Objective To study the effect of low dose cyclophosphamide on function of CD4+CD25+Treg of mice with Lewis lung cancer and investigate the relationship between CD4+CD25+Treg and the genesis and development of tumor.Methods The models were divided into three groups:CTX group,tumor group and control group.In CTX group,models were established by injection CTX(25 mg?kg-1) and subcultivated Lewis lung cancer cells were injected subcutaneously to the right axilla of C57BL/6 mice 7 d later.The dynamic changes of tumor volume were observed.The changes of the number of CD4+CD25+Treg were detected by flow cytometer and the expression of Foxp3 mRNA in spleen was analyzed by semi-quantitative RT-PCR.The changes of T lymphocyte proliferation in spleen were detected by MTT test.The changes of T lymphocyte killing function in spleen were detected by LDH releasing test.Results Compared with tumor group,CTX treatment might delay the development of tumor in 19 d-mice with Lewis lung cancer.The number of CD4+CD25+ Treg in spleen of mice in CTX group was lower than that in tumor group(P0.05).Conclusion After CTX injection,the number of CD4+CD25+Treg and the expression of Foxp3 mRNA in spleen of mice with Lewis lung cancer decrease,the immunological function of mice with Lewis lung cancer increase.It can provid experiment evidence for the tumor immunotherapy aimed directly at CD4+CD25+Treg.

Objective:To evaluate the microtensile bond strength changes and patterns of fractures of the bonding interface after dentine surface treatment with carbodiimide-ethanol solution .Methods:1-eth-yl-3-(3-dimethylaminopropyl) carbodiimide (EDC) dissolved in ethanol was diluted into different con-centrations of 2, 1, 0.3, 0.1 and 0.01 mol/L EDC-ethanol solutions.Twenty-eight caries-free extracted human third molars were ground metallurgically to prepare flat occlusal mid-coronal dentin surfaces and etched with 35%( mass fraction) phosphoric acid gel .Then they were treated with EDC-ethanol solution for 60 s before the bonding procedure and randomly divided into five experimental groups corresponding to the tested EDC-ethanol concentrations .The ethanol treated and no pre-treated surfaces were used as con-trols.Single Bond 2 adhesive was applied and resin composite disk was stacked on the treated dentine surface .The teeth with resin composite disks were stored in water at room temperature for 24 h and then sectioned longitudinally to produce stick specimens for microtensile bond strength test .Fracture patterns were observed with a stereomicroscope .Results:The dentin surfaces pre-treated with 2 mol/L [ (22.17 ± 13.31) MPa] and 1 mol/L [(45.31 ±17.80) MPa] EDC-ethanol solutions resulted in statistically sig-nificant lower bond strength value ( P<0 .05 ) .Increasing numbers of fracture pattern at the resin-dentin interface were also found in this two groups with percentages of 81 .2%and 41 .3%respectively .No sig-nificant difference was observed in the groups with 0.3, 0.1, 0.01 mol/L EDC surface treatment ( P>0.05).Conclusion:No significant difference of immediate bond strengths was found in the 0.3, 0.1, 0 .01 mol/L groups compared with the control group .EDC-ethanol solution surface treatment with con-centrations of 2 mol/L and 1 mol/L resulted in decreasing of the bonding strength .

Objective To evaluate the clinical effect and safety of percutaneous closure of patent foramen ovalein treatment of migraine.Methods We studied 23 patients with migraine who were tested positive in Contrast Transcranial Doppler (TCDc) at the Department of Neurology in Bethune First Hospital of Jilin University between 2010 May and 2011 November.Procedures of transcatheterclosure of patent foramen ovale were performed at Department of Cardiovascular Surgery.At varied time points post-surgery,the patients were re-examined by TCDc and transthoracic echocardiograph (TTE) and followed up by phone interviewed with Headache Impact Test-6 (HIT-6) questionnaire to elevate improvement of migraine.The data was analyzed statistically.Results Percutaneous closure was successful in all patients without major long-term side-effects.At follow-ups at 48-542 days ((247 ± 145) days) post-surgery,the scores of HIT-6 were reduced to 38-70 (51 ± 9),compared to the pre-operative scores of 42-76 (61 ± 8).Disability by migraine had significantly improved (t =4.80,P ＜ 0.05).In 6 patients with migraine with aura,the scores of HIT-6 were reduced to 46-56 (51 ± 3),from preoperative scores of 63-76 (69 ± 4) and their disability affected by migraine had significantly improved (t =6.80,P ＜0.05).In 17 patients with migraine without aura,the scores of HIT-6 after surgery were reduced to 36-70 (52-10) from the preoperative scores of 42-69 (60 ± 8) and disability affected by migraine in this group had significantly improved (t =3.77,P ＜0.05).Among 18 patients having MRI or CT examination before the operation,8 patients had neurological events (such as cerebral infarction,cerebral ischemia) with average age of (44 ± 7) years old.In this group,disability affected by migraine had significantly improved (the HIT-6 scores:49 ± 7 vs 58 ± 10,t =2.49,P ＜ 0.05).Conclusion Percutaneous closure of PFO showed significant improvement in migraine treatment and it is a safe procedure.

Objective To compare the identifiability for depressive symptoms using different instruments while interviewing with different respondents in suicide prevention research in China. Methods One hundred and fifty-one suicide death cases (suicide group) and one hundred and twenty suicide attempt cases (attempt group) were recruited. For each identified cases, one family member proxy respondent, and another associate proxy respondent (friend or neighbor) and suicide attempter (only for attempt group) were interviewed separately by qualified psychiatrists. The Di-agnostic Screening Instrument for Depression (DSID) and the Structured Clinical Interview for DSM-Ⅳ Axis Ⅰ Disorders (SCID-Ⅰ) were administered to each respondent to identify the depressive symptoms based on diagnostic criteria for major depressive episode in DSM-Ⅳ. Data collected from family members and associate respondents were merged as proxy data. The concordances of the DSID and SCID-Ⅰfor identifying depressive symptoms, meeting for criteria of Major Depressive Episode (MDE) and Mild and Major Depressive Episode (MMDE), were calculated based on different respondents' data. The prevalence of depressive symptoms, MDE and MMDE, were compared among merged proxy data, family member respondent's data, and associate respondent's data in suicide group and attempt group, and between self-respondent's data and merged proxy data in suicide attempt group. Results In suicide group, based on merged proxy data, the prevalence of MDE was 41.1%(62 cases) for DSID and 41.7%(63 cases) for SCID-Ⅰ, and the Kappa coeffi-cient was 0.77. Based on suicide attempters' self-raported data, the prevalence of MDE was 23.7% (27 cases) and 22.0% (24 cases) for DSID and SCID-Ⅰ respectively, with a Kappa of 0.74. Based on merged proxy report in attempt group, 16 (13.3%) and 15 (12.5%) cases were met for criteria of MDE (Kappa=0.89), using the 2 instruments. In both of the suicide and attempt groups, the merged proxy data got higher prevalence of depressive symptoms, MDE and MMDE than that only based on family respondent's data or associate's respondent's data using both of the 2 instruments (all P<0.05). Compared with merged proxy data, attempters' self-reported data got higher prevalence of MMD and MMDE using both of the 2 instruments (all P<0.05). Conclusions Based on same respondent's data, SCID-Ⅰ performs as well as DSID in identifying depressive symptoms. Collecting data from 2 respondents would get higher prevalence of MDE or MMDE than only from one family member or one associate. In attempt group, the prevalence of MDE or MMDE based on merged proxy data were lower than that based on attempters' self-reported data.

Objective:To investigate the clinical manifestations and characteristics of tracheobronchial foreign bodies in children, and improve the diagnosis and treatment of foreign bodies.Methods:A retrospective analysis was performed on the clinical data of children with tracheobronchial foreign bodies confirmed by fiberoptic bronchoscopy between January 2010 to December 2019.The children with tracheobronchial foreign body who were treated in the Maternal and Child Health Hospital of Gansu Province.Results:A total of 967 cases were operated by soft electronic bronchoscope, and foreign bodies were removed by means of foreign body forceps or nets.Among them, 19 cases(3 cases with subglottic foreign bodies, one with row of pins, and the rest 15 cases with foreign bodies completely wrapped by granulation)were not removed, two cases were spontaneously coughed, and 946 cases (97.8%)were removed.Bronchial foreign bodies in children were more common in boys, with the ratio of male to female being 2.14∶1.The main age of onset was 1-3 years old(88.8%). The incidence was slightly higher in rural areas than that in urban areas(46.5% in urban areas, 53.5% in rural areas). Foreign bodies were inhaled most in March and least in June.From the perspective of season, winter and spring were more than summer and autumn.The foreign body types inhaled were mainly exogenous plant foreign body, accounting for 93.0%, among which peanut(31.7%)and melon seeds(19.2%)were the most common.The duration of foreign body inhalation was up to 347 cases(35.9%)in 1-3 days.There were 501 cases(51.8%)with endoscopic endobronchial inflammation, and 196 cases of children with varying degrees of granulomatous hyperplasia, accounting for 39.1% and 20.3% of the total.The foreign body in the right bronchus accounted for 50.0% and the left bronchus for 43.7%.There were 793 cases confirmed by imaging, with a positive rate of 81.9%, and 90.9% confirmed by CT.Conclusion:About 88.8% of tracheobronchial foreign bodies occurred in 1 to 3 years of age.The occurrence of foreign bodies had obvious gender, urban-rural and seasonal distribution characteristics, and more cases were male, in rural and winter as well as spring.

[ ABSTRACT] AIM:To explore the role of SHARPIN in regulation of Rip1 in castration-resistant prostate cancer LNCaP-AI cells.METHODS:The LNCaP-AI cells were treated with TNF-α+Z-VAD ( an inhibitor of pan-caspase) to activate necroptosis, which were compared to the cells treated with TNF-α+Z-VAD+Nec-1 ( an inhibitor of Rip1 ) .A blank group and a TNF-α-treated group were set up as controls.The cell viability in each group was measured by MTS as-say.In addition, SHARPIN was knocked down by siRNA, and the inhibitory efficiency was evaluated by RT-qPCR.The expression of Rip1 at mRNA and protein levels after knocking down SHARPIN was determined by RT-qPCR and Western blot to explore the underlying mechanism of regulatory network of necroptosis in prostate cancer.RESULTS: Compared with blank control group and TNF-α-treated group, the viability of LNCaP-AI cells treated with TNF-α+Z-VAD decreased by 28%(P<0.05).After treated with TNF-α+Z-VAD+Nec-1, the LNCaP-AI cells showed no significant difference in the viability compared with blank control and TNF-α-treated groups.Taken together, necroptosis may be an important way of cell death in LNCaP-AI cells.Besides, the expression of Rip1 at protein level was up-regulated following the inhibition of SHARPIN using siRNA, indicating that down-regulation of SHARPIN enhanced necroptosis via activating Rip1 in
LNCaP-AI cells.CONCLUSION:Necroptosis is an important way of cell death .Inhibition of oncogenic factor SHARPIN enhances necroptosis via activating Rip1 in LNCaP-AI cells.

[ ABSTRACT] AIM: To investigate the SALL4 expression, proliferation and apoptosis in the LNCaP cells after transfection of SALL4 siRNA.METHODS: The expression of SALL4 at mRNA and protein levels was detected by real-time PCR and Western blotting.MTS assay, colony formation assay and flow cytometry were used to determine the prolifer-ation, colony formation ability and apoptosis of the LNCaP cells.The effect of SALL4 on the expression of Bax and Bcl-2 was analyzed by Western blotting.RESULTS:Compared with negative control group, the expression of SALL4 at mRNA and protein levels in LNCaP cells was down-regulated by transfection of SALL4 siRNA ( P<0.05 ) .The proliferation rate and colony formation ability were decreased, while apoptosis rate increased in si-SALL4 group (P<0.05).Higher expres-sion of Bax and lower expression of Bcl-2 in si-SALL4 group were observed ( P<0.05 ) .CONCLUSION:Down-regula-tion of SALL4 by siRNA not only suppresses LNCaP cell proliferation and colony formation, but also inhibits Bcl-2 expres-sion and activates Bax expression to induce apoptosis.

Based on the traditional medical theory,the theory of"ear-brain-liver-spleen correlation"was proposed by linking the ear with the brain,liver and spleen.Then,by combining the Chinese medical etiology of Functional Dyspepsia(FD)and the corresponding modern medical mechanism,we realized that the close connection between the ear,brain,liver and spleen is the theoretical basis for treating FD from the ear.In recent years,a new type of vagus nerve stimulation therapy,auricular concha eletro-acupuncture(ACEA),has been developed to treat FD by regulating the liver and spleen through the regulation of the brain,relieving liver depression and strengthening spleen and qi,thus treating FD.In terms of modern medicine,this is related to its ability to reduce anxiety and depression,reduce pain perception,enhance gastrointestinal motility and reduce central and peripheral inflammation by regulating the activity of the corresponding brain functional areas.By exploring the application of auricular concha eletro-acupuncture in FD on the basis of the"ear-brain-liver-spleen correlation",we will better understand the unique role of auricular stimulation in regulating the brain and the blood of the liver and spleen,and promote the inheritance,innovation and development of TCM theory.

OBJECTIVETo investigate the presence of miRNA-144 in the saliva of patients with esophageal cancer and its value for early diagnosis of esophageal cancer.

METHODSSaliva samples were collected form patients with esophageal cancer admitted in the Fourth Affiliated Hospital of Jinan University and the First Affiliated Hospital of Guangzhou Medical College between January, 2011 and May, 2013, with saliva samples from 50 middle-aged healthy volunteers matched for age and gender ratio as the control group. The contents of miRNA-144 in the samples were detected with RT-PCR.

RESULTSThe levels of miRNA-144 in both the whole saliva and saliva supernatant were significantly higher in esophageal cancer group than in the control group (P<0.05). In the whole saliva, the cut-off point of miRNA-144 was ≥100, with a sensitivity of 74.6% and a specificity of 92.0% for esophageal cancer diagnosis (Az=0.865); in saliva supernatant, the cut-off point was ≥20 with a sensitivity of 53.7% and a specificity of 94.0% (Az=0.754), suggesting a moderate diagnostic value of miRNA-144 in whole saliva and saliva supernatant.

CONCLUSIONmiRNA-144 is highly expressed in the saliva of patients with esophageal cancer and can be used as a genetic marker for early diagnosis of esophageal cancer.

Biomarkers, Tumor ; Early Diagnosis ; Esophageal Neoplasms ; diagnosis ; Humans ; MicroRNAs ; analysis ; Middle Aged ; Saliva ; chemistry ; Sensitivity and Specificity

OBJECTIVETo investigate the effect of CD59 gene inhibition mediated by RNA interference on the proliferation and apoptosis of non-small cell lung cancer (NSCLC) GLC-P cells in vitro.

METHODSRecombinant plasmids for RNA interference of CD59 gene were constructed and transfected into GLC-P cells via lipofectamine 2000. The stably transfected cells were examined with real-time RT-PCR, MTT assay and enzyme-linked immunosorbent assay to investigate the changes in cell proliferation and apoptosis.

RESULTSCompared with the control cells, the cells transfected with CD59-siRNA showed significantly decreased expression levels of CD59 mRNA (P<0.05) and significantly inhibited cell proliferation.

CONCLUSIONCD59 gene is highly expressed in NSCLC and RNA interference-mediated CD59 silencing can strongly inhibit the proliferation and induce apoptosis in GLC-P cells, which shed light on a potentially new target for targeted gene therapy of NSCLC.

Apoptosis ; CD59 Antigens ; genetics ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Enzyme-Linked Immunosorbent Assay ; Genetic Therapy ; Humans ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Transfection