Objective To study the effect of a new immunomodulator composed of muramyl dipeptide(MDP) and anti-CD10monoclonal antibody(MDP-Ab)on the dendritic cells(DC)of children with acute leukemia. Methods DC was adopted to divide the children with acute lymphoblastic leukemia into 6 groups,including the control group,unconjugated anti-CD10alone,unconjugated MDP alone,MDP-Ab alone,lipopolysaccharide(LPS)alone and MDP-Ab + LPS.The immunophenotypes,the endocytosis interleukin-12(IL-12)were detected.The stimulation index of autologous lymphocytes was assayed by adopting 5-(and 6)-carboxyfluorescein diacetate,succinimidyl es-ter(CFSE)-staining method.The supernatants of DC and autologous lymphocytes were used to detect the level of in-terferon-γ(IFN-γ)by using enzyme-linked immunosorbent assay.Results (1)DC immunophenotype:The ex-pressions of human leukocyte antigen-DR(HLA-DR),mature molecule(CD83)and co-stimulatory molecules (CD80and CD86)were increased significantly upon DC triggered with MDP-Ab,compared with the control group,un-conjugated anti-CD10group,and unconjugated MDP group,but lower than those in LPS and combination of MDP-Ab with LPS(F=629.62,P=0.000).(2)The level of IL-12:a significant increase in IL-12 level was detected in MDP-Ab group,LPS group,and combination of MDP-Ab with LPS group,compared with the control group,uncon-jugated anti-CD10group,and unconjugated MDP group(F=857.87,P=0.000). There were significant differences among the first three groups.(3)Endocytosis assay:The uptake of DCs stimulated by unconjugated anti-CD10,un-conjugated MDP,MDP-Ab immunoconjugate,LPS or combination was lower than that of immature DC in the control group which was(81.3 ± 10.1)%.(4)Mixed lymphocyte reaction and IFN-γ level:DC,treated with MDP-Ab, LPS and combination,stimulated more CFSE positive cells and higher level of IFN-γ secretion than the control group and unconjugated anti-CD10group,unconjugated MDP group. The most significance was observed in combination of MDP-Ab with LPS(F=393.36,P=0.000;F=2 497.18,P=0.000).Conclusion It is concluded that MDP-Ab could promote the proliferation and maturation of DC derived from blood of children with acute leukemia.

OBJECTIVE: To compare the differences in the clinical therapeutic effects on perimenopausal syndrome (PMS) between the combined treatment with the transcutaneous electrostimulation at Shuitu (ST 10) and the seed-pressure therapy at the auricular points and the hormone replacement therapy. METHODS: A total of 64 PMS patients were divided into an observation group (30 cases) and a control group (34 cases) according to patient's willingness. In the observation group, the transcutaneous electrostimulation at Shuitu (ST 10) was combined with the seed-pressure therapy at the auricular points. The stimulation intensity at Shuitu (ST 10) was ranged from 15 to 20 mA, for 20 min in each treatment, twice a week. The 8 treatments made one course and 3 courses were required. Additionally, the seed-pressure therapy was used at neifenmi (CO), Shenmen (TF), luanchao, zigong, gan (CO), shen (CO), once a week, retained for 5 days. The 4-week treatment was as one course and a total of 3 courses were required. In the control group, the hormone replacement therapy was applied. On Day 5 of menstruation, progynova was prescribed for oral administration, 1 mg, once a day, continuously for 21 days. On Day 12 in the oral administration of progynova, dydrogesterone was used, 20 mg, once every day, continuously for 10 days. Totally, 3 courses were required. Separately, before treatment and in 1 and 3 months after treatment, Kuppermann score, the sexual hormone levels [follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E)] and the adverse reactions were observed in the patients of the two groups. RESULTS: In 1 and 3 months after treatment, Kuppermann scores were reduced significantly as compared with those before treatment in the two groups (all <0.05). The levels of serum FSH and LH were reduced remarkably as compared with those before treatment in the two groups (all <0.05) and the level of serum E was increased remarkably as compared with that before treatment in the two groups (all <0.05). After treatment, the differences in the above indexes were not significant between the two groups (all >0.05). In the control group, the incidences of the adverse reactions such as vaginal bleeding, bread distending pain and endometrial thickening were higher than those in the observation group [17.6% (6/34) vs 0% (0/30), 20.6% (7/34) vs 3.3% (1/30), 8.8% (3/34) vs 0% (0/30), all <0.05]. CONCLUSION The combined treatment with the transcutaneous electrostimulation at Shuitu (ST 10) and the seed-pressure therapy at the auricular points achieves the similar therapeutic effects on PMS as compared with the hormone replacement therapy. This combined therapy effectively relieves the clinical symptoms and improves the sexual hormone levels. The adverse reactions and the complications are less obviously as compared with the hormone replacement therapy.
Acupuncture Points ; Acupuncture, Ear ; Female ; Follicle Stimulating Hormone ; Humans ; Luteinizing Hormone ; Perimenopause

Objective To investigate the correlation between cytochrome P450(CYP) 2C19 gene polymorphism with major adverse cardiovascular events(MACE) after PCI in the patients with coronary heart disease(CHD).Methods A total of 233 patients with CHD undergoing PCI in the cardiology department of our hospital from January 2014 to January 2015 were selected.All patients were given the standardized dual anti-platelet therapy of aspirin and clopidogrel.The occurrence situation of MACE within 1 year(unstable angina pectoris,cardiac death,in-stent restenosis,non-fatal myocardial infarction) was obtained by follow up.All patients were divided into the MACE group and non-MACE group.The PCR solubility curve was adopted to detect the CYP2C19 gene polymorphism.Results Among 233 cases of CHD,37 cases (15.88％) developed cardiovascular events and 196 cases (84.12 ％) did not develop vascular events;the age,sex,hypertension and diabetes mellitus had no statistical differences between the two groups(P＞0.05).The frequency of CYP2C19 * 1 in the included cases was 68.45％,which of CYP2C19 * 2 was 28.33％ and which of CYP2C19 * 3 was 3.22％.The extensive-metabolism,intermediate metabolism and slow metabolism types in the cardiovascular events group accounted for 5.41 ％,64.86 ％ and 29.73 ％ respectively,while which in the non event group were 59.69 ％,29.08％ and 11.22％ respectively,the CYP2C19 genotype distribution had statistically significant difference between the two groups(P＜0.05).The multivariate Logistic regression analysis showed that CYP2C19 intermediate metabolism type [OR 2.562,95％CI(2.825,7.350),P=0.021 0],slow metabolism type [OR 5.139,95％CI(1.289,5.232),P＜0.01],hypertension [OR 2.480,95 ％CI(1.079,5.698),P=0.032 4],smoking[OR 4.802,95％CI(1.082,18.371),P=0.029 0] were the independent risk factors for the occurrence of cardiovascular events in the patients with CHD.Conclusion CYP2C19 * 2 and CYP2C19 * 3 gene polymorphism are the independent risk factors for MACE occurrence after PCI in the patients with CHD.

Objective To establish oxidative stress model of hydrogen peroxide treatment by using human umbilical vein endothelial cells (ECS) as cell model to study the protective mechanism of anti oxidative stress and determine the signal transduction pathway of remifentanil.Methods Primary cultured human umbilical vein endothelial cells which were incubated with 0.1 M hydrogen peroxide to establish injury model to study remifentanil protection and related pathways.The experiment was divided into nine groups: control group (group C), Hydrogen peroxide group (group H1), Hydrogen peroxide+SP600125 group (group H2), Hydrogen peroxide+SB203580 group (group H3), Hydrogen peroxide+PD98059 group (group H4), hydrogen peroxide+remifentanil group (group HR1), hydrogen peroxide+remifentanil+SP600125 group (group HR2), hydrogen peroxide+remifentanil+SB203580 group (group HR3), hydrogen peroxide+remifentanil+PD98059 group (group HR4).Groups H1, H2, H3 and H4 only performed MAPK pathway blockade experiments.Groups HR1, HR2, HR3 and HR4 individually added remifentanil 10 ng/ml to protect 1 h.SOD activity, MDA level, Caspase-3 activity were detected and anti oxidative stress of remifentanil observed to confirm preliminary transduction pathway;Using RT-PCR expression levels of c-Jun before was observed before and after treated with remifentail 10 ng/ml.The aim was to determine the transduction pathway of the signaling molecules.Results Compared with group C, SOD activity were decreased significantly, MDA performance level were increased significantly in groups H1, H2, H3 and H4 (P<0.05).Compared with group H1, SOD activity was increased significantly, MDA performance level was decreased significantly in group HR1 (P<0.05).SOD activity difference and MDA performance level of groups HR2 and H2 had no statistical significance.Compared with group H3, SOD activity was increased significantly, MDA performance level was decreased siginificantly in group HR3 (P<0.05).Compared with group H4, SOD activity was increased significantly, MDA performance level was decreased significantly in group HR4 (P<0.05).Caspase-3 activity of groups H1, H2, H3 and H4 were higher significantly than that of group C (P<0.05).The level of C-Jun mRNA in group H1 was significantly higher than that of group C;But it was higher in group HR1 than that of group C, it was significantly lower than that of group H1 (P<0.05).Conclusion By activating the JNK pathway and its downstream signaling molecule c-Jun, remifentanil 10 ng/ml has the effect of increasing SOD activity, reducing the level of MDA expression and playing a role in anti oxidative stress.

ObjectiveTo evaluate the effects of diagnosis of cervical insufficiency and different management on pregnancy outcomes.MethodsFrom June 2004 to May 2010, a retrospective analysis was carried out on 554 patients with cervical insufficiency in nine hospitals in Guangdong Province, China. The patients were divided into two groups, the cervical cerclage surgical treatment group (surgical group,n=357) and the expectant treatment group (n=197). These patients were then re-diagnosed according to the unified diagnostic criteria by the American College of Obstetricians and Gynecologists (2014), and divided into the definite diagnosis group (n=425) and the indefinite diagnosis group (n=129). The two independent samplest-test andChi-square test were used to compare pregnancy outcomes between the definite and indefinite diagnosis groups, and the different treatment groups.ResultsThe full-term delivery rate in the surgical group was significantly higher than that in the expectant treatment group [40.3% (144/357) vs 23.4% (46/197),χ2=16.254, P=0.000], and the late abortion rate was lower in the surgical group than in the expectant treatment group [22.4%(80/357) vs 40.1% (79/197),χ2=19.419,P=0.000]. In women with a definite diagnosis of cervical insufficiency, full-term delivery rate [44.7% (117/262) vs 20.9% (34/163),χ2=24.844,P=0.000], and newborn body weight were significantly higher in the surgical group [(2 664.3±762.2) vs (2 416.9±845.0) g,t=1.160,P=0.014] than in the expectant treatment group and the late abortion rate was significantly lower [21.4% (56/262) vs 41.1% (67/163),χ2=19.021,P=0.000]. Cervical cerclage in the indefinite diagnosis group did not resulted in raising the full-term delivery rate [28.4% (27/95) vs 35.3% (12/34),χ2=0.561], preterm delivery rate [46.3%(44/95) vs 29.4% (10/34),χ2=2.940], late abortion rate [25.3% (24/95) vs 35.3% (12/34),χ2=1.252] and newborn body weight [(2 526.5±761.8) vs (2 683.4±725.8) g,t=0.004] compared with expectant treatment group (allP>0.05). Pregnancy outcomes in the surgical treatment group in relation to twin pregnancies were not significantly different in the≥28 weeks delivery rate [81.4% (37/46) vs 69.2% (18/26),χ2=1.156], late abortion rate [19.6% (9/46) vs 30.8% (8/26),χ2=1.156] and newborn birth weight [(2 003.2±621.0) vs (1 807.5±609.4) g, t=0.057] compared with those in the expectant treatment group (allP>0.05).ConclusionsIn accordance with the diagnostic criteria for cervical insufficiency and indications for cervical cerclage in surgical cases, cervical cerclage can effectively improve pregnancy outcome. But cervical cerclage is not recommended in twin pregnancies with cervical insufficiency.

Objective To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. Methods (1) Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis (VVC) murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group (n=15) and control group (n=15) . Suspension(30μl, 100μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline (PBS) was given to the control group. (2) Tweenty-four hours after treatment, the fungal burden and colony-forming unit (CFU) of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. Results (1) VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group. (2) Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×104 CFU/ml] than that in the control group [(8.5 ± 2.1) × 104 CFU/ml, P=0.017]. IFN-γlevel of the test group was increased [(257 ± 11) vs (197 ± 4) pg/ml, P=0.000], while the level of IL-10 was reduced [ (287 ± 15) vs (379 ± 17) pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group (0.892±0.008 vs 0.496±0.013, P=0.000). Conclusion Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γand IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.

Objective Tracking of the vaginal microflora recovery and the expression of immune factors from untreated and treated patients with bacterial vaginosis (BV) by using different administration regimens and studying the relationship of treatment results and regimen selections. Methods 25 healthy females were selected as a control group and 100 BV patients were randomly divided into 4 groups (n=25/group). Group A: Intravaginal administration of metronidazole (× 7 d), Group B:Continuous intravaginal administration of metronidazole (× 7 d) and then live Lactobacillus Capsule (× 7 d) , Group C: Intravaginal administration of nifuratel (× 7 d), Group D: Continuous intravaginal administration of nifuratel (× 7 d) and then live Lactobacillus Capsule (×7 d). The microecological assessment system and EILSA were used to compare the clinical efficacy, vaginal microflora recovery and the changes in IL-8, TLR2 and TNF-αof the vaginal lavage fluid in healthy women or patients with bacterial vaginosis before and after treatments by four treatment strategies. Results ① The vaginal microflora imbalance, flora disturbance, pH value increased were presented in BV group compared with the control group.②Compared to the median of IL-8, TLR2 and TNF-α in vaginal lavage fluids of control group, there was no significant difference in IL-8 level but both TLR2 and TNF-αwere significantly increased (P<0.05) in BV group. The immune factors had no significantly difference in all BV groups.③The therapeutic effect in each BV groups was compared after stopping treatment for 7 days. The cure rate and the vaginal microflora recovery rate were significant higher in group B and D than group A and C (P<0.05). ④ After treatment there was no significant change in IL-8 level but there was an obviouslydecrease in TLR2 and TNF-α(P<0.05). The decreased levels are more significant in groups B and D than groups A and C (P<0.05). Conclusion By combining with the microecological assessment system to evaluate the therapeutic effect of BV, our research suggests that the sequence schemes of nifuratel plus live Lactobacillus Capsule is more effective in therapy effect, restoring normal vaginal micro-ecological environment and vaginal local immunity than metronidazole used alone.

Objective To study the inhibitory effect of antimicrobial peptide LL-37 on Candida albicans through its ability to promote the secretion of immune factors by vaginal epithelial cells. Methods (1) LL-37 prokaryotic expression vector pET-Duet/LL-37 was constructed and its expression was induced in Escherichia coli M15. The expressed LL-37 fusion protein was purified and identified by western blot. Antifungal activity of the purified protein was initially identified by Kirby-Bauer (K-B) method. (2) Purified LL-37 protein was added to human vaginal epithelial cells co-cultured with Candida, and inhibitory effect on Candida growth was determined by the glucose consumption method. Interferon γ (IFN-γ), interleukin 10 (IL-10) concentration and IFN-γ/IL-10 ratio were measured by ELISA at different time points. Results (1) LL-37 fusion protein was purified to 96% purity at a concentration of 433.92 μg/ml, and was shown to possess anti-fungal activity confirmed by the K-B method. (2) A Candida-vaginal epithelial cells co-culture system was successfully constructed. LL-37 recombinant protein inhibited the growth of Candida with absorbance values significantly higher in the treatment group compared to the control group at all measured time points (12-hour:3.008±0.003 versus 2.967±0.003, 24-hour:2.941±0.003 versus 2.601±0.003, 48-hour:2.893 ± 0.004 versus 2.409 ± 0.003; all P<0.01). Furthermore, the rate of decrease was also much slower compared to the control group. In both control and experimental groups, IFN-γ and IL-10 secretion levels were observed to rise at first peaking at 24 hours and subsequently decrease. For each time period, IFN-γconcentration in the experimental group was significantly higher at 24 hours compared to the control group [(104.00 ± 1.07) versus (85.17 ± 0.28) pg/ml, P<0.01]. In contrast, IL-10 concentrations were significantly lower than the control group at all time points (P<0.01). IFN-γ/IL-10 ratio was also observed to be significantly higher than the control group at all measured time points (P<0.01). Conclusions (1) Recombinant protein LL-37 could significantly inhibit the growth of Candida. (2) By influencing the secretion of immune factors such as IFN-γ, IL-10, etc, recombinant protein LL-37 is able to adjust vaginal epithelial cells local immunity, and enhance resistance to Candida infection.

Objective To evaluate the role of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/nuclear factor E2-related factor 2 (Nrf2) signaling pathway in endotoxic shock-induced acute lung injury (ALI) in rabbits.Methods Thirty healthy male New Zealand white rabbits, aged 2 months, weighing 1.5-2.0 kg, were randomly divided into 3 groups (n =10 each) using a random number table: control group (group C), ALI group, and wortmannin group (group W).In group W, wortmannin 0.6 mg/kg (in 0.08 ml/kg dimethyl sulfoxide) was injected via the auricular vein, while the equal volume of normal saline was given in C and ALI groups.And 30 min later, lipopolysaccharide (LPS) 5 mg/kg (in 2 ml of normal saline) was injected via the auricular vein in ALI and W groups, while the equal volume of normal saline was given in group C.The rabbits were sacrificed at 6 h after LPS or normal saline injection.The lung was immediately removed for microscopic examination and for determination of wet/dry lung weight ratio (W/D ratio), expression of phosphorylated Akt (p-Akt) , Nrf2 and heme oxygenase-1 (HO-1) (by Western blot), and expression of Nrf2 and HO-1 mRNA (using fluorescent quantitative real-time reverse transcriptase-polymerase chain reaction).The pathological changes of the lung were scored.Results Compared with group C, the lung injury scores and W/D ratio were significantly increased, and the expression of p-Akt, Nrf2 protein and mRNA, and HO-1 protein and mRNA was up-regulated in ALI and W groups (P＜0.05).Compared with group ALI, the lung injury scores and W/D ratio were significantly increased,and the expression of p-Akt, Nrf2 protein and mRNA, and HO-1 protein and mRNA was up-regulated in group W (P＜0.05).Conclusion Activation of PI3K/Akt/Nrf2 pathway is the regulatory mechanism of the body adapting to the development of endotoxic shock-induced ALI in rabbits.

Objective To observe the effects of early rehabilitation on the motor function of post-stroke hemiplegic patients treated with intravenous thrombolysis.Methods Thirty-six post-stroke hemiplegic patients after intravenous thrombolysis with rtPA were recruited in this study.They were randomly divided into the early rehabilitation group (with the course less than three days) and the control group (with the course between three and seven days),18 cases in each group.Both group received a two-week rehabilitation programme.They were assessed using the Fugl-Meyer assessment (FMA) of up extremity,FMA of lower extremity and ADL score (modified Barthel index,MBI) before and after treatment,as well as at the 3rd month after treatment (during the following-up).Results After treatment and at the following-up,FMA scores of up extremity,FMA scores of lower extremity and MBI scores had improved significantly compared with before treatment in both groups.Furthermore,after treatment,in the early rehabilitation group the average FMA score of lower extremity and MBI score reached (23.33 ± 4.37) and (56.11 ± 22.27) respectively,significantly higher than those of the control group [(17.06 ± 4.70) and (40.00 ± 15.81) respectively].Conclusion Early rehabilitation contributes to short-term recovery of lower extremity motor function and ability in the activities of daily living in post-stroke hemiplegic patients after intravenous thrombolysis.