To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.
Amides ; chemistry ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Rhizome ; chemistry

Objective To study the sources and the relationship between the management and the outcome of hemorrhage after cephalic pancreatoduodenectomy.Methods The clinical data of 370 patients who underwent pancreatic resection at the Lihuili Hospital and the Second Affiliated Hospital of Zhejiang University were retrospectively analyzed.Results Postoperative bleeding occurred in 35 patients with 11 deaths.Among those intraabominal bleeding occurred in 14 cases and gastrointestinal hemorrhage occurred in 22,with one case suffering from both.Bleediug developing within 72 hours after operation in 12 cases (early-stage group),which was caused by improper intraoperative homeostasis.In other 23 cases,bleeding 72 hours after operation(later stage group)was caused by the erosion following pancreatic and/or bile leakage.Relaparotomy was performed in 13 cases and endoscopic homeostasis was performed in 3. Relaparotomy or endoscopic homeostasis was superior to that of conservative therapy in the early-stage group (P0.05).Pancreatic or bile leakage was identified as the significant risk factors for the postoperative bleeding.Conclusions In order to prevent the postoperative hemorrhage and to reduce the mortality of pancreatic resection,skillful techniques,expeditious homeostasis,proper management of stump pancreas and the prevention of pancreatic and bile leakage are essential.

Animals ; Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus ; genetics ; Rats

OBJECTIVETo investigate the structure and function of the N-terminal region (NTR) of death receptor 5 (DR5).

METHODSA series of deletions of the DR5 extracellular domain (DR5-ECD) proteins were expressed in E.coli. and purified by affinity chromatography. The binding ability of these deletant proteins to AD5-10, a mouse anti-human DR5 monoclonal antibody, was evaluated by immunoblotting and ELISA.

RESULTSRecombinant DR5-ECD proteins containing the NTR were recognized and bound by AD5-10, while the other deletant proteins without the NTR failed to interact with AD5-10.

CONCLUSIONThere is an AD5-10 targeting site in the NTR of DR5, which may play a role in developing novel immunotherapies for cancers.

Animals ; Antibodies, Monoclonal ; chemistry ; Binding Sites ; Gene Deletion ; Genetic Engineering ; Genetic Vectors ; Humans ; Mice ; Protein Binding ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; chemistry ; genetics ; metabolism

Objective To investigate the effect of 131I on apoptosis of thyrocytes in patients with Graves disease. Methods Forty-seven patients with Graves disease were divided into two groups, two week group (G2w) and four week group (G4w). All patients underwent thyoid needle biopsy before 131I treatment and the repeated biopsy at two weeks (G2w) or four weeks (G4w) after 131I treatment. The positive units of pro-apoptotic proteins (Fas, FasL) and anti-apoptotic protein (Bcl-2) were studied with immunohistochemistry staining. The differences of the two groups were compared with t-test. Liner correlation analysis was applied to study the correlation between 131I dose and apoptosis-related proteins and that between serum sTSH after 131I treatment and apoptosis-related proteins. Results Fas, FasL and Bcl-2 expression (positive units) were significantly increased in both groups after 131I treatment, G2w :22.84 ± 9.31 vs 16.20 ± 6.75,21.13±6.29vs 14.56±4.06, 21.69±7.83 vs 15.22 ±5.94, t= -3.08, -3.73, -4.05 (allP<0.05); G4w:21.69 ±4.52 vs 15.83 ±5.03, 19. 11 ±3.75 vs 14.02 ±4.98, 19.06 ±3.44 vs 16.63 ±4. 73, t = - 5.26, - 5.00, - 2.41 (all P<0.05). However, no statistical differences were found between G2w and G4w (t = 0. 53, 0. 82, 1.46, all P > 0.05). Significant correlation was found between 131I 0. 727, rFasL = 0. 763 (both P<0.05)), but not between the dose and Bcl-2, rBcl-2 = - 0. 094, 0. 102(both P > 0.05). There were significant correlation between serum sTSH three months after 131I treatment and apoptosis-related proteins, rFas = 0.433, rFasL = 0. 601, rBcln2 = - 0. 397, (all P<0. 05). Conclusions 131I can induce thyrocytes to express the pro-apoptotic proteins in patients with Graves disease.

OBJECTIVETo study the effect of Wenhua Juanbi Recipe (WJR) on proliferation and apoptosis of synoviocytes in rats with collagen-inducing arthritis (CIA).

METHODSA CIA model was induced by intradermal injection of bovine collagen type II emulsion at the base of rat tails. Thirty modeled healthy Wistar rats were randomly assigned to one of three groups (10 per group): the model group, the methotrexate (MTX)-treated group (0.78 mg/kg) and the WJR-treated group (22.9 g/kg). A group of 10 healthy rats was used as normal control. Treatments or normal saline for the control group were administered by oral gavage once daily. Rats were sacrificed after 30-day treatment and subjected to the following examinations: arthritis index (AI) was estimated, inflammatory cell infiltration and proliferation in synovial membrane were evaluated by microscopy, the synoviocyte apoptosis was determined by TUNEL assay, and the cell apoptosis index was calculated.

RESULTSAI was lowered significantly in the WJR group compared to the model group (P<0.01). The pathological findings observed in the model group were reversed in the WJR group, including increase in inflammatory cell infiltration and synoviocyte proliferation in synovial membrane and reduction in cell apoptosis index (all P<0.01).

CONCLUSIONSSynoviocyte proliferation and apoptosis reduction were present in CIA rats. WJR was effective in treating the rat model of CIA. The therapeutic effect might be exerted through inducing apoptosis and suppressing proliferation of synoviocytes.

Animals ; Apoptosis ; drug effects ; Arthritis, Experimental ; pathology ; Cattle ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Wistar ; Synovial Membrane ; drug effects ; pathology

OBJECTIVETo investigate the changes of encephalic contents of somatostatin (SS) and arginine vasopressin (AVP) in rats with vascular dementia.

METHODSTwenty-four male Sprague-Dawley rats were randomly divided into three groups: vascular dementia group (VDMG), sham operation group (SOG) and control group (CG). The vascular dementia model was established by permanent bilateral vertebral artery occlusion through electric coagulation and shut-off the bilateral carotid arteries. The remember behavior of animals was tested and the contents of SS and AVP in various encephalic region (frontal cortex, temporal lobe, hippocampus, cerebral ganglion and corpora striatum) were determined with radioimmunoassay.

RESULTDuring the 15-day-long remembering test, the frequency of making mistakes in the VDMG was higher remarkably than that in SOG and CG (P<0.05); and the relative contents of SS were decreased in frontal area cortex, temporal lobe, hippocampus, cerebral ganglion and corpora striatum (P<0.01), while decrease of AVP contents was only detected in temporal lobe and corpora striatum (P<0.05).

CONCLUSIONThe disturbance of learning and memory function might be associated with SS and AVP after multiple cerebral infarction in the animals.

Animals ; Arginine Vasopressin ; metabolism ; Brain ; metabolism ; Dementia, Vascular ; metabolism ; physiopathology ; Male ; Memory ; physiology ; Neuropeptides ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Somatostatin ; metabolism

OBJECTIVETo detect the expression profile of NK ligands in acute leukemia cell lines and investigate the differential expression pattern between acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML).

METHODSUsing quantitative real-time PCR, 23 NK ligands (MICA, MICB, ULBP-1, ULBP-2, ULBP-3, ULBP-4, HLA-E, HLA-G, CD48, NBTA, HLA-F, LLT-1, PVR, Nectin2, CD72, CD80, ICAM-1, LFA-3, CRACC, Fas, DR4, DR5, TNFR1) were detected in 6 acute leukemia cell lines, including 3 ALL cell lines (CEM, Jurkat T, Reh) and 3 AML cell lines (HL-60, KG-1a, NB4), respectively. Independent-samples t test analysis was performed to determine statistical significance.

RESULTSUsing β-actin as reference gene, the relative expression results showed that the expression of 4 NK ligands between ALL and AML is significantly different. Specifically, the level of ULBP-2 is higher in ALL (CEM: 1, Jurkat T: 0.617, Reh: 0.246) than that in AML (HL-60: 0.000, KG-1a: 0.003, NB4: 0.000)(P = 0.047). However, the expressions of CD48, PVR(PVR-1, PVR-2) and DR4 is higher in AML (HL-60: 13.987, 4.403, 10.334, 8.711; KG-1a: 5.387, 2.900, 7.315, 4.512; NB4: 7.763, 3.248, 7.049, 6.127) than that in ALL (CEM: 1, 1, 1, 1; Jurkat T: 2.035, 1.553, 3.888, 0.449; Reh: 1.559, 0.000, 0.000, 1.304) (P = 0.044, 0.014, 0.014, 0.011). And there're no significant differences between the rest 19 NK ligands.

CONCLUSIONSULBP-2, CD48, PVR and DR4 might play an important role in the distinct mechanisms in leukemogenesis between ALL and AML and could be potential targets for diagnosis and treatment.

Acute Disease ; Antigens, CD ; genetics ; metabolism ; CD48 Antigen ; Cell Line, Tumor ; GPI-Linked Proteins ; genetics ; metabolism ; HL-60 Cells ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Leukemia ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Ligands ; Membrane Proteins ; genetics ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; genetics ; metabolism ; Receptors, Virus ; genetics ; metabolism

OBJECTIVETo explore the optimal methods for the reconstruction and preservation of the glans after partial penis resection in the treatment of early-stage penile cancer.

METHODSBetween January 2012 and June 2015, we treated 6 cases of early- stage penile cancer by partial penis resection, inner thigh skin graft, and glans reconstruction and followed them up for 0.5-3 years.

RESULTSThe length of the penis before and after operation was ([6.5 ± 1.2] vs [4.5 ± 1.8] cm) in the flaccid state and ([12.8 ± 2.3] vs [9.1 ± 2.1] cm) in the erectile state. The sense of the reconstructed glans was completely recovered at 3 months after surgery. The glans skin was pale red and soft, nearly normal at 12 months, with no obvious graft contracture or scar formation. All the patients achieved normal erection and their partners were satisfied with their intercourse. No recurrence or metastasis was observed.

CONCLUSIONThe strategy of partial penis resection, inner thigh skin graft and glans reconstruction, simple, effective, and with few complications, is one of the best treatments of early-stage penile cancer, which not only ensures radical removal of the tumor but also maximally reserves the function of the organ.

Humans ; Male ; Penile Neoplasms ; surgery ; Penis ; surgery ; Reconstructive Surgical Procedures ; Skin Transplantation ; Thigh

OBJECTIVETo trace magnetically labeled MSCs transplanted into the rat livers by magnetic resonance imaging (MRI).

METHODSFeridex and DAPI labeled rat mesenchymal stem cells (MSCs) were injected via portal veins into carbon tetrachloride treated rats. MRI was performed with a clinical 1.5 T MRI machine immediately before the MSCs injection and at h 1, d 3, d 7, and d 14 after the injection, and then the signal-to-noise ratio (SNR) was measured. MRI findings were compared with the liver histopathologies after the slides were stained with fluorescence dye and Prussian blue.

RESULTSThe SNR for liver was 1.10+/-0.26 at hour 1, 8.18+/-1.55 at day 3, 11.08+/-1.30 at day 7, and 14.15+/-1.02 at day 14 respectively. Within 7 days after the MSCs transplantation, the SNRs of the livers were significantly lower than those before the transplantation (P less than 0.05). Histologically, the blue fluorescent particles under the fluorescence microscopy matched in distribution with the iron particles on the Prussian blue stained slides.

CONCLUSIONThe magnetically labeled MSCs transplanted into livers give rise to an obvious signal decrease, and can be tracked with a 1.5 T clinical MRI machine for up to 7 days after MSCs transplantation.

Animals ; Image Enhancement ; methods ; Liver ; pathology ; Magnetic Resonance Imaging ; Male ; Mesenchymal Stem Cell Transplantation ; Radioactive Tracers ; Rats ; Rats, Wistar