Objective:To evaluate the prognostic value of post-mastectomy radiation therapy (PMRT) in patients with axillary lymph node-positive breast cancer treated with breast conservation surgery and to establish the candidates for PMRT based on different pN stages and lymph node ratios (LNR). Methods:A retrospective analysis of the clinical data of the patients was conducted. The pa-tients had positive lymph nodes (n=152) between 1998 and 2007 and underwent breast conservation surgery. A comparison of the dis-ease-free survival (DFS) and overall survival (OS) rates was conducted based on LNR and pN staging and with PMRT as a prognostic factor. Results:A total of 152 cases were studied, of which 114 were pN1, 23 were pN2, and 15 were pN3. Among these cases, 114 had an LNR ranging from 0.01 to 0.20, 26 had an LNR from 0.21 to 0.65, and 12 had an LNR>0.65. Univariate analysis showed that the number of dissected lymph nodes, LNR, pN stage, estrogen and progesterone receptor status, and radiotherapy were the prognostic fac-tors for DFS and OS rates (P<0.05). Age and chemotherapy were prognostic factors only for OS rate (P<0.05). Multivariate analysis in-dicated that PMRT and LNR were independent prognostic factors of DFS and OS (P<0.05). The pN staging had no significant effect on DFS or OS (P>0.05). In the subgroup analysis, PMRT had significant effect on DFS and OS (P<0.05) in patients with pN1 and LNR<0.21. Conclusion:LNR is an independent prognostic factor in axillary lymph node-positive breast cancer patients treated with breast conservation surgery, and a candidate for PMRT should be established based on different LNR risks.

Axillary reverse mapping (ARM) is a technique used to map and preserve arm lymphatic drainage during axillary lymph node dissection (ALND) and/or sentinel lymph node dissection (SLND). As a result, the risk of arm lymphedema is reduced. ARM is an emerging technology for breast cancer surgery and currently in the clinical trial phase. In this article, related clinical trials conducted in recent years were reviewed and the displaying methods of ARM lymph nodes and lymphatic vessels were analyzed. The feasibility of retained ARM lymph nodes and lymphatic vessels was also evaluated. Furthermore, the clinical significance of ARM was evaluated in terms of the reduction of the incidence of upper extremity lymphedema after breast cancer surgery.

The overexpression of proto-oncogene protein c-pim-1 (Pim-1) in tumor tissue is related to the stage and prognosis.Recent studies indicate that Pim-1 plays a critical role in the proliferation and apoptosis of cells and the metastasis of tumor.Pim-1 acts as an essential factor in several signaling pathways and its expression and activation are regulated by many factors as well as affects others widely.As an influential factor in the occurrence and development of tumor,Pim-1 has been a potential target in oncotherapy.

ObjectiveTo investigate the possible pathogenesis of EB virus (EBV) latent membrane protein 1 in inducing systemic lupus erythematosus (SLE).MethodsThe mRNA expression levels of LMP1 and apoptosis-related genes bcl-2,bax in SLE patients and healthy controls were detected by real-time fluorescence quantitative polymerase chain reaction (PCR).The serum BAFF levels of SLE patients and normal healthy controls were detected by ELISA.2 test was used for positive rate analysis,2-△△Ct method was used for comparing the gene expression level,and Student-Newman-Kqeuls method was used for pair-wise comparison between the means.Results① The positive rate of LMP1 expression in 67 SLE cases was 25％,which was significantly higher than the 11％ in 65 healthy controls (P＜0.05).② The 2-△Ct value of bcl-2 mRNA expression level of SLE patients was 0.0257,1.41 times to that (0.0183) of healthy controls and the difference was statistically significant.③ The 2-△Ct value of bcl-2 mRNA expression level of LMP1 positive SLE patients was 0.0427,1.98 times to that of LMP1 negative SLE patients (0.0217),the difference was statistically significant.④ The serum BAFF levels of LMP1 positive SLE patients,LMP1 negative SLE patients,LMP1 positive healthy controls and LMP 1 negative healthy controls were ( 106± 15 ),(82± 19),( 68±19),(64±17) μg/L,respectively.There were significant differences between serum BAFF levels of LMPl-positive SLE patients and other groups(P＜0.0l ).There were significant difference between serum BAFF levels of LMP1-negative SLE patients and the control groups (P＜0.01).ConclusionEBV may induce and/or promote SLE by LMP1 through apoptosis-related genes bcl-2 expression and induction of B lymphocytes that produce BAFF,all these mechanisms can prolong the infected auto-reactive B lymphocytes survival.

Objective:To construct recombinant lentiviral vectors harboring interference RNA ( RNAi ) targetting murine TNF-αgene,so as to lay the foundation on the RNAi gene therapy.Methods: Three small interfering RNA ( siRNA) sequences targeting murine TNF-αgene ( siRNA1,siRNA2,siRNA3) and negative-control siRNA were designed and synthesized.The inhibition effects of siRNAs on TNF-α,IL-1βand IL-6 secretion of LPS-stimulated RAW264.7 macrophages were observed using real-time PCR and ELISA methods.DNA oligo was designed and synthesized according to the most effective siRNA 2 sequence.The recombinant lentiviral shuttle plasmid expressing short hairpin RNA ( shRNA) was constructed and sequenced.The lentiviral shuttle plasmids with packaging plasmids were transfected into 293T cells to produce lentiviral particles.Results: ①The TNF-αmRNA relative expression levels of siRNA1, siRNA2 and siRNA3 were 0.24±0.01,0.16±0.02,0.19±0.01 respectively,significantly lower than that of negative control (0.95± 0.02) (F=531.3,P<0.001).The inhibition rates at mRNA level were 74.26%,83.09%,79.93%,respectively comparing with negative control.No significance was observed in IL-1βor IL-6 mRNA relative expression change after TNF-αsiRNA transfection ( P>0.05).②The TNF-αprotein expression levels of siRNA1,siRNA2 and siRNA3 were (23.95±1.21),(17.27±1.46),(19.07± 1.57)ng/ml respectively,significantly lower than that of negative control (35.37±2.93)ng/ml (F=18.1,P=0.000 6<0.001).The inhibition rates of protein expression were 32.29%, 51.16%, 46.08%, respectively comparing with negative control.③The PCR product electrophoresis showed that recombinant vectors yielded 343 bp fragments,non-constructed vectors yielded 306 bp fragments.DNA sequencing partially showed insertion sequence.④Lentiviral particles were obtained by transfecting 293T cells with recombinant lentiviral shuttle plasmids and lentiviral packaging plasmids.Cells grew well during virus production with strong fluorescence expression.The titer of concentrated virus was 2×106 TU/μl.Conclusion:The lentiviral vector harboring RNAi targeting murine TNF-αgene has been successfully constructed.

Objective:To retrospectively evaluate the prognostic risk factors of T1-2 stage breast cancer patients with one to three positive node(s) and their effects on the benefits of post-mastectomy radiation therapy (PMRT). Methods:We retrospectively analyzed 457 breast cancer patients with T1-2 stage and one to three positive axillary lymph nodes treated in our hospital between 2000 and 2002. The independent prognostic factors of the patients were calculated by the Cox proportional hazards model. The patients were fur-ther classified into high-risk and low-risk subgroups according to the risk factors to explore the benefit of PMRT on the prognosis of dif-ferent subgroups using survival analysis. Results:PMRT was not an independent beneficial factor of overall survival (OS) (HR=0.949;CI:0.435-2.074;P=0.896) or loco-regional recurrent free survival (LRRFS) (HR=0.611;CI:0.231-1.614;P=0.320) in all patients. Ex-tracapsular extension (ECE) and pathological grades were independent prognostic risk factors, and the benefits of PMRT were signifi-cantly different on the prognosis of high-risk subgroup patients (group ECE+OS:P=0.020, LRRFS:P=0.014;group GradeⅢOS:P=0.002, LRRFS:P<0.001). Meanwhile, PMRT failed to prolong the OS and LRRFS of low-risk subgroup patients (group ECE+OS:P=0.353, LRRFS:P=0.796;group GradeⅠtoⅡOS:P=0.267, LRRFS:P=0.589). Conclusion:ECE and gradeⅢwere the independent risk factors of death and loco-regional recurrence in the T1-2 breast cancer patients with one to three positive lymph node(s). PMRT was an effective adjuvant therapy to improve the prognosis of patients with high-risk factors. However, the benefit of PMRT had no sig-nificance in patients with ECE-or gradeⅠ-Ⅱ.

Objective:To synthesize a new β-amyloid (Aβ) radioactive tracer (2-((2-6-[ 18F]fluoro-5-(methylamino)pyridin-2-yl)benzothiazol-6-yl)thio)ethanol ( 18F-FINH-Me), and evaluate its biological distribution and affinity to Aβ plaques. Methods:18F-FINH-Me was synthesized by GE FN automated module, and the quality control and stability of 18F-FINH-Me were determined with high performance liquid chromatography (HPLC). The biodistribution of 18F-FINH-Me was observed in normal C57BL/6 mice ( n=25). MicroPET/CT imaging was performed in Alzheimer′s disease (AD) model mice( n=5) and matched normal C57BL/6 mice( n=5). The brain tissues of mice were taken for Aβ immunohistochemical staining. 18F-FINH-Me autoradiography was performed in postmortem brain sections of one AD patient (female, 69 years old) and one healthy volunteer (female, 66 years old). Results:The decay correction yield of 18F-FINH-Me was (53±4)% ( n>20) with the radioactive purity of more than 98% ( n>20) and the specific activity of 79.90-122.00 GBq/μmol ( n=10). 18F-FINH-Me was stable in phosphate buffered solution (PBS) after incubation for 4 h at room temperature. The biodistribution showed that 18F-FINH-Me was mainly excreted through the liver and kidneys. MicroPET/CT imaging showed that 18F-FINH-Me was obviously uptaken in the brain of AD mice. After injection for 1-2 min, the uptake of 18F-FINH-Me reached the peak, and the elution speed was fast (whole brain standardized uptake value: 0.73±0.17 for 1 min, 0.31±0.06 for 30 min). The immunohistochemistry showed that there were abundant Aβ plaques in the brain of AD model mice but not in the normal C57BL/6 mice brain. The autoradiographic results showed that 18F-FINH-Me exhibited substantial plaque labeling in brain sections of one AD patient but not in the healthy volunteer. Conclusion:18F-FINH-Me may be an effective PET agent for detecting Aβ plaques in brain.

OBJECTIVETo investigate the flavonoids in Rhododendron mariae.

METHODThe constituents in R. mariae were determined by UPLC/Q-TOF-MS. The chromatographic separation was performed on a C18 column (2.1 mm x 50 mm, 1.7 microm) with a gradient elufion of methanol-water containing 0.1% formic acid. The mass specfrometer eqaipped with elecfrospay ionizafion source was usedas defecfor and operated in data was collected under the positive and negative ion modes.

RESULTSeven constituents were identified as myricetin-3-O-beta-D-gluconside (1), myricetin-3'-O-beta-D-xylopyraoside (2), hyperoside (3), isoquercitrin (4), avicularin (5), quercitroside (6) and quercetin (7).

CONCLUSIONIn this study, the main flavonoids in R. mariae were separated by UPLC, and identified through the information of positive ion and negative ion and relative molecular mass which were determined by Q-TOF-MS. It is an accurate and effective method which can be applied for the constituent identification of R. mariae.

Chromatography, High Pressure Liquid ; Flavonoids ; analysis ; chemistry ; Mass Spectrometry ; Rhododendron ; chemistry

OBJECTIVETo establish HEK293 cell lines with stable expression of human parathyroid hormone (PTH) receptors.

METHODSThe purified gene fragments of PTH-related peptide receptor (PTHR) and its mutant form (DSEL) were cloned separately into pcDNA3.1(+) vector after digestion with EcoR I and Not I, and the resulted pcDNA3.1(+)-PTHR and pcDNA3.1(+)-DSEL plasmids were verified by restriction enzyme digestion and DNA sequencing. HEK293 cells were transfected with these plasmids and the expression of PTHR and DSEL in the cells were examined by RT-PCR and ELSIA.

RESULTSSequencing and restriction enzyme digestion analysis showed that PTHR and DSEL cDNAs were correctly cloned into pcDNA3.1(+)vector. After a 48-h transfection of HEK293 cells with the recombinant plasmids and G418 selection, the positive cell clones stably expressing the constructs were obtained, which showed expressions of PTHR and DSEL mRNAs detected by RT-PCR. These positive cells showed high levels of PLC and aAMP production in response to PTH stimulation.

CONCLUSIONThe HEK293 cell lines with stable expression of PTH1R or DSEL gene established in this study provide useful cell models for studying the physiological functions of PTH peptides.

Gene Expression ; Genetic Vectors ; HEK293 Cells ; Humans ; Plasmids ; Receptors, Parathyroid Hormone ; genetics ; metabolism ; Sequence Analysis, DNA ; Signal Transduction ; genetics ; Transfection

Objective:To explore the clinical effect of the concept of enhanced recovery after surgery (ERAS) in the perioperative nursing of patients with total hip replacement.Methods:A total of 93 patients with hip osteoarthritis admitted from June 2017 to June 2018 were divided into routine nursing group and ERAS nursing group according to the random number table method. Patients in both groups were treated with unilateral total hip arthroplasty. Routine nursing group was treated with routine perioperative nursing, and ERAS nursing group was treated with ERAS concept. The ERAS nursing team was established, including a head nurse and an orthopedic physician, in addition to the regular nursing staff. The treatment, perioperative pain and nursing satisfaction of the two groups were compared.Results:The first time of getting out of bed activity [(1.95 ± 0.45) days vs. (5.52 ± 2.52) days], the first time of defecation after operation [(1.34 ± 0.56) days vs. (2.15 ± 0.84) days], the time of hospitalization [(5.73 ± 1.65) days vs. (10.04 ± 2.14) days] in ERAS nursing group were shorter than those in routine nursing group, and the cost of hospitalization [(49 450.50 ± 880.52) yuan vs. (53 053.50 ± 780.50) yuan] was lower than that in routine nursing group, the differences were statistically significant ( t=6.639-31.622, P < 0.05). The NRS scores of 24 hours [(3.35 ± 0.85) vs. (4.32 ± 1.05)], 48 hours [(2.65 ± 0.52) vs. (3.48 ± 0.54)] after operation and discharge [(1.74 ± 0.65) vs. (2.36 ± 0.53)] in the ERAS nursing group were lower than those of the routine nursing group, the differences were statistically significant ( t=4.526, 4.856 and 4.456, P < 0.05). The nursing satisfaction rate of ERAS nursing group (95.7%, 45/47) was higher than that of routine nursing group (76.60%, 36/47), the difference was statistically significant ( P<0.05). Conclusion:The idea of ERAS has a good clinical effect in perioperative nursing of patients with total hip replacement.