Objective To explore the objectivity and scientificity of fecal sampling , and to provide reference for investigating the relationship between intestinal microbes and diseases . Methods Terminal restriction fragment length polymorphism, degeneration gradient gel electrophoresis and real time fluorescent quantitative PCR techniques were applied to differentially analyze the bacterial community composition and abundance of intestinal contents and feces taken from dif -ferent sites of BALB/c mouse intestine .Results The predominant T-RFLP fragments ( T-RFs) in feces in the rectum and colon were 244 bp, 255 bp and 449 bp, however , those in feces of the small intestine including duodenum , jejunum and il-eum were 60 bp, 73 bp, 261 bp, 268 bp and 272 bp, and with a larger variation of the bacterial community composition in various parts of the small intestine .The bacterial abundance in the contents of duodenum and jejunum were 6.9 log ( cop-ies)/g and 8.3 log (copies)/g, fewer than in the other parts of the intestine , while the bacterial abundance in the feces was as high as 11.8 log (copies)/g, being about 2 times higher than that in the duodenum and jejunum (P<0.05), and similar to that in the ileum and colon content (P>0.05).Conclusions The inter-mouse variations of bacterial communi-ty composition in the large intestine contents are small .The bacterial composition and abundance are similar suggest that studies on the relationship between large intestine especially colorectal microbiota and diseases may be conducted via fecal sampling.

Objective To investigate the antitumor effects of IL-18 gene transfected T cells on pancreatic cancer cell SW-1990.Methods Construction of IL-18 contained recombinant lentivirus using PCR method and packaged at HEK293T cell, then transfected the human T cells and evaluated the antitumor effects when cocultured with SW-1990.Results The IL-18 gene contained recombinant lentivirus was successfully constructed and packaged, and transfected the T cells, the LDH secretion and IL-2 and IFN-γ content all increased significantly (P<0.01) when cocultured with SW-1990 cell.Conclusions T cells transfected with IL-18 possessed more potent antitumor effects to pancreatic cancer cell SW-1990 as compared to the regular T cells.

ObjectiveTo explore the inductive action of docosapentenoic acid(DPA) on neurite outgrowth in PC12 cells in vitro.MethodsNeurite outgrowth in PC12 cells was examined after the treatment with different concentration of DPA using Motic Zamges Plus software mapping cell image system.Western blot was performed to detect the expression of β Ⅲ-tubulin regulated protein kinase,a neuronal marker as well as ERK and protein kinase B (Akt) phosphorylation.ResultsPC12 cell neurite formation rate was increased in a concentration dependent manner in the induction of DPA,increased by 2.4% (DPA 10 μg/ml,P>0.05),18.6% (DPA 30 μg/ml,P<0.05) and 25.0% (DPA 50 μg/ml,P<0.05) compared with that in the control group.DPA promoted the expression of β Ⅲ-tubulin (P<0.05) and the phosphorylation level of ERK and Akt (P<0.05,P<0.01).ConclusionDPA promotes PC12 cell neurites growth and its mechanism may be related to the activation of ERK and Akt signaling pathways.

Objective To study the clinical risk factors and the characteristics of cerebral magnetic resonance imaging (MRI) of hypoxic-ischemia encephalopathy (HIE) in full-term infants in high-altitude area.Method From January 2014 to December,2016 full-term Tibetan infants with HIE and healthy full-term Tibetan infants admitted to our hospital were enrolled in the study.General conditions and perinatal status were retrospectively analyzed.Univariate analysis and multivariate analysis were used to determine the risk factors of the HIE.MRI characteristics,location of the brain injuries and the correlation between HIE clinical grading and MRI grading were analyzed.Result During the study period,5 172 full-term Tibetan neonates were born in our hospital,198 of them were diagnosed of HIE and the incidence was 3.8％.According to HIE clinical grading,31 were mild,110 were moderate and 57 were severe.MRI grading included 34 mild,131 moderate and 33 severe.The main manifestations of MRI included white matter injury,especially subcortical white matter injury of frontal,parietal and occipital lobes,gray matter injury and diffuse cerebral edema.Mild HIE had a certain correlation with MRI grading,however,severe HIE had poor correlation with MRI grading.Multivariate Logistic regression analysis showed that abnormal birth weight (＜2 500 g or＞4 000 g) and intrauterine distress were independent risk factors of HIE in full-term Tibetan neonates[OR (95％ CI):3.663 (1.961～6.843) and 5.419 (2.487～11.807)].Conclusion Macrosomia at birth,low birth weight and intrauterine distress are independent risk factors of HIE in Tibetan full-term neonates in Lhasa.White matter injury is the main MRI manifestation of HIE.Clinical grading of Mild HIE has good consistency with the MRI grading,but MRI grading is milder than the clinical grading for those with moderate and severe HIE.

Objective To explore the therapeutic effects of soluble cytokines secreted by mesenchymal stem cells (MSCs) on acute liver failure (ALF).Methods MSCs isolated from Sprague-Dawley rats were determined by FACS analysis.Conditioned medium derived from MSCs (MSCs-CM) was collected and analyzed by a cytokine microarray.SD rats were divided into 3 groups:(1) ALF + dulbecco's modified eagle medium (DMEM) group:1 ml DMEM was injected into SD rats after D-Gal administration;(2) ALF + MSCs group:1 ml MSCs (1 × 106) was injected into SD rats after D-Gal administration;(3) ALF + MSCs-CM group:1 ml MSCs-CM was injected into SD rats after D-Gal administration.Biochemical indicators,survival rate,histology and inflammatory factors were studied.Exogenous recombinant rat IL-10,antirat IL-10 antibody and AG490 (STAT3 signaling pathway inhibitor) were administrated to explore the therapeutic mechanism of MSCs-CM.Results The respective serum biochemical indexes of ALF + DMEM group,ALF + MSCs group,and ALF + MSCs-CM group were:ALT (1 709.8 ± 372.1,865.5 ± 52.8,964.7 ± 414.6 U/L),AST (4234.0 ± 807.3,2440.8 ± 511.9,2739.8 ± 587.3 U/L),andTBil (79.3 ± 10.9,43.8 ± 7.0,61.2 ± 6.7 μg/L).The survival rates of the three groups were 10.0％,80.0％,and 70.0％,respectively.The levels of inflammatory factors in each group were IFN-γ (69.8 ± 4.7,46.4 ± 4.3,54.6 ± 2.4pg/ml),IL-1β (58.5 ± 7.6,40.5 ± 6.9,44.1 ± 6.0pg/ml),IL-6 (71.9 ± 16.1,38.4 ± 7.7,45.3 ± 9.0),and IL-10 (38.3 ± 6.0,75.4 ± 11.1,59.6 ± 11.9 pg/ml).Protein microarray results suggested that MSCs-CM expresses a variety of inflammatory-related cytokines,with IL-10 levels being most pronounced.IL-10 (ALT 1 126.9 ± 419.3 U/L,AST2370.8 ± 561.2 U/L) alone significantly reduced transaminase levels compared with ALF group (ALT 1 709.8 ± 372.1 U/L,AST 4234.0 ± 807.3 U/L),while anti-IL-10 antibody (ALT 1 568.5 ± 325.4 U/L,AST4043.7 ± 819.0 U/L) neutralized the therapeutic effect of MSCs-CM (ALT 964.7 ± 414.6 U/L,AST 2 739.8 ± 587.3 U/L).IL-10 could significantly increase the level of pSTAT3 in ALF rats (0.93 ± 0.03 vs 0.68 ± 0.01),while STAT3 inhibitor AG490 (0.84 ± 0.04) could decrease the expression of pSTAT3 and reverse the therapeutic effect of IL-10.Conclusion The factors released by MSCs,especially IL-10,have the potential therapeutic effect on ALF,and STAT3 signaling pathway may mediate the anti-inflammation effects of IL-10.

Objective To analyze the clinical features of patients with acute stage brucellosis in Shenzhen,and provide a scientific basis for prevention and control of brucellosis in immigrant city.Methods A retrospective analysis was conducted to collect clinical data of patients with brucellosis admitted to the Department of Infectious Diseases,Shenzhen People's Hospital from May 2013 to May 2018.The patient's epidemiology manifestations,pathogen and laboratory examination results,diagnosis and treatment outcomes and prognosis were analyzed.Results Among the 39 patients with brucellosis,males were predominant,with a male to female ratio of 1.4 ∶ 1.0 (23 ∶ 16),an age of (44.91 ± 17.18) years and 24 cases were non-Guangdong natives.There were 23 cases with epidemiological history,including 14 cases with mutton,sheep viscera and goat milk history;the disease occurred throughout the year,mainly from February to July,a total of 26 cases.The clinical manifestations of the patients were mainly fever,sweating,fatigue,joint and muscle pain,weight loss,and liver or spleen or lymph nodes swelling.The blood culture was identified as 38 cases of Brucella melitensis and 1 case of Brucella suis.All strains were sensitive to common antibiotics in vitro.All cases were diagnosed as acute stage of brucellosis,2 cases with orchitis,1 case with brucellosis meningoencephalitis,3 cases with spondylitis,and 3 cases with misdiagnosis.Thirty-nine patients were cured according to the "Brucellosis Diagnosis and Treatment Guidelines (Trial)" and were followed up for 1 year.Conclusions Patients with brucellosis in Shenzhen are mainly infected with Brucella melitensis;fever,sweating,joint and muscle pain are the main clinical symptoms;the patient's efficacy and prognosis are better after treatment;for the occurrence of occasional misdiagnosis,it is recommended that in immigrant cities,medical staff should strengthen their understanding of brucellosis.

Objective To evaluate the difference of set-up errors between thermoplastic mask and breast bracket in patients receiving intensity-modulated radiotherapy after breast conserving surgery,and the impact of clinical factors associated with set-up errors.Methods A total of 34 patients treated with intensity-modulated radiotherapy after breast conserving surgery from January 2016 to June 2018 were reviewed.Eighteen patients were fixed with thermoplastic mask,and sixteen were with breast bracket.Weekly CBCT scan records were used to analyze set-up errors,and group systematic and random errors were computed.The influence of clinical factors on set-up errors was also analyzed.Results The immobilization technique with thermoplastic mask showed great superior in comparison with breast bracket;however,only in the Ty(translation) and Ry(rotation),the differences had significance.Based on group systematic and random errors,PTV margins in Tx,Ty and Tzwere 2.65,4.36 and 2.87 mm in thermoplastic mask group,as well as 5.71,6.07 and 4.20 mm in breast bracket group,respectively.Multi-factor regression analysis showed that BMI was independent factors affecting set-up errors.Conclusions Compared with breast bracket,the immobilization technique with thermoplastic mask has the potential of reducing set-up errors and PTV margins in patients receiving intensity-modulated radiotherapy after breast conserving surgery,especially in patients with high BMI.

Objective:To analyze the influenza vaccination status of chronic disease management patients in Qingpu district of Shanghai and the vaccination characteristics of different characteristic populations, so as to provide scientific basis for improving the influenza vaccination rate of chronic disease patients in the community.Methods:By comparing the data of Shanghai chronic disease management information system, immunization planning information system and medical association platform, 89 453 subjects who met the inclusion and exclusion criteria in Qingpu district were selected as the research objects. The vaccination coverage rate of the study subjects was calculated according to gender, age group, urban and rural distribution, occupation, chronic disease type and quantity, and the vaccination coverage rate of different subgroups was compared to analyze the influencing factors of vaccination coverage rate.Results:Most of the 89 453 patients with chronic diseases were 60 years old and above (71.93%). Patients with hypertension, diabetes, chronic obstructive pulmoriary disease (COPD) and three chronic diseases accounted for 87.12%, 28.67%, 8.71% and 1.83%, respectively. Influenza vaccination coverage in the 2016/2017 flu season was low, at 0.32%. Influenza vaccination coverage rate of women (0.37%) was higher than that of men (0.27%), which was 1.41 times respectively(95% CI: 1.16, 1.72) that of men patients. The coverage rate of influenza vaccination for the 70-79 year-old group was the highest (0.74%), which was 1.74 times respectively(95% CI: 1.39, 2.19) that of 60-69 year-old patients. The vaccination coverage rate of government departments and institutions was the highest (1.14%), which was 12.58 times respectively(95% CI: 4.52, 34.99) that of retirees. The vaccination rate of COPD patients (3.68%) was 2.50 times (95% CI: 1.66, 3.77) higher than that of patients without COPD.Conclusions:Influenza vaccination rate for chronic disease management patients in Qingpu district of Shanghai is low. Gender, occupation, age and types of chronic diseases are the influencing factors. Patients with chronic disease management should be included in the priority vaccination targets for influenza vaccines, and vaccination intervention for occupational chronic diseases such as non-retired agriculture and forestry patients, especially male patients, should be strengthened to improve influenza vaccination coverage rate.

Objective:To investigate the expression of four cancer stem cell (CSC) markers (EpCAM, CD133, CD90 and CD24) in hepatocellular carcinoma tissues and peripheral blood circulating tumor cells (CTC),their value in the prognosis of patients with hepatocellular carcinoma.Methods:A total of 50 hepatocellular carcinoma tissues and 29 peripheral blood sample from 50 patients with hepatocellular cancer treated in Zhongshan Hospital Fudan University from October 2013 to September 2014 were collected and analyzed by flow cytometry or qRT-PCR to examine the expression of EpCAM, CD133, CD90 and CD24. The clinical data of patients were collected, including tumor size, tumor number, satellite lesions, vascular invasion, Edmondson stage, BCLC stage and liver cirrhosis, etc. The correlation between the expression of four markers in hepatocellular carcinoma tissues and CTC with the clinical data and survival time of patients were compared.Results:The positive expression rates of EpCAM, CD133, CD90 and CD24 in hepatocellular carcinoma tissues were 66% (33/50), 18% (9/50), 60% (30/50) and 56% (28/50); the positive expression rates in CTC were 55% (16/29), 38% (11/29), 31% (9/29) and 59% (17/29). CD90 expression in hepatocellular carcinoma tissue was positively correlated with the occurrence HCC liver cirrhosis ( P<0.05), while CD133 expression was negatively correlated with the 5-year survival rate of patients ( P<0.05). The expression of EpCAM and CD24 in peripheral blood CTC were closely related to the patient′s Edmondson stage ( P<0.05). The survival time of patients with CD133 positive expression in hepatocellular carcinoma tissue was lower than those without CD133 expression ( P<0.05); the survival rate of patients with EpCAM expressed in either tissue or peripheral blood CTC was lower than that of patients with EpCAM double negative expression ( P<0.05). The survival rate of patients with CD90 negative in HCC tissue and positive in peripheral blood was lower than that in patients with double negative/double positive in tissue and peripheral blood or patients positive in hepatocellular carcinoma tissue and negative in peripheral blood ( P<0.01). Conclusion:Different expression characteristics of four markers in cancer tissues and peripheral blood CTC might provide useful information about predicting prognosis of hepatocellular carcinoma. The expression of CD133 in tissues can be used as an important survival predictor of hepatocellular carcinoma patients. The differential expression of cancer markers in tissue samples and blood samples can provide more clinical prognostic information.

OBJECTIVETo understand the infantile nocturnal sleep-wake pattern developmental trajectory with Actiwatch, which would benefit the clinical assessment of infantile sleep.

METHODThis study was a longitudinal study conducted between 7 Oct, 2009-30 Oct, 2011 in 10 hospitals of 9 cities of China ( Beijing, Xi'an, Qingdao, Wuhan, Changsha, Chongqing, Huzhou, Xiamen and Liuzhou). Actiwatch was used to track the sleep-wake pattern development trajectory of healthy infants in the first year of life in the home setting. Participating infants were followed up at 10th day and 28th day during the first month, and then monthly from the second to the sixth month after birth, and then at ninth and twelve months of age respectively. Meanwhile, infantile sleep was observed continuously for about 60 hours at each visit. According to the characteristics of repeated measurement data of this study, two-level random effect model was adopted to analyze the trend of infantile nocturnal sleep-wake parameters changing with age, and the gender difference.

RESULTA total of 473 healthy infants were included in this study, among whom 246 (52.0%) were boys, and 227 (48.0%) were girls; 355 (75.1%) infants completed the whole year follow-up survey. With infants' age increasing, the latency of infants' nighttime sleep onset decreased from 66.8 minutes on 10th day to 15.5-18.7 minutes at 6-12 months of age. The number of night wakes also decreased with age, while uninterrupted sleep periods lengthened with age. On the 10th day, there were 3.0 times of nightwaking on average, and the longest continuous sleeping interval lasted for 227.6 minutes on average. At 12-month of age, infants could sleep continuously for 350.9 minutes at most on average, while the number of nightwaking decreased to 1.6 times per night on average. Generally, nighttime sleep efficiency increased from 66.3% on the 10th day to 86.3% at 12-month of age. The differences of sleep-wake patterns between boys and girls presented as boys' nocturnal longest uninterrupted sleep period was 19 minutes shorter(266.6 vs. 285.6 min), and the average nighttime sleep efficiency was 2.2% lower (74.2% vs. 76.4%) compared with girls respectively. And the differences of sleep efficiency between boys and girls reduced gradually along with the growth.

CONCLUSIONDuring the first 6 months after birth, infantile sleep-wake pattern undergo obvious change. The capability of sleep-onset and uninterrupted sleep improved with age, and the sleep efficiency increased.

China ; Female ; Humans ; Infant ; Infant, Newborn ; Longitudinal Studies ; Male ; Sleep ; physiology ; Surveys and Questionnaires