Objective: To screen and identify oxaliplatin-resistance-associated proteins in CRC (colorectal cancer) cell lines using proteomics technologies in order to find new biomarkers for individual therapy of CRC. Methods: Oxaliplatin-resistant human CRC cell line HT-29/L-OHP (oxaliplatin) was established. The total proteins in HT-29 and HT-29/L-OHP cells were extracted. The differentially expressed proteins between HT-29 and HT-29/L-OHP cells were screened and identified using 2-DE (two-dimensional gel electrophoresis) and MALDI-TOF-MS (matrix assisted laser desorption-ionization time-of-flight tandem mass spectrometry). Some proteins obtained were validated by Western blotting. Results: The 2-DE maps of total proteins in HT-29 and HT-29/L-OHP cells were established. Of the 38 protein spots identified as differentially expressed proteins (over two-fold, P < 0.05) between HT-29 and HT-29/L-OHP cells, 37 protein spots were positively identified by MALDI-TOF-MS (17 proteins were up-regulated and 20 proteins were down-regulated as compared with the parental HT-29 cells). The result of Western blotting showed that the PCBP1 [poly (C)-binding protein-1] and TUBB2A (tubulin beta 2A ) proteins were up-regulated while ANXA3 (annexin A3) and STIP1 (stress-induced-phosphoprotein 1) proteins were down-regulated in HT-29/L-OHP cells. The result of Western blotting was consistent with that of proteomics. Conclusion: There were 37 oxaliplatin-resistance-associated proteins in CRC identified in this study which may provide useful evidence in further research on mechanism of oxaliplatin-resistance in CRC. Copyright © 2013 by TUMOR.

Objective To silence Bax gene in H9c2 cells by ultrasound-targeted microbubble destruction(UTMD) combined with polyethylenimine.Methods The shRNA expression plasmid targeting Bax gene was co-treated with microbubbles and polyethylenimine.Then the mixture added to H9c2 cells were irradiated by ultrasound.Grouping:① plasmid group;② Lipofectamine2000 + plasmid group; ③ UTMD + plasmid group;④ PEI + UTMD + plasmid group;⑤ PEI + SonoVue + plasmid group (without ultrasound irradiation).The transfection efficiency was assessed by fluorescence microscope and FCM.The expression of Bax mRNA and protein were detected by RT-PCR and Western Blot.Results Bax shRNA wasn't destroyed by ultrasound exposure.The transfection efficiency of PEI + UTMD + plasmid group was higher than any other groups obviously (P ＜0.05).The result of RT-PCR and Western Blot showed inhibition efficiency of Bax mRNA and protein expression in PEI + UTMD + plasmid group were (69.41 ± 4.91) ％ and (64.09 ± 2.38)％ separately,which were higher than any other groups significantly (P ＜0.05).Conclusions UTMD combined with polyethylenimine can inhibit the expression of Bax gene in H9c2 cells effectively.

Objective To investigate the significance and effect of ultrasonic diagnosis on the autologous bone marrow mesenchymal stem cells (MSC) in angiogenesis. Methods Twenty-four New Zealand rabbits were divided into experiment group (12) and the control group (12). Then rabbit bone marrow MSCs from experiment group were isolated, caltured and marked with Brdu. After ischemic hind limb animal model on all rabbits was set up, autologous bone marrow MSCs were directly injected into the ischemic hind limb muscles in experiment group while same volume normal saline was used in the control group. Two weeks after the implantation of autologous bone marrow MSCs, 2D and color Doppler flow imaging (CDFI) detection were used in rabbit femoral artery of the two groups to observe the inner diameter of the blood vessel, the peak velocity and the acceleration time. The disposition of transplaned cells and the state of angiogenesis in ischemic muscles were assessed using immunofluorescence staining. Results The results of 2D and Doppler ultrasound detection showed the inner diameter of the blood vessel and the peak velocity of the blood current in experiment group obviously higher than that of the control group , and the acceleration time was obviously smaller than that of the control group P<0.01. The immunofluorescence staining showed there were transplanted cells existed in transplanted portion and state of angiogenesis was supurior obviously than that of the control. Conclusions Bone marrow MSCs had the effect to promote angiogenesis. Implantation of autologous bone marrow MSCs was a simple and efficient therapeutic method for the ischemia hind limb. Using high-frequency ultrasound to detect femoral artery may provide a practical and useful method to evaluate the effect on implanted autologous bone marrow mesenchymal stem cells.

BACKGROUNDCerebral amyloid angiopathy (CAA) is one of the main causes of spontaneous intracranial hemorrhage (ICH). No established link is available between pathological scores of CAA and its outcome. This study aimed to identify the correlations between pathological severity and poor postoperative outcome in the Chinese population.

METHODSBetween May 2006 and April 2011, 367 consecutive patients who underwent surgery for CAA-related ICH in 71 hospitals throughout the mainland of China were enrolled in this study. Twelve months after surgery, we evaluated these patients' outcomes according to the modified Rankin Scale (mRS) and statistically correlated risk factors (demographics, medical history, pathological results, and surgical details) that are associated with a favorable (mRS < 3) and poor (mRS ≥ 3) outcome groups.

RESULTSRisk factors for poor postoperative outcome in 367 patients with CAA-related ICH included advanced age (OR 1.034, 95%CI 1.001 - 1.067, P = 0.042), CAA pathology severity (OR 2.074, 95%CI 7.140 - 16.25, P < 0.001), lobar hematoma (OR 0.225, 95%CI 0.104 - 0.486, P < 0.001), presence of intraventricular hemorrhage (OR 0.478, 95%CI 0.229 - 1.001, P = 0.050), and/or subarachnoid hemorrhage (OR 2.629, 95%CI, 1.051 - 6.577, P = 0.039).

CONCLUSIONSPoor postoperative outcome of patients with CAA-related ICH was more related to the severe pathological manifestation instead of other factors. Prior ischemia may present an early stage of CAA.

Aged ; Cerebral Amyloid Angiopathy ; pathology ; physiopathology ; China ; Female ; Humans ; Intracranial Hemorrhages ; pathology ; physiopathology ; Male ; Middle Aged ; Risk Factors

OBJECTIVETo analyze the clinical features of adrenal metastasis.

METHODSFrom January 1993 to December 2004, 103 cases of adrenal metastasis were reviewed.

RESULTSLung and hepatocellular carcinoma were the most common primary tumor of adrenal metastatic tumor, which about 36.9% (38/103) and 42.7% (44/103) of all cases, followed by renal carcinoma 6.8% (7/103), colorectal carcinoma 4.9% (5/103), stomach carcinoma 3.9% (4/103), breast cancer 1.9% (2/103), unknown primary tumor 2.9% (3/103). Most of these were low differentiation. The mean diameter of adrenal metastasis was 3.9 cm. The mean interval from detection of primary tumor to adrenal metastasis was 9.5 months. And 79.6% (82/103) were detected as a part of multiorgan metastasis. Only 5 cases (4.9%) were presented with pain in the back. There was little characterization of ultrasonography, CT and MRI, color-Doppler and selective arterial imaging showed little blood supply. All of patients were treated with synthetic methods, 16 cases (15.5%) who had undergone adrenalectomy for metastasis disease had a improved survival compared with those non-adrenalectomy.

CONCLUSIONSThere is no particular presentation of clinic and imaging, diagnosis depending on history, follow-up and the pathological presentation of primary tumor. There are no standard treatment guidelines for this group of patients. When the primary tumor could be resected or be well controlled, and there is no other evidence of metastasis, adrenalectomy is recommended. Transarterial chemoembolization (TACE) could not actually be performed.

Adrenal Gland Neoplasms ; diagnosis ; secondary ; therapy ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; pathology ; Combined Modality Therapy ; Female ; Humans ; Liver Neoplasms ; pathology ; Lung Neoplasms ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Survival Analysis ; Treatment Outcome

OBJECTIVETo investigate the clinical manifestations, EB virus serology and treatment outcome of nasopharyngeal adenocarcinoma (NPAC).

METHODSClinical records of NPAC patients between 1964 and 2000 in Cancer Center of Sun Yat-sen University were retrospectively reviewed.

RESULTSAmong 48 patients with NPAC, 45.2% (7 cases of N1, 8 cases of N2 and 4 cases of N3) of them presented with cervical metastasis. Pathologically, common type and salivary gland type of NPAC accounted for 58.3% (28 cases) and 41.7% (20 cases) respectively. The positive rate of the EB virus antibody VCA-IgA was 56.7% in the whole group and only 23.7% in the salivary gland type of NPAC. The overall local control rate and the 5-year disease free survival rate by Kaplan-Meier method were 87.0% (40/46) and 65.2% respectively. Baseline data analysis showed that age, gender, N stage and M stage were not the significant factors, never the less the T stage was not balanced between the two groups (surgery plus radiotherapy vs radiotherapy alone, chi2 = 4.801, P = 0.045). The patients treated by surgery plus radiotherapy had significantly higher 5-year disease free survival rate than by radiotherapy alone (88.9% vs 74.7%, Log Rank test: chi2 = 4.272, P = 0.039). Cox's multivariate analysis showed treatment modality and N stage were the significant factors influencing survival (RR were 15.276 and 6.529, P < 0.05).

CONCLUSIONSNPAC is a distinct entity in all types of nasopharyngeal carcinoma. EB virus serology has limited value in its diagnosis. Surgery plus radiotherapy could be another choice of treatment for early lesions of NPAC.

Adenocarcinoma ; diagnosis ; pathology ; virology ; Adolescent ; Adult ; Aged ; Antibodies, Viral ; analysis ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnosis ; pathology ; virology ; Prognosis ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the incidence of lymph node metastasis (LNM) in the right para-tracheal triangle (RPT) of esophageal carcinoma patients and the technique of dissection.

METHODSOn the top of double mediastinal and abdominal lymphadenectomy, 333 esophageal carcinoma patients received RPT lymphadenectomy through the right pleural apical approach from 1990 to 2001.

RESULTSIn these 333 patients, the lymph node metastasis (LNM) rate in the RPT was 36.40%. A total of 457 nodes among 2 159 nodes removed gave a metastasis degree of 24.96%. The LNM rates in RPT for cervical, upper third, middle third, and lower third segments of esophagus were 66.67%, 45.45%, 34.19% and 15.79% (P < 0.05), while their respective metastasis degrees were 44.44%, 27.04%, 24.32% and 18.92% (P > 0.05). The frequency of positive nodes in the RPT for PTI, PT1, PT2, PT3 and PT4 was 0, 17.24%, 28.7%, 45.16% and 53.57%, while those of metastasis degree were 0, 8.77%, 17.62%, 33% and 41.17% (P < 0.01). The frequency of LNM in the RPT in papillary, erosive, patch-like and covert type of early tumor was 40%, 3.85%, 0 and 0 (P < 0.05), while those of the metastasis degree were 29.41%, 1.82%, 0 and 0 (P < 0.01). Higher rate of LNM in progressive stenotic esophageal carcinoma was observed compared with those of the other gross types (56.52%, P < 0.05), so was the degree (P < 0.01). The frequency of LNM in the RPT for mono-focal and multi-focal tumor was 34.98% and 70% without significant difference (P > 0.05), while the degree was 24.29% and 53.33% (P < 0.05). Postoperative complications were: leak (0.6%), and recurrent laryngeal nerve injury (1.2%). No injury of vein or infra-clavicular artery, tracheal damage or mortality occurred.

CONCLUSION1. The lymph node metastasis from esophageal carcinoma has a tendency of wide spread and right para-tracheal triangle is an important region to be doomed. 2. With location, depth of tumor invasion and differentiation of tumor as major factors affecting LNM of esophageal carcinoma, dissection of this region should be paid more emphasis. 3. In early lesions, higher frequency of LNM in the RPT is found in papillary and erosive lesions than in the other macroscopic types. 4. Exposing the RPT, lymph node by dissection through a right pleural apical approach is very important and significant.

Adult ; Aged ; Cardia ; Esophageal Neoplasms ; pathology ; surgery ; Esophagectomy ; methods ; Esophagus ; pathology ; Female ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Mediastinum ; Middle Aged ; Neck ; Neoplasm Invasiveness

OBJECTIVETo assess the binding activity of polypeptide containing human Na+, K+-ATPase alpha1 subunit M1-M2 extracellular segment (HES1 derivative).

METHODSHES1 derivative was synthesized by Fmoc method and purified by high-performance liquid chromatography-mass spectrometry, and its binding activity was identified by radioligand binding assay.

RESULTS3H-ouabain and synthetic HES1 derivative showed some binding activity with the equilibrium dissociation constant (KD) of 24.58 nmol/L, with the the receptor density of 492.43 fmol x mg(-1) pro. and IC50 of 3.078 x 10(-7) mol/L.

CONCLUSIONHES1 derivative can bind to ouabain and has the potential of becoming an effective therapeutic agent.

Binding Sites ; drug effects ; Extracellular Space ; metabolism ; Humans ; Ouabain ; chemistry ; pharmacology ; Peptides ; chemistry ; Protein Binding ; Sodium-Potassium-Exchanging ATPase ; chemistry ; genetics ; metabolism

OBJECTIVETo study the impact of an agonist anti-CD(40) monoclonal antibody 5C11 on the induction and biological characteristics of leukemic dendritic cells.

METHODSCombinations of 5C11 and different cytokines were used to induce differentiation of leukemic blasts into dendritic cells. Morphology was observed by light microscopy. Surface antigens of the induced cells were analyzed by fluorescence-activated cell sorting (FACS), the yields of dendritic cell by cell counting, the levels of IL-6 and IL-12 by ELISA, T cell proliferating activity by allo-mixed lymphocyte reaction (MLR) in vitro. Allogeneic T cells were stimulated with leukemic dendritic cells and T-cell cytotoxicity was measured by MTT assay.

RESULTSWhen cultured with combinations of 5C11 and different cytokines, the leukemic cells isolated from the patients could differentiate into dendritic cells. The morphology showed typical features of dendritic cells, which expressed high levels of CD(40), CD(80) and CD(86). In comparison with the original leukemia cells, the leukemic dendritic cells secreted less IL-6 but more IL-12 (P < 0.05). The leukemic dendritic cells were potent to stimulate the proliferation of allogeneic T cells, and the latter was able to lyse the original leukemia cells.

CONCLUSIONLeukemic blasts could be induced to differentiate into functional dendritic cells. It may be of great value in the adoptive immunologic therapy of leukemia.

Antibodies, Monoclonal ; immunology ; CD40 Antigens ; physiology ; Cell Differentiation ; Dendritic Cells ; immunology ; Humans ; Immunophenotyping ; Immunotherapy ; Interleukin-12 ; biosynthesis ; Interleukin-6 ; biosynthesis ; Leukemia ; immunology ; pathology ; therapy

OBJECTIVETo discuss the impacts of moxibustion for regulating spleen and stomach function on the survival quality of the patients of end stage renal disease (ESRD) with maintenance hemodialysis (MHD).

METHODSOne hundred and nine cases of uremia with MHD from 3 hemodialysis centers were randomized into an observation group (58 cases) and a control group (51 cases). The regular hemodialysis and conventional medication were used in the two groups. In the observation group, on the basis of the common treatment, moxibustion was applied to Zusanli (ST 36) and Sanyinjiao (SP 6), 2-3 times a day, the treatment of 4 weeks made one session. Totally, 3 sessions were required and the follow-up lasted for 3 months. KDQOL-SF (kidney disease quality of life short form,KDQOL-SFTM 1. 3) was adopted for the questionnaire investigation on survival quality before treatment, after treatment and at the end of follow-up separately in the two groups.

RESULTSAfter treatment, the survival quality scores in terms of physical functioning (83.62+/-13.27 vs 79.32+/- 22. 17), general health (58. 88+/- 20.24 vs 48.82+/-20.89) and vitality (77.07+/-15.56 vs 70. 59+/-22.61) in the observation group were higher than those in the control group (all P<0. 05). In comparison before and after treatment in the same group, the survival quality scores in terms of physical functioning, general health, vitality and symptoms/problems were all improved in the observation group (all P<0. 05). At the end of follow-up, the survival quality scores in terms of physical functioning, general health, mental health, social functioning, vitality, effects of kidney disease and cognitive function were higher in the observation group as compared with those in the control group (all P<0. 05). In comparison of the results at the end of follow-up with those before treatment, the survival quality scores in terms of vitality, symptoms/problems and cognitive function in the observation group were improved (all P< 0. 05). The differences were not significant in all of the 19 fields of survival quality evaluation before and after treatment, and after follow-up in the control group (all P>0. 05).

CONCLUSIONMoxibustion for regulating spleen and stomach function improves the survival quality of the patients with hemodialysis in terms of physical functioning, general health and vitality, which benefits the psychological condition of the patients, resulting in the improvements of the survival quality in the fields of mental health, social functioning, effects of kidney disease and cognitive function.

Adult ; Aged ; Female ; Humans ; Kidney Failure, Chronic ; physiopathology ; therapy ; Male ; Middle Aged ; Moxibustion ; Quality of Life ; Renal Dialysis ; Spleen ; physiopathology ; Stomach ; physiopathology