OBJECTIVETo study the molecular mechanism of Yangjing Zhongyu Decoction (YZD) n-butanol extracts (ZDC) and ethyl acetate extracts (YSYZ) in reducing androgen in porcine granulose cells by mitogen-activated protein kinase (MAPK) pathway.

METHODSPorcine granulose cells were isolated and cultured. They were inoculated by MAPK inhibitor PD98059 at different concentrations, and then they were divided into the blank control group (0), 1, 3, 10, and 25 micromol/L groups. After 24-h culture the cytochrome P450c17a (CYP17) mRNA expression level was detected using Real-time fluorescent quantitative PCR. Contents of androgen (testosterone) in the supernate were detected using RIA and optimal PD98059 concentration screened. After intervened by 10 micromol/L PD98059 for 24 h, the culture solution was intervened by effective ingredients of with or without YZD or YSYZ at various concentrations (0, 1 , 5, 25, 50 mg/mL) at various time points (3, 6, 18, 24 h). Expression levels of p-ERK1/2, c-Fos and CYP17 were detected by Western blot. Testosterone content in the supernate was determined by radioimmunoassay (RIA).

RESULTSTen pLmol/L PD98059 could obviously decrease p-ERK1/2 protein expression and increase CYP17 mRMA expression, and elevate testosterone content in the supernate (P < 0.05). ZDC and YSYZ at 25 ng/mL could increase p-ERK1/2 protein expression and c-Fos levels, and reduce CYP17 protein expression, and lower testosterone content in the supernate after 6-h intervention (P < 0.01).

CONCLUSIONEffective ingredients of YZD could reduce androgen production in porcine granulose cells through increasing activities of MAPK.

Androgens ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Flavonoids ; Granulosa Cells ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; RNA, Messenger ; Swine

AIM:To study the effect of epigallocatechin-3-gallate (EGCG) on the proliferation of human naso-pharyngeal carcinoma ( NPC) cells, and to explore its mechanism by targeting miR-34a.METHODS: Nasopharyngeal carcinoma CNE-2Z cells were treated with various concentrations of EGCG .The ability of cell proliferation was detected by CCK-8 assay, 5-ethynyl-2-deoxyuridine (EdU) incorporation assay and colony-forming assay.The cell cycle distributions were analyzed by flow cytometry .The protein levels of P53 and Notch1 were detected by Western blot .The expression of miR-34a and Notch1 mRNA was measured by real-time PCR.RESULTS:EGCG effectively inhibited the proliferation and colony formation of CNE-2Z cells in a dose-dependent manner , which was related to its induction of cell cycle arrest at G 0/G1 phase.The expression of P53 and miR-34a in CNE-2Z cells was significantly increased after treated with EGCG , while the expression of Notch1 at mRNA and protein levels was markedly suppressed .CONCLUSION:EGCG induces cell cycle arrest and suppresses cell proliferation by regulating the P 53/miR-34a/Notch1 pathway in NPC cells.

Objective: To observe the clinical efficacy of herb-partitioned spreading moxibustion at Baliao points plus climen for diminished ovarian reserve (DOR). Methods: A total of 60 patients with DOR were randomized into a spreading moxibustion group and a Western medicine group by the random number table method, with 30 cases in each group. The Western medicine group was treated with climen, starting from the 5th day of the menstrual cycle for 21 d. The spreading moxibustion group was treated with herb-partitioned spreading moxibustion at Baliao points on the basis of the medication in the Western medicine group, 1 h per time, once a week. The treatment was performed for 1 month as one treatment course in both groups, for 3 courses in total. The serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) in the patients were measured before and after treatment. The peak systolic velocity (PSV) and resistance index (RI) were also detected. The traditional Chinese medicine (TCM) symptom score was evaluated. The clinical efficacy was evaluated after treatment. Results: The total effective rate in the spreading moxibustion group was 93.3％, which was significantly higher than 80.0％ in the Western medicine group, and the difference between the groups was statistically significant (P<0.05). After treatment, the TCM symptom scores, the serum FSH levels, FSH/LH ratios and RI in both groups decreased, and the intra-group differences were all statistically significant (all P<0.05). The serum E2 level and PSV increased compared with those in the same group before treatment, and the intra-group differences were statistically significant (all P<0.05). After treatment, the TCM symptom score, the serum FSH level, FSH/LH ratio and RI in the spreading moxibustion group were lower than those in the Western medicine group, while the serum E2 level and PSV were higher than those in the Western medicine group, and the differences between the groups were statistically significant (all P<0.05). Conclusion: Herb-partitioned spreading moxibustion at Baliao points plus climen can produce valid therapeutic efficacy for DOR. It can improve the clinical symptoms, regulate serum hormone levels and increase ovarian blood perfusion, thus improving ovarian reserve function, producing more significant efficacy than climen alone.

OBJECTIVETo investigate the common causes of recurrent wheezing in young children.

METHODSElectronic bronchoscopy was performed on 67 children with recurrent wheezing or who did not respond to the conventional treatment.

RESULTSThe electronic bronchoscopy showed intimitis in trachea and bronchi in 19 cases, intimitis and inflammatory stricture in 11 cases, foreign bodies in the bronchi in 11 cases, trachea and bronchus softening in 19 cases, and bronchopulmonary dysplasia in 3 cases. The other 4 cases presented endometrial tuberculosis, epiglottic cyst, laryngeal papilloma or compression outside trachea (thymus) under the electronic bronchoscope.

CONCLUSIONSIn addition to inflammation, trachea and bronchus softening as well as foreign bodies in the bronchi are also the common causes in children with recurrent wheezing or who do not respond to the conventional treatment. Electronic bronchoscopy appears to be an effective way to determine the cause in these children.

Bronchoscopy ; methods ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Recurrence ; Respiratory Sounds ; etiology

Objective To investigate the status of control of endemic fluorosis in Liaoning Province.Methods To investigate the prevalence rate of endemic fluorosis and water fluoride content in regions with different extent of endemic fluorosis,dental fluomsis among 8-12 years old children and clinical fluorosis at adult above 16 years old were extensively surveyed,urinary fluoride among 8-12 years old children was detected.Results We surveyed 842 undefluorided drinking water in endemic fluorosis villages and 1234 projects of improving drinking water in 1829 endemic fluorosis viflages.Water fluoride content was 0.01-7.10 mg/L in unimproved drinking water in endemic fluorosis resions,averaging(0.96±0.64)mg/L;29.2%(246/842)of the endemic fluorosis regions had a fluoride content more than 1.2 mg/L In 1234 projects of improving drinking water.drinking water fluoride content was between 0.06-7.67 mg/L.The project normally operated and having a fluoride content≤1.2 mg/L accounted for 68.31%(843/1234),while 31.69%(391/1234)of the projects did not function well.The prevalence of dental fluomsis in 8-12 years old child ren in endemic fluorosis regions was 24.4%(2960/12 127),the prevalence of clinical fluorosis among adults was 2.22%(1900/85 636).The prevalente of dental fluorosis in slight,moderate and serious fluorosis regions had remarkable statistics differences(X2＝19.25,P＜0.01).The prevalence of dental fluorosis of children in serious fluorosis regions was the highest,reaching 100%,while the prevalence of skeletal fluorosis wns 18.03%(97/538).The median of urinary fluoride was 2.01.2.00mg/L in serious and slight fluomsis regions,respectively.Conclusions Endemic fluorosis is still serious.so we need urgently to improve water in serious fluorosis regions without defluoridaton of drinking water.Endemic fluomsis resions where worn-out and closed defluoriding projects exist need defluoriding management.

BACKGROUNDPulmonary embolism (PE) is rare and seldom considered in adolescent patients; however it occurs with a greater frequency than is generally recognized, and it is a potentially fatal condition. The aim of the current study was to understand its epidemiology, clinical features and the cause of delay of its diagnosis in adolescents.

METHODSA retrospective analysis of nine adolescents with acute PE admitted to the Peking University Third Hospital over the past 16-year period was performed. The epidemiology, clinical features and risk factors of the adolescents were described and compared with those of adults and elderly patients. The time to diagnosis and misdiagnosed diseases were analyzed. Pretest probability of PE was assessed retrospectively by the Wells score and revised Geneva score.

RESULTSThe incidence of PE was 43.6 per 100 000 hospitalized adolescents in our hospital. The incidence of PE in adolescents was much lower than that in adults and PE is diagnosed in about 1/50 of elderly people. The clinical features in adolescents were similar to those in adults. But fever and chest pain were more common in adolescents (P < 0.05). The major risk factors included surgery, systemic lupus erythematosus (SLE), thrombocytopenia, long-term oral glucocorticoids and trauma. The mean diagnostic time was (7.8 ± 8.4) days. Six cases had a delayed diagnosis. The mean delay time from symptom onset to diagnosis was (11.0 ± 8.8) days. The time of presentation to diagnosis in patients initially admitted to the emergency department was less than one day, and was much shorter than the time in outpatients, (9.4 ± 7.5) days. Most of the patients were initially misdiagnosed with a respiratory tract infection. Most patients' values of Wells score or revised Geneva score were in the moderate or high clinical probability categories; 88% by Well score vs. 100% by revised Geneva score.

CONCLUSIONSPE was seldom considered in the adolescent patients by physicians, especially outpatient physicians, so the diagnosis was often delayed. If adolescent patients complain of dyspnea or chest pain or syncope with/without fever, and they had risk factors such as surgery, thrombocytopenia and trauma, PE should be considered and included in the differential diagnosis.

Adolescent ; Adult ; Diagnosis, Differential ; Diagnostic Errors ; Humans ; Male ; Probability ; Pulmonary Embolism ; diagnosis ; epidemiology ; etiology ; Retrospective Studies ; Risk Factors

OBJECTIVETo detect plasma concentrations of vascular endothelial cell growth factor (VEGF) and tissue factor (TF) in children with acute lymphoblastic leukemia (ALL) and explore their clinical significance in ALL.

METHODSThirty-three children with newly diagnosed ALL, including 18 cases of low risk, 7 cases of moderate risk and 8 cases of high risk, were enrolled in this study. Twenty-five patients received a complete remission and 8 cases were in non-remission after conventional remission induction chemotherapy. Plasma concentrations of VEGF and TF in the patients were detected using ELISA before and after treatment. Sixteen healthy children served as normal control group.

RESULTSPlasma concentrations of VEGF and TF in ALL patients before treatment were significantly higher than those in normal controls (P < 0.01). Plasma concentrations of VEGF and TF in the non-remission group before treatment were significantly higher than those in the remission group (P < 0.05) and the control group (P < 0.01). After treatment the plasma concentrations of VEGF and TF in the non-remission group were not significantly reduced and higher than those in the remission and the control groups (P < 0.01). There were significant differences in plasma concentrations of VEGF and TF among the low-risk, moderate-risk and high-risk groups before and after treatment (P < 0.05). Plasma concentrations of VEGF and TF in the high risk group were not significantly reduced after treatment and higher than those in the control group (P < 0.01). A linear correlation was noted between plasma VEGF and TF concentrations in ALL patients before treatment (r=0.50, P < 0.01).

CONCLUSIONSVEGF and TF play an important role in the development of ALL and may be useful to the evaluation of the severity and the outcome in ALL.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; etiology ; Thromboplastin ; analysis ; Vascular Endothelial Growth Factor A ; blood

[Absract] Objective This paper was designed to reveal the new mechanism on ASI Ⅱ triggered CD4+T cells activation via regulating CD45 molecular and provide a basis for the theoretical foundation of antitumor immunotherapy of Astragalus.Methods The CD4+T cells were randomly divided into negative group,stimulated control group,ASI Ⅱ group,CD45 inhibitor group,and the combination of ASI Ⅱ and CD45 inhibitor group.Besides negative group,the cells from other groups were activated by anti-CD3/CD28 antibody.ASI Ⅱ group was treated with 10 nmol/L ASI Ⅱ,CD45 inhibitor group was treated with 0.8 μmol/L CD45 inhibitor,and the combination group were treated with 10 nmol/L ASI Ⅱ and 0.8 iμmol/L CD45 inhibitor.After 36h culture,the proliferation of CD4+T cells was detected by Ki-67 intracellular staining assay.Cytokine production of Th1 and Th2 were examined ELISA method.The proportion of surface marker (CD44 and CD25)and Th1 intracellular cytokines (IFN-γ) were detected by flow cytometry.Results Compared with stimulated group,Astragaloside Ⅱ group in CD4+Ki67+T positive proportion (5.37％ ± 0.92％ vs.1.19％ ± 0.23％),in CD4+CD25+ positive proportion (50.23％ ± 4.65 ％ vs.15.89％ ± 1.13％),in CD4+CD44+ positive proportion (33.16％ ± 6.08％ vs.15.36％ 4 1.45％),in CD4+IFN-γ+ positive proportion (1.42％ ± 0.44 ％ vs.0.38％ ± 0.06％) were significntly increased.And the secretion of IFN-γ,IL-4 and IL-2 in ASI Ⅱ group were higher than stimulated group.The anti-mouse CD45 Ab treatment markedly blocked the proliferation and Th1 cytokines production which induced by ASI Ⅱ.Furthermore,the anti-mouse CD45 Ab treatment significantly decreased the expression of surface marker (CD44 and CD25).Conclusions Activating CD45 protein tyrosine phosphatase may be involved in ASI Ⅱ triggered CD4+T cells activation.This study will provide a basis for antitumor immunotherapy of Astragalus.

OBJECTIVETo study the effects of minimal residual disease (MRD) level on day 33 of remission induction and IKZF1 genotype on the survival of children with B-lineage acute lymphoblastic leukemia (B-ALL).

METHODSA total of 152 children with newly-diagnosed B-ALL who had complete remission after the first cycle of the chemotherapy and had complete follow-up information were enrolled in this study. According to the MRD detection by flow cytometry on day 33 of remission induction, they were divided into three groups: standard-risk (SR) group (MRD <10; n=60), intermediate-risk (IR) group (10≤ MRD <10; n=55), and high-risk (HR) group (MRD ≥10; n=37). Nested RT-PCR was used to determine the IKZF1 genotype of all children before chemotherapy. The effects of MRD level on day 33 of remission induction and IKZF1 genotype on the recurrence-free survival (RFS) of children with B-ALL were analyzed.

RESULTSThere were 7 common IKZF1 subtypes in all the 152 children with B-ALL: IK1, IK2/3, IK4, IK6, IK8, IK9, and IK10. Of the 152 children, 130 had functional subtypes of IKZF1 and 22 had non-functional subtypes of IKZF1. During the follow-up period, relapse occurred in 26 (17%) children, and the recurrence rate was highest in the HR group (P<0.05). However, there was no significant difference in the recurrence rate between the SR group and the IR group (P>0.05). The cumulative recurrence rate of the children with non-functional subtypes of IKZF1 was significantly higher than that of those with functional types of IKZF1 (P<0.01). The predicted 5-year RFS rates in the SR, IR, and HR groups were (94.2±2.9)%, (86.7±3.8)%, and (56.2±4.5)% respectively (P<0.05). The 5-year RFS rate of the children with functional subtypes of IKZF1 was significantly higher than that of those with non-functional subtypes of IKZF1 (P<0.01). There was no significant difference in the predicted 5-year RFS rate between the children with functional subtypes of IKZF1 and those with non-functional subtypes of IKZF1 in the SR group (P>0.05). However, the predicted 5-year RFS rate of the children with functional subtypes of IKZF1 was significantly higher than that of those with non-functional subtypes of IKZF1 in the IR group and the HR group (P<0.05).

CONCLUSIONSB-ALL children with non-functional subtypes of IKZF1 have a high recurrence rate, and the recurrence rate will be even higher in B-ALL children with non-functional subtypes of IKZF1 and MRD ≥10 on day 33 of chemotherapy.

Antineoplastic Combined Chemotherapy Protocols ; Child ; Child, Preschool ; Female ; Genotype ; Humans ; Ikaros Transcription Factor ; genetics ; Male ; Neoplasm, Residual ; genetics ; mortality ; therapy ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; mortality ; therapy ; Prognosis ; Recurrence ; Remission Induction ; Survival

BackgroundAn ideal animal model is very important for the investigation of the immune mechanism of high-risk rejection corneal transplantation.ObjectiveThis study was to compare three methods of creating a high-risk corneal transplantation model in rabbits to study high-risk rejection corneal transplantation.MethodsForty-five New Zealand white rabbits were utilized and assigned randomly to three groups of different modeling methods,with 15 rabbits for each group.The high-risk corneal transplantation models were created by suturing with 5-0 silk thread in 4 quadrants,inducing alkali burn with 1 mol/L NaOH or corneal xenotransplantation.In the suturing group and alkali burning group,the rabbits received a unilateral 7.25 mm diameter corneal allograft after corneal neovascularization was induced,and in the xenotransplantation group,corneas from cats were used as donors.Rabbits were followed-up for 4 weeks in all groups.Corneal neovascular area was calculated and compared among the three groups.The amount of rejection,inflammatory index ( IF),neovascularization and histology of grafts were clinically scored to calculate the reject index (RI).ResultsThere were 14,15 and 15 rabbits that survived the high-risk penetrating corneal transplantation,respectively,in the suturing group,alkali burning group and xenotransplantation group.Two weeks after operation,the IF scores were 0.543 ± 0.103,0.811 ± 0.054 and 0.191 ±0.087,and the RI were 2.111±0.928,7.0±0.816 and 3.182±0.751 in the suturing group,alkali burning group and xenotransplantationgroup,respectively,showingstatisticallysignificantdifferencesamongthethreegroups (x2 =25.736,22.432,P =0.000).The IF value was lower in the xenotransplantation group compared with the suturing group and alkali burning group (Z =3.841,3.993,P =0.000),and that of the suturing group was lower than the alkali burning group (Z =3.568,P =0.000).The RI value of the xenotransplantation group was significantly raised in comparison with the suturing group and declined in comparison with the alkali burning group (Z =2.373,P =0.018;Z =3.936,P =0.000),and that of the suturing group was lower than the alkali burning group (Z =3.729,P =0.000 ).The survival times of the grafts were ( 17.9±2.0 ) days,( 13.4 ±2.4) days and ( 15.5 ±2.0 ) days in these three groups with a significant difference among them ( F =9.474,P =0.001 ).The neovascularization area in the xenotransplantation group was smaller than the suturing group and alkali burning group (P＜ 0.05 ).Histological examination revealed a large number of inflammatory cells infiltration in the grafts 2 and 4 weeks after transplantation in the suturing group and alkali burning group,but less inflammatory cells were seen in the xenotransplantaion group.Immunofluorescence staining showed abundant CD4+ T positive cells in the grafts in the three groups.Conclusions The cat-rabbit corneal xenotransplantation can induce stable and moderate immune rejection.This animal model has milder inflammatory response and less corneal neovascularization than the suture and alkali burn models.This method therefore is an ideal model for high-risk corneal transplantation.