The epidermal growth factor receptor (EGFR) is overexpressed in many epithelial cancers,and is commonly caused by EGFR gene amplification and gene mutations. The most frequently occurring variant,the type III mutation (EGFRvIII) ,is characterized by an inframe deletion of exons 2-7 of the coding sequence. It is expressed only in tumors and not found in normal tissues, and therefore represents an attractive therapeutic target. The tumor therapy methods targeted for EGFRvIII include immu-notherapy, ribozyme, RNA interference, etc.

A fusion expression vector pPIC3.5K-PDGFR? was constructed to express recombinant receptor tyrosine kinase PDGFR? and the right Pichia pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of strain GS115, a high yield strain named M3 was screened. The strain M3 was cultured in a 5 L fermentor and His-GFP-PDGFR? fusion protein was purified by Ni2+ chelating affinity chromatography. One distinct peak was obtained after elution with 250 mmol/L imidazole. Fusion protein was proved to be 90.08 kD by western blotting, and have tyrosine kinase activity by ELISA. Results showed that the receptor tyrosine kinase PDGFR? was successfully expressed in P. pastoris and could be used as a target for small molecule selective inhibitors screening.

OBJECTIVE:To investigate the occurrence of ADR induced by Shuxuening injection in Beijing area,and to pro-vide reference for rational drug use in clinic. METHODS:ADR induced by Shuxuening injection in Beijing area during 2003-2013 was classified statistically in respects of population,organs/systems involved in ADR,clinical manifestations,occurrence time, drug combination and prognosis. RESULTS:Of 1648 ADR cases,the incidence of ADR in the elderly patients was higher than young patients,and that of female patients (60.92％) was higher than that of male patients (39.14％). Multiple organs/systems were involved in Shuxuening injection-induced ADR,and top 3 were nervous system (422 cases,21.33％),skin and its appen-dants (411 cases,20.78％) and others (305 cases,15.42％). Clinical manifestations were skin rash,itching,dizziness and ner-vousness,palpitation,arrhythmia,etc. ADR mostly occurred within 5-30 min after medication. In ADR reports,there were a vari-ety of drugs combined with Shuxuening injection. Shuxuening injection combined with Alprostadil injection was most common(61 cases,accounting for 23.46％ of drug combination). Among 1648 ADR cases,there were 622 cured cases and 1039 cases im-proved,4 cases were not improved,3 cases had sequelae and 1 case died. CONCLUSIONS:Clinical application of Shuxuening in-jection should strictly follow the indications of drug package inserts;differentiation of symptoms and signs is based on specific dos-age,solvent,route of administration,etc. It is necessary to strengthen the prescription check,and strengthen the observation after medication especially for the elderly and other special groups so as to reduce and avoid ADR.

AIM:To study the action of nucleoside diphosphate kinase A(NDPK-A)on the growth of S_ 180,H_ 22,Lewis and H460.METHODS:S_ 180 or H_ 22 cell(5?106)were inoculate subcutaneously into the right armpit of 85 Kunming mice,which were randomized into 8 groups.Lewis lung carcinoma cells(2?105)were inoculate subcutaneously into the right armpit of 85 C57BL/6 mice,which were randomized as Kunming mice.From the 2nd day,the treated groups were given different dose of rhNDPK-A once a day for 8 days(for S_ 180 or H_ 22 by iv)or for 10 days(for Lewis by ip),and the control group was given physiological saline only.H460 tissue pieces about 1.5 mm?1.5 mm?1.5 mm each were inoculated subcutaneously into the armpit of 38 Balb/c/neu mice.After the volume of xenograft become 100 mm?100 mm?100 mm,the nude mice were randomized into 5 groups and given different dose of rhNDPK-A once a day for 17 days.2 days after above treatments,the mice were killed and dissected.The knubs were peeled off and weighted.RESULTS:The growth of S_ 180,H_ 22 and H460 were inhibited by rhNDPK and the growth of H_ 22 was inhibited by rhNDPK at dose of 20 mg/kg combined with cisplatin(0.5 mg/kg).But the growth of Lewis lung cancer was not inhibited.CONCLUSION:rhNDPK-A inhibited the growth of S_ 180,H_ 22 and H460.rhNDPK-A(20 mg/kg)potentiated the antitumor action of cisplatin on H_ 22.

On the basis of the Uncaria transcriptome, specific primers were designed for UrSTR. The full-length cDNA of UrSTR (GeneBank: OL310251) was 1 541 bp, encoding 345 amino acid residues, and the promoter region sequence of UrSTR (GeneBank: OL310252) was 1 179 bp. Phylogenetic tree is revealed that UrSTR had a closest relationship with STR from Ophiorrhiza pumila and Ophiorrhiza japonica. Localization of UrSTR protein is revealed located in the vacuole membrane. Plant-care analysis indicated that the promoter region sequence of UrSTR, covering multiple light, stress and hormone-response cis-regulatory elements, and verified transcriptional activity. The results of SDS-PAGE show that pET-28a-UrSTR recombinant protein was successfully expressed, and the size was anticipated. The UrSTR prokaryotic expression system needs to be optimized in the later stage. The research lays the foundation for further purification to study its structure and functional characterization of the UrSTR protein.

OBJECTIVETo study the effect of acupuncture on the immune function of sepsis patients.

METHODSNinety sepsis patients were assigned to the control group, the thymosin a1 group, and the acupuncture treatment group according to random digit table, 30 patients in each group. Patients in the control group were treated according to the guideline of Surviving Sepsis Campaign (SSC). Patients in the control group received routine treatment. Those in the thymosin alpha1 group additionally received subdermal injection of thymosin alpha1 (1.6 mg), once per day for 6 successive days. Needling at related points such as Zusanli (ST36), Yanglingquan (GB34), Neiguan (PC6), Guanyuan (RN4), and so on, was performed in patients of the acupuncture treatment group, once per day for 6 successive days. T cell subgroups (CD3+, CD4+, CD8+, CD4+ /CD8+) and immunoglobulin levels (IgG, IgA, IgM) were detected. The length of ICU hospital stay, hospital readmission rate, and 28-day mortality were compared among the three groups.

RESULTSAfter six days of treatment, CD3+, CD4+, CD8+, IgG, IgA, IgM, and CD4+ /CD8+ ratio of three groups were all significantly increased (P < 0.01). Of them, CD3+, CD4+, CD8+, IgG, IgA, and IgM increased more significantly in the thymosin alpha1 group and the acupuncture treatment group (P < 0.01). Compared with the control group, the ICU hospitalization length was significantly shortened, the hospital readmission rate and the 28-day mortality were lower in the thymosin alpha1 group and the acupuncture treatment group (P < 0.05, P < 0.01). There was no statistical difference in each index between the thymosin alpha1 group and the acupuncture treatment group (P > 0.05).

CONCLUSIONAcupuncture could adjust the immune function of sepsis patients, improve their immunological indicators and prognoses.

Acupuncture Therapy ; CD4-CD8 Ratio ; Humans ; Length of Stay ; Prognosis ; Sepsis ; diagnosis ; immunology ; therapy ; T-Lymphocyte Subsets ; Thymosin ; analogs & derivatives

Objective To observe and analyze the MRI manifestations of Klippel-Trenaunay syndrome(KTS).Methods Thirty-one cases with diagnosed KTS underwent MRI on a 1.5 T MR system. MRI,MR venography(MRV),MR angiography(MRA)and X-ray venography(XRV)were performed.The pathological changes of the limbs and their veins were observed.Results MRI found soft tissue hemangiomas in 12 cases,soft tissue swelling of the extremities in 27 cases,superficial varicosities in 21 cases,and malformation of the veins in 27 cases.In twenty patients who underwent both MRV and XRV, superficial varicosities in 17 cases and persistent sciatic veins in 11 cases were found with both techniques. The increase of tributary veins was found in 10 cases with XRV,while found in 15 cases with MRV.The erratic venous course was found in 4 cases with MRV.The abnormalities of deep veins were found in 8 cases with MRV,while found in 7 cases with XRV.Conclusion MRI is an efficient and reliable imaging method for diagnosis of KTS.

Objective To isolate and identify the adult neural stem cells from the parenchyma of spinal cord in adult mouse.Methods The parenchymal spinal cord from adult mouse was dissected and dissociated by mechanical trituration.The tissue suspension was cultured in serum-free DMEM/F12 medium supplemented with EGF and B27.The cell colonies generated from a single cell were screened by limited dilution and incubated with BrdU.The cell colonies were transferred into medium with serum to induce differentiation.The cells were identified with antibodies to Nestin,BrdU,MAP2 and GFAP by immunofluorescence staining.Results The cells were cultured for seven days to generate proliferative neurospheres.The majority of cells in these neurospheres expressed Nestin and were differentiated into MAP2-positive cells and GFAP-positive cells in medium containing with fetal bovine serum.Conclusion A significant number of neural stem cells are present in the parenchymal adult mouse spinal cord and can proliferate and also give rise to neurons and glia in vitro.

OBJECTIVETo search for a simple and effective surgical approach to the management of moderate to severe pediatric concealed penis in children.

METHODSWe used Devine's technique via incision between the penis and scrotum in the treatment of 68 cases of moderate to severe pediatric concealed penis. The patients were aged 3 -13 (mean 6.5) years, 30 with moderate and 38 with severe pediatric concealed penis.

RESULTSThis strategy achieved good near- and long-term effects and satisfactory appearance of the penis, which was similar to that of circumcision. At 3 months after surgery, the penile length was 3 - 5.2 cm, averaging (2.35 +/- 0.35) cm.

CONCLUSIONDevine's technique via incision between the penis and scrotum is a simple and effective surgical option for moderate to severe pediatric concealed penis in children.

Adolescent ; Child ; Child, Preschool ; Humans ; Male ; Penis ; abnormalities ; surgery ; Scrotum ; surgery ; Urologic Surgical Procedures, Male ; methods

BACKGROUND:The effect of advanced glycation end products (AGEs) on bone resorption is controversial. Our previous study has shown that bone resorption is significantly inhibited when AGEs present with pre-osteoclast cells RAW 264.7, while the effect of AGEs on osteoclastic acidification remains unknown. OBJECTIVE:To investigate the effect of AGEs on osteoclastic acidification and the underlying mechanism. METHODS:RAW 264.7 cells were induced by RANKL (15μg/L;normal group) to generate osteoclasts, and AGEs (50-400 mg/L;experimental group) or bovine serum albumin (100 mg/L;control group) were added at the beginning of the induction. The effect of AGEs on bone resorption was evaIuated by anaIyzing the area of bone resorption on the Osteo Assay Surface plates, and the effect of AGEs on osteoclastic acidification was evaluated by acridine orange staining. Furthermore, the expression levels of V-ATPase a3 and CIC-7 were detected to investigate the underlying mechanism. RESULTS AND CONCLUSION:The bone resorption area in the AGEs group was significantly decreased compared with the normal group (P<0.05). Acridine orange staining reveaIed that the red fluorescence (620 nm) intensity in the AGEs group was significantly decreased compared with the normal group (P<0.05), and this inhibitory effect became obvious with the increase of AGEs concentration. Immunocytochemistry, western blot assay and PCR findings showed that the expression levels of V-ATPase a3 and CIC-7 in the AGEs group were decreased significantly compared with the normal group (P<0.05). To conclude, AGEs exert inhibitory effect on osteoclastic acidification, probably by inhibiting the expression of V-ATPase a3 and CIC-7.