Objective To prepare the immunotoxin protein (183B 2ScFvPE38) which might be useful in immuno guided therapy for ovarian carcinoma and study the activity of the protein. Methods The methods of ELISA and cytotoxicity were used to study the immunotoxin after induced with IPTG and the activity of the immunotoxin. Results The expressed fusion proteins were detected mostly as inclusion bodies at high level, and soluble immunotoxins were also observed. The results showed liable activity of antibody part and toxic part. Conclusion The recombinant fusion protein 183B 2ScFvPE38 keeps the activity of both components and might be of great use in the future to deal with ovarian carcinoma. [

Objective To study the relationship between soluble tumor necrosis factor receptor (STNFR) 1,2 and insulin resistance (IR). Methods STNFR1 and STNFR2 were measured by ELISA in 43 men and 41 premenopausal women. IR was assessed by Homa Model. Results Obese men and women showed higher levels of STNFR2 than nonobese men and women (P

Objective To evaluate the effect of BML?111 on ventilator?induced lung injury in rats. Methods Forty?eight healthy male Sprague?Dawley rats, weighing 200-250 g, aged 6-8 weeks, were randomized into 6 groups ( n=8 each) using a random number table: control group ( C group) , low tidal volume (VT) group (LVTgroup), high VT group (HVTgroup), low dose BML?111 group (BL group), high dose BML?111 group ( BH group) , and BML?111 plus BOC?2 ( lipoxin A4 receptor antagonist) group ( BOC?2 group) . Group C kept spontaneous breathing after tracheotomy, and received no mechanical venti?lation. The rats in the other 5 groups were mechanically ventilated ( respiratory rate 80 breaths∕min, frac? tion of inspired oxygen 21%, positive end?expiratory pressure 0) . The VT was 6 ml∕kg in group LVT , or 20 ml∕kg in HVT, BL, BH and BOC?2 groups. BML?111 0?1 and 1?0 mg∕kg were injected intraperitoneally during ventilation in BL and BH groups, respectively. In group BOC?2, BOC?2 50 μg∕kg was injected in?traperitoneally before ventilation, and BML?111 1?0 mg∕kg was injected intraperitoneally during ventilation. Arterial blood samples were collected at 4 h of ventilation, arterial oxygen partial pressure ( PaO2 ) was de?termined. Then animals were sacrificed by exsanguination. Bronchoalveolar lavage fluid ( BALF) of the left lung was collected for determination of neutrophil count, and the level of neutrophil was calculated. The right lung tissue specimens were obtained for microscopic examination, and for determination of wet∕dry lung weight ratio ( W∕D ratio ) , myeloperoxidase ( MPO ) activity, and contents of malondialdehyde ( MDA) , monocyte chemoattractant protein?1 ( MCP?1) , tumor necrosis factor?alpha ( TNF?α) , interleu?kin?1beta ( IL?1β) and IL?6. Results Compared with group C, PaO2 was significantly decreased, and the level of neutrophil in BALF, W∕D ratio, MPO activity, and contents of MDA, MCP?1, TNF?α, IL?1β and IL?6 were increased in group HVT ( P<0?05) , and no significant change was found in the variables mentioned above in group LVT ( P>0?05) . Compared with group HVT , PaO2 was significantly increased, and the level of neutrophil in BALF, W∕D ratio, MPO activity, and contents of MDA, MCP?1, TNF?α, IL?1β and IL?6 were decreased in group BH, and the contents of TNF?α, IL?1βand IL?6 were significantly decreased ( P<0?05) , and no significant change was found in the other variables in group BL ( P>0?05) . Compared with group BH, PaO2 was significantly decreased, and the level of neutrophil in BALF, W∕D ratio, MPO activity, and contents of MDA, MCP?1, TNF?α, IL?1β and IL?6 were increased in group BOC?2 (P<0?05). The pathological changes were significantly attenuated in group BL as compared with HVT and BOC?2 groups. Conclusion BML?111 can attenuate ventilator?induced lung injury in rats, and activated lipoxin A4 receptors are involved in the mechanism.

Objective To evaluate the effect of BML-111 on NF-κB pathway during acute lung injury induced by hemorrhagic shock and resuscitation in rats.Methods Thirty-two adult male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 4 groups (n =8 each) using a random number table:sham operation group (group S),hemorrhagic shock and resuscitation group (group HSR),BML-111 group,and BML-111 + BOC-2 (lipoxin A4 receptor antagonist) group (group BOC-2).The animals were anesthetized with intraperitoneal pentobarbital sodium.Hemorrhagic shock was induced by blood letting and maintained for 30 min.The animals were then resuscitated for 30 min by infusion of the shed blood and lactated Ringer's solution.In group BOC-2,BOC-2 (50 μg/kg) was injected intraperitoneally before blood letting.In BML-111 and BOC-2 groups,BML-111 (1 mg/kg) was injected intraperitoneally at the beginning of resuscitation.The rats were sacrificed at 2 h after the end of resuscitation and lungs were removed for determination of pathological changes,myeloperoxidase (MPO) activity,intercellular adhesion molecule-1 (ICAM-1) expression (by immunohistochemistry),tumor necrosis factor-alpha (TNF-α) content (by ELISA),and NF-κB p65 and IκB-α expression (by Western blot).Results Compared with group S,the MPO activity,ICAM-1 expression,and TNF-α content were significantly increased,NF-κB p65 expression was up-regulated,and IκB-α expression was down-regulated in group HSR.Compared with group.HSR,the MPO activity,ICAM-1 expression,and TNF-α content were significantly decreased,NF-κB p65 expression was down-regulated,IκB-α expression was up-regulated,and pathological changes of lung were attenuated in group BML-111.Compared with group BML-111,the MPO activity,ICAM-1 expression,and TNF-α content were significantly increased,NF-κB p65 expression was up-regulated,and lκ:B-α expression was down-regulated,and pathological changes of lung were aggravated in group BOC-2.Conclusion BML-1 11 inhibits activation of NF-κB pathway and inflammatory responses,thus mitigating acute lung injury induced by hemorrhagic shock and resuscitation in rats.

Objective To compare the consistency of thrombelastography (TEG) and light transmittance aggregometry (LTA) in monitoring the antiplatelet therapy of the patients with coronary heart disease (CHD) after percutaneous coronary intervention (PCI),and observe the changes of mean platelet volume (MPV) of the patients treated with aspirin and clopidogrel after PCI.Methods A total of 177 patients undergoing PCI and the treatment of aspirin and clopidogrel in Peking University First Hospital during March 2014 and May 2015 were enrolled in the study.Their adenosine diphosphate (ADP) or arachidonic acid (AA) induced platelet inhibition rates determined by TEG,MPV before and after antiplatelet therapy,and the maximum platelet aggregation rates measured by LTA from 99 patients were retrospectively analyzed.Results There was no any correlation between the maximum aggregation rates measured by LTA and the platelet inhibition rates determined by TEG regardless of using ADP or AA as agonist (all P ＞ 0.05).The detection rates of clopidogrel hyporesponsiveness determined by LTA and TEG were 30.3％ and 45.5％,respectively,while those of aspirin hyporesponsiveness were 19.2％ and 31.3％,respectively.The detection rate of hyporesponsiveness determined by LTA was significant lower than that by TEG (P ＜ 0.05).The MPVs after antiplatelet therapy were significant lower than that before treatment (all P ＜ 0.01) regardless of clopidogrel hyporesponsive or sensitive and aspirin hyporesponsive or sensitive.The MPVs in clopidogrel hyporesponsive group before and after treatment were significantly lower than that in clopidogrel sensitive group (all P ＜ 0.05).The PLT counts in clopidogrel or aspirin hyporesponsive groups after treatment were significantly higher than that before treatment (all P ＜ 0.05).Conclusion There is poor correlation between LTA and TEG.It should be noted that the incidence rate of antiplatelet drug hyporesponsiveness is high in clinical practice.The MPVs of the patients significantly decrease after antiplatelet therapy.The patients with a significant increase of PLT after antiplatelet therapy are more likely to become drug hyporesponsiveness,while the patients with lower MPV are more likely to have clopidogrel hyporesponsiveness.

Objective To evaluate the effect of lipoxin A4 receptor agonist BML-111 on acute lung injury induced by hemorrhagic shock and resuscitation in rats.Methods Thirty-two healthy male Sprague-Dawley rats,aged 6-8 weeks,weighing 200-250 g,were randomized into 4 groups (n =8 each) using a random number table:sham operation group (S group),hemorrhagic shock/resuscitation group (HSR group),BML-111 group,and BML-111 plus BOC-2 (lipoxin A4 receptor antagonist) group (BOC-2 group).The animals were anesthetized with 2％ pentobarbital sodium 80 mg/kg,tracheostomized and mechanically ventilated.Left common carotid artery was cannulated for blood-letting and fluid infusion.Hemorrhagic shock was induced according to the method described by Kochanek et al.MAP was reduced to 35-45 mmHg and maintained at this level for 30 min.The animals were then resuscitated for 30 min with infusion of the blood withdrawn and lactated Ringer' s solution 2 times the volume of blood withdrawn.In BML-111 and BOC-2 groups,BML-111 (1 mg/kg) was injected intraperitoneally at the beginning of resuscitation.In BOC-2 group,BOC-2 (50 μg/kg) was injected intraperitoneally before blood-letting.The rats were sacrificed at 2 h after completion of resuscitation.Bronchoalveolar lavage fluid (BALF) was collected for determination of neutrophil count.Lungs were excised for microscopic examination of the pathological changes and for determination of wet/dry lung weight ratio (W/D ratio),contents of interleukin-1 (IL-1β) and IL-6,and phosphorylation of mitogen-activated protein kinase (MAPK).Results Compared with group S,the neutrophil count in BALF,W/D ratio,contents of IL-1β and IL-6,and phosphorylation of MAPK were significantly increased in HSR group (P ＜ 0.05).The neutrophil count in BALF,W/D ratio,contents of IL-1β and IL-6,and phosphorylation of MAPK were significantly lower in BML-111 group than in HSR group,and higher in BOC-2 group than in BML-111 group (P ＜ 0.05).Conclusion BML-111 can attenuate acute lung injury induced by hemorrhagic shock and resuscitation in rats and inhibition of activation of MAPK pathways and reduction of inflammatory responses in lung tissues are involved in the mechanism.

The impact of acupuncture for the pregnancy of in vitro fertilization-embryo transfer (IVF-ET) is discussed in the paper. Nowadays there are various conclusions about the impact of acupuncture for IVF-ET, and it may result from the differences in research designs. The effect is closely related to the demographic and clinical characteristics of subjects, such as age, the diagnosis of barrenness, blood flow index of uterine spiral arteries, the cycle of IVF, etc. Besides, the efficacy is influenced by treatment based on syndrome differentiation or not, the frequency and course of acupuncture in both the treating group and the control group, etc. If more reasonable design is achieved in the further study based on them, more reliable evidence will be provided for the effect and mechanism of the pregnancy of IVF-EF by acupuncture.
Acupuncture Therapy ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; physiopathology ; therapy ; Male ; Pregnancy ; Pregnancy Rate ; Randomized Controlled Trials as Topic ; Treatment Outcome

Angiotensin II ; immunology ; Chronic Disease ; Heart Failure ; drug therapy ; Humans ; Renin-Angiotensin System ; immunology ; Vaccines ; therapeutic use

OBJECTIVETo investigate the status of Notch signaling pathway in small cell lung cancer (SCLC).

METHODSExpression plasmids of pEFBOS-NIC-MYC and pEFBOS-neo were transfected into NCI-H446 cells. Stably transfected cell lines were selected and their growth rates were examined by MTT method. Expression of downstream genes along the Notch signaling pathway were studied by RT-PCR. Protein expression of euroendocrine markers of CgA and NSE were detected by Western blot analysis and immunocytochemistry.

RESULTSThe expression of HES1 was increased in the pEFBOS-NIC-MYC group, but the expression of hASH in the pEFBOS-NIC-MYC group was decreased significantly. The transfected cells with pEFBOS-NIC-MYC plasmid showed a significantly slower growth rate compared with that of two control groups (P < 0.05, Student's t-test). Immunocytochemistry of NSE showed that PUs in the NIC transfected group, sham group and negative control group were 7.21 ± 0.59, 28.25 ± 1.46, 30.57 ± 1.31 respectively, the former one was smaller than the values of the latter two significantly (P < 0.01). Western blot analysis showed the grave scales of CgA in NIC transfected group and sham group to be 0.54 ± 0.03 and 0.99 ± 0.05 respectively (grave scales of the negative control was set as 1.00), the former one significantly smaller than that of the other two groups (P < 0.01). The grave scales of NSE in the NIC transfected group and sham group were 0.43 ± 0.02 and 1.07 ± 0.09 respectively (grave scales of the negative control was set as 1.00) and the former one was significantly smaller than the other two groups (P < 0.01).

CONCLUSIONNotch signaling pathway regulates SCLC cells through its inhibitory effect on hASH1 transcription via HES1 along with an expression inhibition of neuroendocrine markers in SCLC.

Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Chromogranin A ; metabolism ; Homeodomain Proteins ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Plasmids ; Receptor, Notch1 ; metabolism ; physiology ; Recombinant Proteins ; metabolism ; Signal Transduction ; Small Cell Lung Carcinoma ; metabolism ; pathology ; Transcription Factor HES-1 ; Transfection

OBJECTIVETo investigate the effects of Notch1 signal activation on proliferation and neuroendocrine marker expression in small cell lung cancer cells.

METHODSThe active form of Notch1 (NIC) was over-expressed in NCI-H446 cells by constitutive transfection and a stable transfected cell line was established. Proliferation of NCI-H446 cells was analysed by MTT assay on 6 successive days. Expression of neuroendocrine markers (CgA, NSE) was observed by immunohistochemistry and Western blot analysis. Statistical analysis was conducted to compare the results in cells with NIC transfected and those in control groups.

RESULTSMTT assay showed that absorbance (A) of cells overexpressing Notch1 was significantly depressed compared with that of the control cells (P<0.05). Immunohistochemistry of CgA showed that PUs in the NIC transfected group, sham group and negative control group were 8.81 +/- 0.77, 38.10 +/- 1.55, 38.97 +/- 0.80, respectively, the former one was significantly smaller than that of the latter two (P<0.01). Immunohistochemistry of NSE showed that PUs in the NIC transfected group, sham group and negative control group were 7.21 +/- 0.59, 28.25 +/- 1.46, 30.57 +/- 1.31, respectively, the former one was significantly smaller than that in the latter two (P<0.01). Western blot analysis showed that the gray scales of CgA in the NIC transfected group and sham group were 0.54 +/- 0.03 and 0.99 +/- 0.05, respectively, (gray scale of the negative control set as 1.00), the former one was significantly smaller than that of the other two groups (P<0.01). The gray scales of NSE in the NIC transfected group and sham group were 0.43 +/- 0.02 and 1.07 +/- 0.09, respectively (gray scale of the negative control set as 1.00), the former one was significantly smaller than that of the other two groups (P<0.01).

CONCLUSIONNotch1 may behave as a tumor suppressor in small cell lung cancer. Notch1 signal activation can inhibit the proliferation and neuroendocrine marker expression in small cell lung cancer cells, suggesting that Notch1 gene could be a new target for small cell lung cancer treatment and probable relief of paraneoplastic syndrome.

Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Chromogranin A ; metabolism ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Receptor, Notch1 ; genetics ; metabolism ; physiology ; Recombinant Proteins ; genetics ; metabolism ; Small Cell Lung Carcinoma ; metabolism ; pathology ; Transfection