A method for determination of 6 plant growth regulators by high performance liquid chromatography-tandem mass spectrometry ( HPLC-MS/MS ) coupled with isotope-coded derivatization was developed. d0-10-Methyl-acridone-2-sulfonyl piperazine ( d0-MASPz, light form) and d3-10-methyl-acridone-2-sulfonyl piperazine ( d3-MASPz, heavy form ) were prepared as isotope-coded derivatization reagents for carboxyl compound. The carboxyl plant growth regulator standards and real samples were derivatized by d0-MASPz and d3-MASPz, respectively. The obtained solutions were mixed at a certain ratio, and then injected for HPLC-MS/MS analysis. The light and heavy derivatives were monitored with transitions of [M+H]+m/z 208. 2 and [M+H]+ m/z 211. 2,respectively. With heavy derivative as internal standard for corresponding light derivative, the global isotope internal standard quantification for 6 plant growth regulators was achieved. The results indicated that the proposed isotope-coded derivatization method could provide relative quantitative data with adequate linearity in a 10-fold dynamic range ( R=0 . 9991 ) . The detection and quantitation limits were 0. 19-0. 34 μmol/L and 0. 53-0. 96 μmol/L, respectively. The relative standard deviations were ≤3 . 8%, and the accuracies ranged from 97 . 5% to 103 . 8%.

With comparison of three different methods for the marking of amines compound, an optimal deri vatization method was selected.5-(2-Hydroxyethyl) benzoacridine (HBA) reacts with coupling agent N,N'-carbonyldiimidazole(CDI) to form an activated amide intermediate 2-(benzoacridin) ethyl-imidazole-1-carbox-ylate(BAEIC).BAEIC, which is dual-sensitive probe, reacts preferably with amino compounds at 80 ℃ in the presence of 4-dimethylaminopyridine(DMAP) catalyst in N,N-dimethylformamide(DMF) solvent to give the corresponding sensitively fluorescent derivatives with an excitation maximum at λ_(ex) of 280 nm and an emis sion maximum at λ_(em) of 510 nm.BAEIC-amine derivatives simultaneously exhibited high ionization potential with percent ionization (changing from 5.62% to 58.08% in aqueous acetonitrile and from 2.14% to 56.58% in aqueous methanol.Derivatives were not only sensitive to fluorescence but also to MS ionizable potential.The fluorescence detection limits(5/iV = 3) were 0.12-0.59 μg/L.The online APCI-MS detection limits were 1.9-14 μg/L(S/N=5).

Objective To study the clinical characteristics and to summarize our experience in the diagnosis and treatment of ectopic splenic autotransplantation (ESAP) many years after splenic rupture and splenectomy.Methods The clinical data of 15 patients with ESAP who were treated in the People's Hospital of Zhengzhou from January 1998 to January 2013 were retrospectively analyzed.Results Of the 15 patients,2 patients presented with intestinal obstruction due to the ectopic splenic tissues and required partial resection of small intestine.In 5 patients,the ectopic splenic tissues were in the greater omentum which required resection of part or the whole of the greater omentum.In 4 patients,the ectopic splenic tissues were in the mesentery which required partial or total resection of the tissues.The ectopic tissues were in the pelvis in 3 patients and resection of the left fallopian tube was carried out in 2 patients and resection of the pelvic tissues in 1 patient.In 1 patient the diagnosis was made under laparoscopic biopsy,thus avoiding open surgery.Three patients had multiple implanted sites for these ectopic splenic tissues.Conclusions ESAP is an uncommon disease and it has no clinical characteristics.It is easy to confuse the condition with metastatic deposits.A diagnosis could be made only by combining the history with magnetic resonance imaging,or 99mTc study of red blood cells,or laparoscopic exploration and biopsy.

Objective:To compare the effects of ultrasound-guided dynamic needle tip positioning (DNTP) and long axis in-plane (LAX-IP) techniques for axillary vein puncture and catheterization.Methods:One hundred Society of Anesthesiologists physical statusⅠ-Ⅲ patients of both sexes, aged 18-64 yr, with body mass index of 20-28 kg/m 2, scheduled for elective axillary vein cannulation, were divided into 2 groups ( n=49 each) using the random number table method: DNTP group and LAX-IP group.Axillary vein puncture was performed using DNTP technique and LAX-IP technique under ultrasound guidance in DNTP group and LAX-IP group, respectively.Successful puncture at first attempt, overall successful catheterization, the number of needle tip redirection, and axillary vein puncture time and catheterization time were recorded.The occurrence of complications such as axillary artery puncture, posterior wall penetration of axillary vein, hematoma formation, pneumothorax, and nerve injury was recorded. Results:Compared with group LAX-IP, the success rate of puncture at first attempt was significantly increased, the number of cases required needle redirection was decreased, and the puncture time was shortened ( P<0.05), and no significant change was found in the logarithm of the posterior wall penetration of axillary vein in group DNTP ( P>0.05). No complications such as arterial puncture, hematoma, pneumothorax, or nerve injury occurred in two groups. Conclusions:Compared with LAX-IP technique, ultrasound-guided DNTP technique can dynamically observe the position of the needle tip, the operation is simple and safe, and it is worthy of clinical promotion when used for axillary vein puncture and cannulation.

Objective To evaluate the clinical outcomes and feasibility of multi-modality adjuvant chemotherapy and radiation, which was conducted as postoperative chemotherapy, radiation, and consolidation chemotherapy (CRC) mode for the treatment of advanced endometrial cancer. Methods A retrospective analysis of 124 patients with International Federation of Gynecology and Obstetrics (FIGO) stages Ⅲ and Ⅳ endometrial cancer from Jan. 2004 to Oct. 2012 was conducted in Peking University People′s Hospital and Beijing Obstetrics and Gynecology Hospital. Inclusion criteria were comprehensive staging procedure including hysterectomy, bilateral salpingo-oophorectomy, and (or) selective pelvic aortic lymphadenectomy, and treatment with adjuvant chemotherapy and (or) radiation. The average age of these patients was (55.9 ± 8.4) years old (range from 23 to 79 years old). According to different postoperative adjuvant treatment modes, the patients were divided into CRC group, chemotherapy-radiotherapy (CR) group and single chemotherapy (C) group. The survival and side effects of the three groups were compared. Results (1) One hundred and twenty-four patients with advanced stage endometrial cancer were identified and received postoperative adjuvant therapies.Sixty-one (49.2％, 61/124) cases of them received postoperative CRC fashion, 19 (15.3％, 19/124) received postoperative CR and 44 (35.5％, 44/124) cases received C. The age, stage, grade and type of surgery of the three groups were not significantly different (all P>0.05);while, the pathology, chemotherapy cycles and chemotherapy regimens differed significantly (all P<0.05). (2) The progression-free survivals (PFS) of the patients with CRC, CR, and C group were (121±7), (68± 15), and (100±11) months, respectively. The 3-year PFS rates were 87.9％, 43.7％, and 61.4％, respectively. The 5-year PFS rates were 82.2％, 36.4％, and 61.4％, respectively. The above indicators were significantly higher in the CRC group than in the CR group (all P<0.01), and there was no difference between the CRC group and the C group (P=0.037). The overall survival (OS) of patients with CRC, CR, and C group were (128 ± 6), (80 ± 12), and (99 ± 10) months, respectively. The 3-year OS rates were 87.8％, 72.4％, and 67.1％, the 5-year OS rate were 84.2％, 54.3％, and 64.1％, respectively. The above indicators were significantly higher in the CRC group than those in the CR group and C group (all P<0.01). (3) There was no difference in the frequency of adverse effects either chemotherapy, such as severe bone suppression or radiotherapy;hepatotoxicity,blood transfusion, dose modifications;or cycle delays between the CRC, CR and C group (all P>0.05). (4) In the univariate analysis shown that, stage, the fashion of postoperative adjuvant therapy and type of surgery were risk factors for tumor progression in patients with advanced endometrial cancer (P<0.05). After adjusted for FIGO stage and type of surgery, the tumor progression hazard ratio (HR) was 3.931 (95％CI:1.734-8.914, P=0.001) for the CR group and 2.188 (95％CI:1.010-4.741, P=0.047) for the C group, compared to the CRC group. Conclusion Sequential CRC delivered in a"sandwich"fashion for the treatment in advanced endometrial cancer could significantly improve the 3-year and 5-year OS rates and have a similar adverse effect profile compared with other sequencing modalities.

The Guarding field theory is a unique theoretical system of traditional Chinese medicine for the treatment of surgical diseases such as scabies, sores and gangrene. The hooping in place is the best performance of Guarding field theory. This paperreviews the research progress of Guarding field theory from the aspects of theoretical research, basic research, clinical research in recent years, which shows the connotation of this theory has been expanded. and the generalized concept of Guarding field theory refers to human's healthy Qi, and the field theory has been widely used in treating diseases likemalignant tumor, postoperative anorectal infection, refractory wound to guide clinical treatment.

c-Myc protein encoded by c-Myc (cellular-myelocytomatosis viral oncogene) gene regulates the related gene expression through the Wnt/β-catenin signaling pathway, and has received extensive attention in recent years. The purpose of this study was to express Helicoverpa armigera c-Myc gene (Ha-c-Myc) by using prokaryotic expression system, prepare the polyclonal antibody, examine the spatio-temporal expression profile of Ha-c-Myc, and investigate the possible function of Ha-c-Myc in regulating H. armigera sterol carrier protein-2 (SCP-2) gene expression. The Ha-c-Myc gene was amplified by PCR and cloned into a prokaryotic expression plasmid pET-32a(+). The recombinant plasmid pET-32a-Ha-c-Myc was transformed into Escherichia coli BL21. IPTG was used to induce the expression of the recombinant protein. Protein was purified by Ni²⁺-NTA column and used to immunize New Zealand rabbits for preparing the polyclonal antibody. The Ha-c-Myc expression levels in different developmental stages (egg, larva, prepupa, pupa, and adult) of H. armigera and different tissues (midgut, fat body, head, and epidermis) of the prepupa were determined by real-time quantitative reverse transcription PCR (qRT-PCR). Ha-c-Myc siRNA was synthesized and transfected into H. armigera Ha cells. The relative mRNA levels of Ha-c-Myc and HaSCP-2 in Ha cells were detected by qRT-PCR. Results showed that the pET-32a-Ha-c-Myc recombinant plasmid was constructed. The soluble Ha-c-Myc protein of about 65 kDa was expressed in E. coli. The polyclonal antibody was prepared. Western blotting analysis suggested that the antibody had high specificity. Enzyme linked immunosorbent assay (ELISA) showed that the titer of the antibody was high. Ha-c-Myc gene expressed at all developmental stages, with high levels in the early and late instars of larva, and the prepupal stage. Tissue expression profiles revealed that Ha-c-Myc expressed in various tissues of prepupa, with high expression level in the midgut, but low levels in the epidermis and fat body. RNAi results showed that the knockdown of Ha-c-Myc expression significantly affected transcription of HaSCP-2, leading to a 50% reduction in HaSCP-2 mRNA expression level. In conclusion, the Ha-c-Myc was expressed through a prokaryotic expression system, and the polyclonal anti-Ha-c-Myc antibody was obtained. Ha-c-Myc may promote the expression of HaSCP-2 and play an important role in the lipid metabolism of H. armigera. These results may facilitate further study on the potential role and function mechanism of Ha-c-Myc in H. armigera and provide experimental data for exploring new targets of green pesticides.
Animals ; Rabbits ; Escherichia coli/metabolism* ; Enzyme-Linked Immunosorbent Assay ; Moths/genetics* ; Blotting, Western ; Larva/genetics* ; Isoantibodies/metabolism* ; Antibody Specificity