Objective To construct recombinant lentivirus silence vector aiming at rictor gene in mTORC2 specific protein, and to investigate its regulation on mTORC2/SGK1 signal pathway and the effect on pulmonary alveolar epithelial sodium ion chan-nel,as well as the role in acute respiratory distress syndrome(ARDS)and acute lung injury.Methods The interfering vector plas-mid and empty vector plasmid of target gene rictor were constructed,which and the lentivirus packaging system were co-transfected to 293T cells.The viral supernatant was collected,centrifuged,concentrated and purified for obtaining recombinant lentivirus.The virus titer was detected and the virus was infected to A549 cells.Stable cell lines were screened.RT-PCR was used to confirm the silencing situation of target gene rictor.The expression situation of various signal indexes in this pathway was detected by PCR and Western blot.Results The recombinant lentivirus of silence gene rictor was successfully constructed and transfected to A549 cell for obtaining stable cell lines.Compared with blank and control groups,the mRNA levels of rictor,downstream SGK1 andα-,β-andγ-ENaC in the shRNA-rictor group were significantly decreased (P <0.05 ).Meanwhile,the protein levels of rictor,downstream SGK1,P-SGK andα-,β-andγ-ENaC in the shRNA-rictor group were significantly decreased compared with the other two groups(P<0.05).Conclusion Silence rictor gene has the obvious regulation effect on mTORC2/SGK1 signal pathway,meanwhile affects the expression of pulmonary alveolar epithelial cellular α-,β-and γ-ENaC at gene and protein level.It is speculated that mTORC2/SGK1 may be an important signal pathway for regulating the clearance capacity of pulmonary alveolar epithelial cells on pulmonary alveolar fluid and simultaneously affecting the pulmonary edema formation.

Objective To gain the virus like particles (VLPs) based on Trop2 targets ,which provided the basis for further in vi‐vo induced experiments and anti‐tumor vaccine studies .Methods Using molecular cloning method to constract eukaryotic expres‐sion vector of pCAGGs/Trop2 and baculovirus expression vector of pFastbac1/Trop2 ,expression product of pCAGGs/Trop2 in HeLa cells were intended to be to be anchored to the cell membrane by the methods of Immunohistochemistry ;pFastbac1/Trop2 were transformed into E .coliDH10bac isolates to gain recombinant bacmids ,which were transfected into insect cells to express re‐combinant baculovirus with Gag rBV ,then sucrose density gradient centrifugation ,Western Blot and electron microscope were per‐formed to purify and identify the Trop2 VLPs .Results Building recombinant bacmids which were transfected into insect cells to express recombinant baculovirus with Gag rBV ,gained the recombinant Virus like Particles .Conclusion Trop2 VLPs was success‐fully prepared ,which laid the foundation for the subsequent induction of humoral and cellular immune response .

The expressions of estrogen receptor (ER) α,ERβ1,and ERβ2 in thyroid carcinoma,thyroid adenoma,and adjacent normal thyroid tissue were determined by RT-PCR and immunohistochemistry,and the relationship of expression levels of three ERs and the main clinical pathologic parameters were evaluated.The result showed that ERα mRNA expression in papillary thyroid carcinoma(PTC) tissues was significantly higher than adjacent normal tissue,while,ERβ2 mRNA expression in PTC tissues was significantly lower than normal tissue ( P＜0.05 ).The positive expression of ERβ1 and ERβ2 in the thyroid carcinoma was related with the histological variant of carcinomas; the ERβ2 expressions were related with the lymph node metastasis and TNM staging of the carcinomas.These results suggest that the decreased ERβ2 expression may induce proliferation and carcinogenesis of thyroid follicular epithelial cells as well as progression of the carcinoma,and is a powerful prognostic indicator.

Objective To compare the dosimetric parameters between the use of Tandem and Ring (TR;Nucletron#090.617) or Tandem and Ovoid (TO;Nucletron#189.730) applicators during three-dimensional (3D) high-dose rate (HDR) brachytherapy (BT) for cervical cancer.Methods The records of 40 cervical cancer (ⅡB-ⅣA) patients treated with 3D-image-guided HDR-BT were reviewed.Of these 40 patients, 20 were treated with the TO applicator, and 20 with the TR applicator.The D100% and V150% of the clinical target volume (CTV) and the D2 cc of organs at risk (OAR)(the rectum, bladder, and small intestine) during 3D-HDR-BT using TO and TR were compared using the independent sample t-test.ResultsOverall metrics:CTV volume:66.04±13.86 cm3(TR) vs.65.67±15.08 cm3(TO)(P=0.052);CTV D100:3.71±0.34 Gy (TR) vs.3.37±0.49 Gy (TO)(P=0.016);CTV V150%:0.54±0.02(TR) vs.0.56±0.04(TO)(P=0.034);rectum D2 cc:3.38±0.30 Gy (TR) vs.2.95±0.80 Gy (TO)P=0.037);bladder D2 cc:4.33±0.39 Gy (TR) vs.2.93±1.27 Gy (TO)(P=0.00);and small ntestine D2 cc:3.04±1.02 Gy (TR) vs.3.41±0.57 Gy (TO)(P=0.171).Conclusions TR has better CTV coverage than TO during 3D HDR brachytherapy for cervical cancer.In addition, D2 cc of the rectum and bladder were both igher with TR than with TO, though there is no significant dosimetric difference in the small intestine between the two applicators.Therefore, tumor location, extent of invasion, and vaginal conditions should be considered when selecting the suitable pplicator for the treatment of cervical cancer.

Objective Delayed xenograft rejection (DXR) is a major barrier to the long-term xenograft survial.This study evaluated the interaction between human peripheral blood mononuelear cells (PBMC) and porcine endothelial cells (PEC),and the effects of new generation of rabbit antihuman leukocyte polyclonal antibody (newRALG) inhibiting xenogeneic cell-mediated immune responses.Methods newRALG was obtained from rabbits after immunization with activated lymphocytes and monoeytes.PEC were isolated from aorta,and human PBMC were isolated from peripheral blood.Co-cultures of PKH-26 labeled PEC with PBMC were established,newRALG,thymoglobulin,isotype Ig and scavenger receptor (SR) ligand poly G were added into the co-cultures.Cells were collected,then FACS analysis was carried out to detect the up-take of PEC membrane by monocytes and the expression of costimulatory molecules.Lymphocyte proliferative responses to PEC with or without antibody were evaluated by a xenogeneie mixed lymphocyte-endothelial cell reaction (xMLER).Results FACS analysis revealed that monocytes from PBMC-PEC co-cultures became positive for PKH-26 following their interaction with PKH-26 labeled PEC,indicating that they engulfed PEC membranes during activation.PKH-26 positive monocytes up-regulated the CD40 and CD80 expression.Furthermore,SR blockade with poly-G prevented PEC membrane up-take by monocytes,newRALG greatly reduced SR-mediated PEC membrane up-take.The effects of thymoglobulin in inhibiting PEC membrane uptake were limited.xMLER demonstrated strong lymphocyte proliferation in response to PEC,and lymphocyte proliferation was dramatically inhibited by newRALG but not isotype Ig at a dosmdependent manner.Conclusions Monocytes play an important role in xenogeneic immune responses.SR ligand poly G inhibits PEC membrane up-take.newRALG inhibits PEC membrane up-take by monocytes,suggesting that newRALG blocks SR.Additionally,newRALG inhibits lymphocyte proliferation in response to PEC.These results suggest that this new polyclonal preparation may thus impair the initiation of xeno-specific immune responses and prevent xenograft rejection.

Objective To investigate the dosimetric influence of dwell weight standard deviation (DWSD) and applicator displacement in cervical cancer patients treated with three-dimensional brachytherapy.Methods A total of 20 cervical cancer patients who had completed radical treatment were selected in this study.The Fletcher applicator (Nucletron#189.730) was used for these patients.A new plan,based on the former CT images and structures,was designed for each patient.In former and new plans,dwell weight was recorded,and DWSD was calculated.Two groups,low-DWSD (LDWSD,0.141-0.299) and high-DWSD (HDWSD,0.211-0.337),were set according to the DWSD size for the two plans.Dosimetric effects from ± 1 mm displacement of tandem applicator or ovoid applicator were simulated with Oncentra (R) Brachy V4.3 treatment planning system.D100,D90,and V150 for clinical target volume (CTV)and D0.1cc,D1cc,and D2cc for the bladder,rectum,and sigmoid were evaluated.Dosimetric comparisons were made between the LDWSD group and HDWSD group to study the dosimetric effects of DWSD and applicator displacement in cervical cancer patients.Results The dosimetric effects from applicator displacement increased with increasing DWSD.If there was a 1 mm displacement of tandem applicator or ovoid applicator,D100,D90,and V150 of CTV were 3.0％,23.8％,and 4.8％ higher or 0.5％,1.2％,and 5.2％ higher in the HDWSD group than in the LDWSD group;D0.1cc,D1cc,and D2cc of the bladder and rectum were significantly higher in the HDWSD group than in the LDWSD group,particularly for the sigmoid (up 44.0％,22.8％,and 16.8％) and (up 10.3％,14.4％,and 12.4％).Conclusions DWSD should be considered in plan evaluation for cervical cancer patients treated with three-dimensional brachytherapy.The dosimetric influence from applicator displacement can be decreased by reducing DWSD properly.

Adult ; Female ; Hearing Loss, Sensorineural ; complications ; Humans ; Mitochondrial Encephalomyopathies ; complications

Adolescent ; Catheter Ablation ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Tachycardia, Supraventricular ; surgery

Objective To observe the lung protection of Astragalus membranaceus against radiotherapy to intermediate-stage and terminal thoracic neoplasm, and its influence on TNF-α and ET expression.Methods The patients with intermediate-stage and terminal thoracic neoplasm under radiotherapy were divided into a treatment group and a control group.Patients in the treatment group took 10 ml of Asragalus membranaceus twice a day.for consecutive 6 months from the beginning of radio therapy.TNF-α and ET in the plasma were measured before and after the radiotherapy.The clinical symptom,iconographic changes and lung diffusion were observed from the 15th day of radiotherapy.Results The TNF-α and ET in plasma afterthe radiotherapy were(2.48±0.75)as/ml and(69.32±23.03)pg/ml for the treatment group,and(5.12±1.01)ns/ml and(97.87±37.83)pg/ml for the control group with the statistial difference(x2=7.49,6.57,P<0.001).The decrease of CO diffusion 5 and 10 months after the radiotherapy in the treatment group was statistically different compared with that in the control group(x2=3.98,3.78,P<0.05).There was a statistical difference of the incidence of acute radiation pneumonitis and pulmonary fibrosis between these two groups(P<0.05).Conclusions Astragalus membranaceus could inhibit the excess expression of TNF-α and ET in plasma and reduce the deterioration of diffusion after radiotherapy,so that it can be used for intervention of lung injuries from radiotherapy.