Objective To research the mechanisms and the link of therapy and the optimal dosage of Tongru Sanpitang on hyperplasia of mammary glands in model rats. Methods Rats were randomly divided into 6 groups:negative contronl group, model group, Rupixiao Tablets group and Tongru Sanpitang low, middle, high dose group. Model was made with complex method of Progesterone and Estradiol benzoatehad injected into abdominal. In the middle of experiment, rats were given different drug respectively. Estradiol, follicle stimulating hormone, luteinizing hormone, the expression of estrogen receptor, weight of the ovary and uterus and visceral coefficient were determined. Results Follicle stimulating hormone, luteinizing hormone, the expression of estrogen receptor in medicine group decreased. The ovary and uterus in electron microscope, weight of organs and organ coefficient were improved. Conclusion Tongru Sanpitang may adjust reproductive endocrine function by decreasing sensitivity of mammary glands to estradiol and adjust whole function in the treatment of hyperplasia of mammary glands. There are dose-effect relationship.

Objective:To investigate job burnout status of doctor and nurse in China. Methods: 218 doctors and nurses were assessed by MBI-HSS. Results: 42.1% subjects got high or moderated scores on Emotional Exhaustion, 22.7% subjects got high or moderate scores on Depersonalization and 48.6% subjects got low or moderate scores on personal accomplishment. Female subjects got higher scores on Emotional Exhaustion than male subjects Compared with.subjects with higher level education, subjects with lower level education got higher scores on Emotional Exhaustion, however, lower scores on Personal Accomplishment.Conclusion:Some subjects are found to experiences job burnout. To prevent or cure job burnout, more attention should be paid to female and persons with lower level education.

Objective:To explore the relationship between work-family conflict and job burnout of doctors and nurses.Method:218 doctors and nurses were assessed by Work-Family Conflict Scale and Maslach Burnout Inventory.Result:Hierarchical regression indicated that the work-family conflict was a powerful predictor of job burnout beyond demographic variables, 37% variance of Emotional Exhaustion was explained by work-family conflict, 18% variance of Depersonalization and 12% variance of Personal Accomplishment were also explained by work family conflict. Conclusions:The work-family conflict is an important, may be the most influencing factor on job burnout in medical staffs.

In dietetic atherosclerotic models, contents of plasma atrial natriuretic peptide (ANP) and serum lipids were determined. The results showed that plasma ANP contents of the atherosclerotic group (14.33 ? 3.58?g/L)were higher than those of the control group (9.43 ? 3.14 ?g/L) (P

Objective:To construct double copy and x gene deleted HBV expression plasmid and study its expression in Hep3B cell line. Methods:The double copy HBV DNA ( adr Ⅰ) was used to inactivate HBV x gene by inserting mutation and gene recombination. The inserted 55 bp DNA sequence was synthesized artificially; the insertion point was ApaL Ⅰ of x gene area. After recombination, an x gene defected HBV plasmid containing single P, S and C gene was constructed,which can express in mammalian cell line. Another plasmid carrying double copy HBV DNA with normal x gene was constructed as contrast. Both were used to transfect Hep3B cells. Then the cells were screened by G418 and HBV virus in culture medium were isolated and detected by fluorescence quantitative PCR. Results: Plasmids pcDNA3 KN F1F2 and pcDNA3 ES HBV2 were constructed successfully. After cell transfection, the HBV DNA was highly expressed with both plasmids on the 3 rd, 6 th,14th day. Conclusion: The plasmids constructed can express in Hep3B cell line and cause HBV replication; x gene defected HBV gene has no effect on HBV replication in Hep3B cell line.

Objective　To study the relationship between infarct location and early neurological deterioration（END) in isolated subcortical infarctions.Methods　We collected patients who were admitted with a diagnosis of isolated subcortical infarction.According to their infarct location,patients were divided into proximal infarct group and distal infarct group.National Institutes of Health Stroke Scale (NIHSS) score was used to assess the degree of neurological impairment in both groups on the day of admission and within 3 days after admission.If the NIHSS score increased by 2 points or more within 72 hours after admission,END was considered.Baseline data and the incidence of END were compared between groups.Finally,We screened the independent risk factors for early prediction of END.Results　A total of 422 cases were collected,68 of 239 cases with proximal infarct and 13 of 183 cases with distal infarct developed END.There were statistically significant differences in age,male,LDL,history of stroke,proximal infarct and carrier artery stenosis in END,compared with non-END.Logistic regression analysis showed that age,male,history of stroke,proximal infarct and carrier artery stenosis were independent risk factors for END.Conclusion　Patients with proximal infarct are more likely to develop END in isolated subcortical infarctions,which should arouse clinical attention.

Objective To explore a promising system for tumor CD 44 receptor-targeted imaging and to investigate their physic-chemical properties and targeting effect on CD 44 abundant cancer cells in vitro.Methods The superparamagnetic iron oxide ( SPIO) nanoparticles were prepared by a coprecipitation in alkaline media starting from a mixed of the ferrous and ferric solution.And then the surface of the SPIO nanoparticles were modified with APTMS by a reaction with the hydroxyl groups.Finally, the hyaluronan-modified SPIO ( SPIO-HA) nanoparticles were prepared.Control and experimental groups were established after adding SPIO or SPIO-HA as agents respectively.Transmission electron microscopy ( TEM) and particle size analyzer were used to measure these nanoparticle sizes and the hydrodynamic diameters.Thermogravimetric analysis ( TGA) was carried out to evaluate the HA-content on the surface of SPIO-HA.The MRI T2 ralaxivities (1/T2 ) of the two groups at different Fe concentrations (0.09, 0.18, 0.27, 0.36, 0.45 mmol/L ) were measured on a 3.0T MR system.HepG2 cells and HL7702 cells were used for assessment of cells viability by methyl thiazolyl tetrazolium ( MTT ) assay.Prussian blue staining , immunoassay fluorescence image and flow cytometry were carried out to determine the targeted cellular uptake of SPIO-HA nanoparticles.MRI were performed to show the MR T 2 value changes after incubating with HepG2 cancer cells by using T 2 WI sequences at a clinical 3.0 T MR system.One-way analysis of variance was performed to determine significant changes in MR T 2 values of blank control , SPIO-HA and SPIO groups.Results The SPIO-HA and SPIO NPs were fairly homogeneous with an average core size of 18.2 and 22.4 nm, hydrodynamic diameter of 91.1 and 103.2 nm, Zeta potential of (-45.00 ±0.86) mV and (-18.50 ±0.73) mV, and magnetic relaxivity of 0.212 ×106 M-1 · s-1 and 0.191 ×106 M-1 · s-1.Based on the TGA data , HA accounted for 24%weight of each SPIO-HA.The internalization of the SPIO-HA was confirmed by prussian blue staining , while the cells showed no obvious blue stains with SPIO , incubation of SPIO-HA with tumor cells led to blue color inside the cells.After that, we examined cancer cell binding of FITC-SPIO-HA by immunoassay fluorescence image and flow cytometry.The green fluorescence resulting from FITC-SPIO-HA was observed inside the cells in both the cytoplasm and the plasmalemma.Tumor cells treated with SPIO-HA exhibited higher fluorescence signals with 7.97-fold enhancement observed for HepG 2 cells over control particles.In vitro MR, mean T2 values of blank control , SPIO and SPIO-HA groups were ( 115.20 ±0.36 ), ( 115.07 ±0.81 ) and ( 21.67 ±0.21 ) ms, respectively.There was significant difference among those three groups (F=31 703.339,P<0.01), MR T2 values of HepG2 cells treated with the SPIO-HA NPs were lower than blank and SPIO group.In comparison, SPIO did not generate any MRI signal changes compared with blank group.Conclusion The tumor CD44 receptor-targeted MR molecular probe SPIO-HA had a good physic-chemical property and well targeted HepG2 cells.

Objective To investigate the expression of Circulating tumor cells ( CTCs ) in peripheral blood cells of patients with different stages of Small-Cell Lung Cancer ( SCLC) ,and to evaluate its significance in early diagnosis of lung cancer metastasis.Methods Thirty-five patients with SCLC ( 12 in limited stage and 23 in extensive stage ) and 25 patients with benign lung disease were recruited at the Shanghai Changhai Hospital from April 2011 to April 2013.Samples were prepared from 7.5 ml peripheral venous blood and collected in CellSave tubes.The CTC counts were determined using CellTracks AutoPrep fluorescence scanning system according to the manufacturers′instructions.The expression of CTCs in peripheral blood of SCLC patients and benign lung disease patients were analyzed.The expression of CTCs in different stages of SCLC was measured and compared.Furthermore, samples were prepared from 2 ml peripheral venous blood by centrifugation.The serum levels of NSE Neuron specific enolase were measured.The relationship between the expression of CTC and NSE was analyzed.Results CTCs positive rates in SCLC were significantly higher than in benign lung disease controls [ Positive rates of CTC≥1 were 80.0%and 12.0%(χ2 =27.003,P<0.000 1),CTC≥5 were 51.4%and 0 (χ2 =18.367,P<0.000 1), CTC≥10 were 34.3%and 0(χ2 =10.714,P<0.001),CTC≥50 were 17.1 and 0(P=0.036,P<0.05), respectively ].CTCs positive rates in SCLC extensive stage were significantly higher than limited-stage [Positive rates of CTC≥1 were 58.3% and 91.3%(P=0.033,P<0.05), CTC≥5 were 65.2% and 25.0%(P=0.035,P<0.05), respectively].The expression of CTCs was significantly correlated with NSE (r=0.743 7, P<0.000 1).Conclusion The expression of CTC in SCLC patients is significantly higher than those in control group , and is closely related to clinical stages , which may provide new clues to early predicting of SCLC metastasis and deterioration.

OBJECTIVETo explore the genetic etiology of a patient with classic maple syrup urine disease (MSUD).

METHODSNext-generation sequencing (NGS) was used to screen the exons of BCKDHA, BCKDHB, DBT and DLD genes. Suspected mutations were verified by Sanger sequencing. Bioinformatic analysis was carried out to predict the influence of mutations on the protein structure and function.

RESULTSNGS and Sanger sequencing have detected a c.550delT mutation in exon 5 of the BCKDHB gene in the mother and a c.1046G>A mutation in exon 10 of the BCKDHB gene in the father, while no mutation was found with BCKDHA, DBT and DLD genes. Among these, the c.550delT is a novel mutation. Bioinformatic analysis suggested that the two mutations both located in a highly conserved region and may decrease the activity of branched-chain α-ketoacid dehydrogenase complex through alternation of its structure.

CONCLUSIONThe compound heterozygous mutations c.550delT and c.1046G>A of the BCKDHB gene probably underlie the clinical manifestations of the patient with classic MSUD.

OBJECTIVETo break immune tolerance to prion (PrP) proteins using DNA vaccines.

METHODSFour different human prion DNA vaccine candidates were constructed based on the pcDNA3.1 vector: PrP-WT expressing wild-type PrP, Ubiq-PrP expressing PrP fused to ubiquitin, PrP-LII expressing PrP fused to the lysosomal integral membrane protein type II lysosome-targeting signal, and PrP-ER expressing PrP locating the ER. Using a prime-boost strategy, three-doses of DNA vaccine were injected intramuscularly into Balb/c mice, followed by two doses of PrP protein. Two weeks after the last immunization, sera and spleens were collected and PrP-specific humoral and cellular immune responses evaluated by ELISA and ELISPOT tests.

RESULTSHigher levels of serum PrP antibodies were detected in mice vaccinated using the strategy of DNA priming followed by protein boosting. Of these, WT-PrP, Ubiq-PrP, and PrP-LII induced significantly higher humoral responses. ELISPOT tests showed markedly increased numbers of IFN-γ-secreting T cells in mice vaccinated using the strategy of DNA priming followed by protein boosting after stimulation with recombinant PrP23-90 and PrP23-231. PrP-ER induced the strongest T-cell response.

CONCLUSIONPrion vaccines can break tolerance to PrP proteins and induce PrP-specific humoral and cellular immune responses.

Animals ; Antibodies ; immunology ; CHO Cells ; COS Cells ; Cercopithecus aethiops ; Cricetinae ; Cricetulus ; Enzyme-Linked Immunosorbent Assay ; Female ; HeLa Cells ; Humans ; Immune Tolerance ; Interferon-gamma ; immunology ; Lysosomal-Associated Membrane Protein 2 ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Peptide Fragments ; immunology ; Prions ; genetics ; immunology ; Receptors, Peptide ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; immunology ; Recombinant Proteins ; immunology ; Transfection ; Ubiquitin ; genetics ; immunology ; Vaccines, DNA