg/kg),oxaliplatin group(intravenously given 13.6 mg/kg in tail)and combination group,10 in each group. Esophageal can-cer Eca109 cells were subcutaneously injected to develop transplanted tumor model. After modeling,rats in each group received rel-evant medicines by corresponding ways,once a day for ig,once every 4 d for iv in tail. Rats were sacrificed after 8 weeks,tumor volume and quality of rats were measured,tumor inhibition rate was calculated,and contents of tumor markers(CEA,CYFRA21-1,SCCAg),percentages of immune cells(CD3+,CD4+,CD8+T cells and NK cell)in peripheral blood were detected. RESULTS:Compared with model group,tumor volume and quality in other 3 groups were decreased (P<0.05);contents of tumor markers were decreased (P<0.05). Percentages of CD3+,CD4+ T cells and NK cell in nimesulide group were increased,percentages of CD8+T cell was decreased(P<0.05). Percentages of CD3+,CD4+T cells and NK cell in oxaliplatin group and combination group were decreased,percentages of CD8+ T cell was increased(P<0.05). Compared with nimesulide group and oxaliplatin group,tu-mor inhibition rate in combination group was increased(P<0.05);contents of tumor markers were decreased(P<0.05);percent-ages of immune cells were lower than nimesulide group and higher than oxaliplatin group(P<0.05). CONCLUSIONS:Nimesulide can enhance the oxaliplatin's antitumor effect on esophageal cancer,and decrease its inhibition degree on immune functions.

Background The current epidemiology study had shown the prevalence of age and sex adjusted dry eye was higher in patients with diabetes than population without diabetes.Further researches demonstrated that the tear film disturbance is common after the phacoemulsification or photocoagulation in the eyes of diabetic patients.Objective The aim of this study was to characterize the clinical features of tear film instability in diabetes patients.Methods One hundred and sixty-two patients with tear-film abnormality referred to Tongren Eye Center from January 1,2010 to September 1,2010 underwent questionnaire about diabetes and other diseases,BUT,Schirmer test.Tear film instability was diagnosed as abnormality of either Schirmer test or BUT and showed as M ( Q25,Q75 ).The right eyes of 162 dry eye patients meeting with the including criteria were enrolled.The patients were assigned to two groups according to with ( 80 patients) or without ( 82 patients) diabetes mellitum.DEQ questionnaire were scored.The percentage of cases with meibomain gland abnormal score ＞ 1 was calculated.Mann-Whitney U analysis and Chisquare analysis were used to compare the difference between the two groups.Results The Schirmer test in diabetic group was 8 ( qualities:5,9 )mm and was longer than 6 ( qualities:5,7 ) mm in non-diabetic patients ( U =2452,P =0.00).The result of BUT test was 3 ( qualities:2,4 ) seconds in diabetic patients and was shorter than 4 (qualities:3,5) seconds in non-diabetic patients( U=2104,P＜0.01 ).The DEQ score of diabetic patients was 15 ( qualities:1 0,19,which was less than21 ( qualities:19,23.25 ) in non-diabetic patients.51.2 ％ ( 41/80 ) diabetic participants and 32.9％ (27/82) nondiabetic participants appeared meibography ( grade larger than 1 ) (x2 =16.07,P=0.00).The percentages of dry eyes were 51.2％ (41/80) and 93.9％ (77/82) respectively in diabetes and nondiabetes groups(x2 =37.24,P＜0.01 ).No significant correlation was found between the diabetes course and DEQ score or meibography( r =0.16,P =0.16 ; r =0.10,P =0.36 ).Conclusions Diabetes patients with tear film instability have longer Schirmer test results,shorter BUT,more severe meibomain glands damage and lower DEQ scores.The dry eye symptom is lack in the diabetic patients though appearing the tear film and meibomain glands damage.Therefore,more attention should be given to ocular surface health in diabetes patients.

OBJECTIVE: To explore the relationship between the expression changes of cytokines, interleukin-6 (IL-6), interleukin-8(IL-8), and tumor necrosis factor-alpha (TNF-alpha), and the wound time following explosive injury to rabbit's chest. METHODS: The rabbit's model of explosive injury was established. The expression levels of IL-6, IL-8 and TNF-alpha in the plasma were detected by ELISA method at different wound time (0.5-12h). RESULTS: The level of IL-6 increased at 3h after wounding and reached peak at 6 h. The level of IL-8 increased at 1 h and reached peak at 6 h. The level of TNF-alpha increased at 0.5 h and reached peak at 3 h. CONCLUSION IL-6, IL-8 and TNF-alpha have a time-related expression after explosive injury.
Animals ; Enzyme-Linked Immunosorbent Assay ; Explosions ; Interleukin-6/metabolism* ; Interleukin-8/metabolism* ; Rabbits ; Thoracic Injuries/metabolism* ; Tumor Necrosis Factor-alpha/metabolism*

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of butoconazole in human plasma. Human plasma samples of 0.2 μL were pretreated by a single step protein precipitation procedure and analyzed using a high performance liquid chromatography (HPLC) electrospray tandem mass spectrometer system. The compounds were eluted isocratically on an Inertsil ODS-SP column (100 mm×2.1 mm, 3 μm), ionized using a positive ion atmospheric pressure electrospray ionization source and analyzed using multiple reaction monitoring (MRM) mode. The ion transitions monitored were m/z 412.8→165.1 for butoconazole and m/z 453.4→230.3 for the internal standard. The chromatographic run time was 3.5 min per injection, with retention time of 2.47 min and 2.15 min for butoconazole and repaglinide, respectively. The method was validated to be linear over the range of 20 to 8000 pg/mL (r>0.999) by using a weighted (1/x(2)) quadratic regression. The mean recovery rate was more than 86.7%, and the intra- and inter-day precision of the quality control samples (QCs) was less than 8.3% and the accuracy ranged from 96.0% to 110.2%, which indicated that the quantitative method was reliable and accurate. The method is simple, rapid, and has been applied successfully to a pharmacokinetics study of butoconazole nitrate suppositories in healthy Chinese females.

Depression is a common emotional disorder that seriously affects people's life and health all over the world. The pathogenesis of depression is complex, and traditional Chinese medicine (TCM) for antidepressants has a good therapeutic effect because of its multi-component, multi-pathway, and multi-target action mode. At present, the anti-depressive mechanism of TCM has not been fully clarified, but it is clear that depression is closely related to metabolic health. Therefore, in order to further explore the anti-depressive mechanism of TCM, this paper proposes research strategies on the anti-depressive mechanism of TCM based on functional metabolomics from the perspective of metabolism, the potential biomarkers of depression are analyzed with the help of multi-omics combined analysis technology, and the functional molecules of TCM for antidepressant are studied. Molecular biology techniques are used to accurately capture the molecular interactions between biomarkers of depression and functional compounds, which identify effective drug targets and further elucidate the biochemical functions and related mechanisms involved in depression metabolic disorders. This paper systematically reviews the research strategies and applications of functional metabolomics in the anti-depressive mechanisms of TCM, expounds on the core value of functional metabolomics, and summarizes the current research status and hot issues of TCM for antidepressants in recent years, providing new methods and new ideas for the study of mechanisms of TCM with the help of functional metabolomics.

Objective To determine the incidence, clinical manifestation and part of lymphokines which represent the balance of Th1 and Th2 in the role of the immunologic mechanisms for IRIS(immune restoration inflammatory syndromes)in patients initiating HAART(Highly Active Antiretroviral Therapy).Methods A prospective study of all patients initiating HAART was performed. A period of six months tracking initiating HAART was performed. The incidence of IRIS, time of occurrence and clinical disease spectrum were recorded. The main T lymphokines including IL-2, INF-γ, IL-4, IL-10 which on behalf of the balance of Th1 and Th2 were detected. To explore the immunopathologic mechanisms for IRIS, the levels of T lymphokines at pre-HAART, initiating HAART for 1 month, 3months and 6 months were compared in IRIS group and non-IRIS group, healthy group. Results A total of 212 patients were enrolled in this study. 59 patients were diagnosed as IRIS at a median of 21 days after HAART initiation (QR 19 days).The main disease spectrum included tuberculosis, herpes virus infections, pneumocystis jirovecii pneumonia. No matter in the IRIS group or non-IRIS group, the main lymphokines baseline of IL-2, INF-γ reduced and IL-4, IL-10 increased before HAART compared to healthy group (P ＜ 0. 05), which had the tendency to restore balance relations initiating HAART. The lymphokines levels had significant difference between baseline and 6 months initiating HAART (P ＜ 0. 05). The changed levels of lymphokines between IRIS group and non-IRIS group before HAART had significant difference compared to healthy group. IL-2, INF-γ increased level[(11.68 ± 2. 89) pg/ml vs (8.52 ±2.26) pg/ml; (22. 19 ± 6. 22) pg/ml vs (18.34 ±5. 35) pg/ml] and IL-10 decreased level [(19. 21 ± 4. 03) pg/ml vs (23. 19 ± 5.92) pg/ml] had significant difference between IRIS group and non-IRIS group initiating HAART I month(P ＜0. 05). Conclusions The incidence of IRIS during 6 months initiating HAART in HIV/AIDS was 27. 8%, IRIS usually occurred in 1 month initiating HAART. The most common disease spectrum was infectious disease, including tuberculosis and herpes virus infection. Lymphokine of Th1 and Th2 existed unbalance in IRIS group and non-IRIS group before HAART. The unbalance tendency in IRIS group was more obvious. All lymphokines had the trend to recover balance. IL-2, INF-γ significantly increased and IL-10 significantly decreased, which might involve the occurrence of the IRIS.

Objective To research the differential expression of trace phosphorylated proteins in human gastric adenocarcinoma epithelial (AGS) cells infected by Helicobacter pylori. Methods H. pylori 26695 strain infected AGS cells 4 h and AGS cells was cultivated for 4 h as a comparison. The proteins of AGS and comparison AGS cells were extracted. Their phosphorylated proteins were enriched by metal ion af-finity adsorption enrichment techniques. After desalinated and purified the phosphorylated proteins samples were separated by 2-dimensional polyacrylamide gel electrophoresis (2-DE) technique. Computer assisted image analysis was used to analyze the differential proteomic expression. The significantly differentially ex-pressed proteins were unambiguously assigned identities by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/TOF). Results Fifteen kinds of proteins were down-regulated, 4 kinds of new proteins were observed, 1 kind of proteins were up-regulated, 1 kind of proteins unexpression. The 21 proteins that were significantly differentially expressed , including cellular calcium ion homeostasis, transcription, interpretation, protein folding and transport, ribosomal assembly, centrosome replication, chromosome stability, cellular structure, cellular proliferation and apoptosis. Conclusion H. priori can cause a wide range change to human gastric adenocarcinoma epithelial cell protein pheshorylation. This change character has great significance to further comprehensive understanding of the pathogenesis of H. pylori.

Objectives To study the performance of different microbial automated inoculation systems and to evaluate the performance of the Probact microbial automated inoculation and incubation system ( Probact system) and its applications in clinical microbiology laboratory.Methods A total of 160 clinical specimens, including respiratory secretions ( n=61 ) , urine ( n=49 ) , and feces ( n=50 ) , that were submitted to the Clinical Microbiology Laboratory in Peking Union Medical College Hospital of Chinese Academy of Medical Sciences from February 2015 to April 2015 were evaluated.These specimens were processed with conventional manual method, the Probact automated inoculation system, and PREVI Isola Inoculator.The quantity of bacterial species recovery, number of effectively isolated colonies, total number of colonies recovery per plate, and time of processing the 160 specimens by the three methods were evaluated. Wilcoxon signed-rank test and Kruskal-Wallis rank sum test were used for statistical analysis.Results The Probact system had significantly higher quantity of bacterial species recovery (respiratory specimens 3.41 ±1.40, urine 1.92 ±0.86, and feces 1.16 ±0.79) than those by the Isola Inoculator (respiratory specimens 3.75 ±1.29, urine 2.24 ±0.97, and feces 1.92 ±0.72), (P=0.006, 0.011, <0.001).Compared to the manual method, Probact performed less quantity of bacterial species recovery for respiratory specimens(3.85 ±1.38), but higher in feces(0.80 ±0.81)( P<0.001).There is no significant differences for urine ( 1.84 ±1.23 ) ( P=0.266 ) .As for number of isolated colony, the Probact system ( respiratory specimens 12.16 ±7.72, urine 2.71 ±4.24, and feces 5.40 ±5.04 ) had significant smaller numbers than that of Isola Inoculator (respiratory specimens 16.56 ±5.76, urine 4.35 ± 4.89, and feces 8.40 ±3.70) (P<0.001,0.007,0.003).However, both system had larger numbers of isolated colonies than those by the manual method (respiratory specimens 11.30 ±8.42, urine 2.67 ±4.34, and feces 1.90 ±3.90) and the difference was significant for fecal specimens(P<0.001).Regarding the total number of colonies recovery, larger number was found by Isola Inoculator than that by the Probact system for fecal specimens, however, there were no significant differences for respiratory or urine specimens (P=0.524,0.738).Compared with manual method, the Probact system had significantly more numbers of colonies recovery for respiratory and fecal specimens ( P<0.001 ) . The total time for processing 160 specimens was shortest for manual method (281 min), followed by Probact system (419 min) and Isola Inoculator (495 min) .Conclusions The performance of the Probact system is better than the manual method but no superior to the Isola Inoculator.The Probact system can meet the clinical need in terms of full automation and standardization of specimen inoculation and prevention of bias of processing by laboratory staffs using manual method.

Objective To investigate the prognostic factors of cervical high-grade squamous intraepithelial lesions treated by cold knife conization with negative margin.Methods Two hundred and sixty-six women with cervical high-grade squamous intraepithelial lesions treated by cold-knife conization with negative margins at Beijing Hospital between Jan 1999 and Jan 2004 were analyzed retrospectively.All patients were followed up with cytology,high-risk human papillomavirus(HPV)test and eolposcopy if necessary.Results The cervical CIN recurrence rate was 8.6% with no incidence of invasive cervical cancer after a median follow-up of 46 months.The recurrence was related to the grade of lesions and gland involvement pathologically.One of 20(5.0%)cases with cervical intraepithelial neoplasia(CIN)Ⅱ,9 of 164(5.5%)cases with CIN Ⅲ(excluding carcinoma in situ,CIS)and 13 of 82(15.8%)cases with CIS recurred(P