Objective The aim of our study was to develop a robust LC-MS/MS method for determination of MN and NMN in blood plasma.Methods A liquid chromatography -tandem mass spectrometric ( LC-MS/MS) method was used, with signal linearity, lower limits of quantitation, precision and accuracy being evaluated.The study recruited 126 healthy volunteers, and MN and NMN in blood plasma were determined.At the same time samples from 21 patients ( 17 pheochromocytoma, 4 ectopic pheochromocytoma) , a hypertension group of 108 persons, and a control group of 84 persons were analyzed. A paired T test was used to compare the MN and NMN levels between the different groups.Results The performance characteristics for the method in terms of linearity, lower limits of quantitation, precision and accuracy were verified.Significant differences were found between the concentration levels of MN and NMN in the diseased and healthy groups.Conclusion A robust and reliable LC-MS/MS method for the determination of MN and NMN in blood plasma has been developed and was shown to be suitable for clinical application.

Objective We are going to establish a robust liquid chromatography-tandem mass spectrometric(LC-MS/MS) method for plasma aldosterone assay.Methods 324 healthy individuals were enrolled in Zhongshan Hospital from February to April in 2016 for reference interval survey.The signallinearity,lower limits of quantitation,precision and accuracy of LC-MS/MS have been evaluated.Results from LC-MS/MS and RIA methods were compared.Software SPSS17.0 software was used for statistical analysis.Results The performance characteristics for the method in terms of linearity,lowerlimits of quantitation,precision and accuracy were verified.Linear range of ALD were between 25-2000 pg/ml;the LC-MS/MS assay had a limit of quantitation of 20 pg/ml for ALD;the intra-and inter-assay CV of ALD were ＜10％ and ＜6％,respectively;the recovery of ALD from serum samples ranged between 97.3 and 105.8％ The reference value of ALD in health people ranged between 21-211.6 pg/ml The regression equation by LC-MS/MS (X) and RIA (Y) was:Y =0.271X + 138.900(r=0.43;n=322).Conclusion LC-MS/MS method is robust and reliable for the analysis of aldosterone in plasma and suitable for clinical application.

OBJECTIVETo investigate the feasibility and short-term efficacy of laparoscopic resection of primary localized gastric gastrointestinal stromal tumors (GIST) by comparing with open surgery.

METHODSClinicopathological data of 167 gastric GIST patients undergoing operation in Zhongshan Hospital from June 2008 to December 2013 were retrospectively analyzed, among whom 55 received laparoscopic surgery and 112 underwent open surgery for primary local gastric GIST. Efficacy of different size and different location of GIST was compared between laparoscopic and open groups.

RESULTSThere was no conversion to open surgery in laparoscopy group. Compared with open surgery, laparoscopic resection for gastric GIST smaller than 5 cm or located at anterior wall, greater curvature, lesser curvature, was associated with similar operation time(P>0.05), but less blood loss, shorter post-hospital stay or flatus time(all P<0.05). The operative outcomes were similar between laparoscopic and open resection for gastric GIST bigger than or equal to 5 cm or located at posterior wall(all P>0.05), except the longer operation time in laparoscopy group(P<0.05). The incidence of postoperative complication did not differ between two groups. Laparoscopic group had 2 patients with gastroparesis and open group had 2 gastroparesis, 2 pulmonary infection, and 1 poor wound healing(all P>0.05), which all recovered after conservative treatment. During 7 to 84 months(median 35) of follow-up, no recurrence or hepatic metastasis was found in laparoscopy group, and 3 hepatic metastases in open group. There was no significant difference of recurrence-free survival between two groups(P>0.05).

CONCLUSIONLaparoscopic resection for gastric GIST is safe and effective in selected patients, especially for those with tumors smaller than 5 cm, or located at anterior wall, greater curvature, lesser curvature, whose short-term outcomes are better than open resection.

OBJECTIVETo explore the clinicopathological characteristics, efficacy, and prognostic factors for patients with duodenum gastrointestinal stromal tumor(GIST).

METHODSClinicopathological and follow-up data of 80 patients with duodenum GIST in the Zhongshan Hospital from January 2000 to December 2013 were analyzed retrospectively.

RESULTSThere were 38 male and 42 female patients with a median age of 54 years. The major symptoms were upper alimentary tract hemorrhage and abdominal pain. Thirty-nine patients received local tumor excision, 18 patients underwent segmental duodenectomy, 23 patients were subjected to pancreaticoduodenectomy, all these operations were R0 resection. Thirty patients received imatinib treatment after operation, and 11 among them had metastasis relapse. Recurrence-free survival rates of 1-, 3-, and 5-years were 96.2%, 90.6%and 78.6% retrospectively. Overall survival rates of 1-, 3-, and 5-years were 100%, 98.3% and 96.1%. Multivariate Cox analysis showed tumor size >5 cm, mitotic count >5 mitosis/50 HPF and intermediate/high NIH risk classification were associated with an increased risk of recurrence. The significant difference was not detected between the limited resection group and pancreaticoduodenectomy group in OS and RFS.

CONCLUSIONSSurgery is still the main treatment for duodenum GIST. The surgical program is mainly determined by the location and size of tumor. Imatinib therapy should be used if necessary.

Abdominal Pain ; Benzamides ; Duodenal Neoplasms ; Female ; Gastrointestinal Hemorrhage ; Gastrointestinal Stromal Tumors ; Humans ; Imatinib Mesylate ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Recurrence, Local ; Pancreaticoduodenectomy ; Piperazines ; Prognosis ; Pyrimidines ; Retrospective Studies ; Survival Rate

OBJECTIVETo investigate the clinicopathological features of small gastrointestinal stromal tumors(GISTs) and to evaluate the efficacy of endoscopic therapy for GISTs.

METHODSClinicopathological and follow-up data of 418 patients with GISTs undergoing endoscopic therapy in the Zhongshan Hospital between January 2009 and July 2014 were analyzed retrospectively. All the cases were evaluated by the NIH risk classification and AIFP classification, and were grouped according to the tumor size and location. Nuclear atypia and mitotic count were used to evaluate the biological behavior of small GIST. Efficacy of endoscopic therapy was analyzed with follow-up data.

RESULTSOut of 418 patients, GISTs located in the esophagus was 14(3.3%), in the stomach 389(93.1%), in the duodenum 5(1.2%), in the rectum 10(2.4%). A total of 412(98.6%) patients were mainly spindle cells, and mitosis was not found in 320(76.5%) patients. In 389 small stomach GIST, 245(58.6%) were in fundic region. Cases were divided into four groups according to the size and the result revealed the bigger the size, the more the mitotic count. Nuclear atypia in the 1.5-1.9 cm group was significantly higher compared to other groups. Cases were divided into four groups according to the location and the result revealed the mitotic count was not associated to the location. While the nuclear atypia of stomach GIST was significantly higher than that of esophageal GIST and the nuclear atypia of rectum GIST was significantly higher than that of other positions. The median follow-up was 32(4-69) months. One case(gastric fundus GIST, >1.5 cm) presented local recurrence 23 months after operation and underwent endoscopic resection again. No recurrence or metastasis was found in other patients.

CONCLUSIONSEndoscopic resection technique is effective for small GISTs patients. The small GISTs with 0.4 cm diameter or less are often benign and should be followed up for long time. The small GISTs with 0.5 cm diameter or more possess the risk of malignancy, then surgical resection should be performed. Rectum small GISTs (except for 0.4 cm diameter or less) have worse biological behavior and should be removed.

Gastrointestinal Neoplasms ; Gastrointestinal Stromal Tumors ; Humans ; Neoplasm Recurrence, Local ; Retrospective Studies

OBJECTIVETo explore the risk factors associated with the prognosis in poorly differentiated(G3) advanced gastric cancer (AGC) and the effect of number of harvested lymph nodes on the prognosis.

METHODSClinical data of 484 patients with G3 advanced gastric cancer undergoing gastrectomy combined with lymphadenectomy in Zhongshan Hospital from December 2002 to October 2007 were retrospectively analyzed. The χ(2) test or Fisher's exact probability method was used to test measurement data. Survival was calculated using Kaplan-Meier method. Clinicopathological factors such as age, invasion depth, number of harvested lymph nodes, lymph node metastasis, ratio of metastatic lymph node, lymphatic vessels involvement were analyzed using the Cox regression model.

RESULTSNumber of lymph node dissected, lymph node metastasis, ratio of metastatic lymph node and lymphatic vessels involvement were significantly affected by groups of 15 lymph node dissected in G3 AGC patients (P<0.05). Invasion depth, number of lymph node dissected, lymph node metastasis, ratio of metastatic lymph node and lymphatic vessels involvement were significantly independent predictors of survival in G3 AGC patients (all P<0.05). Patients with at least 20 lymph nodes had a better survival rate than those with less than 20(P<0.01).

CONCLUSIONSInvasion depth, number of harvested lymph nodes, lymph node metastasis, ratio of metastatic lymph node and lymphatic vessels involvement are important factors influencing the prognosis of G3 AGC. At least 20 lymph nodes should be recommended to ensure the quality of lymphadenectomy for gastric cancer.

Gastrectomy ; Humans ; Lymph Node Excision ; Lymph Nodes ; Lymphatic Metastasis ; Prognosis ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; pathology ; surgery ; Survival Rate

Objective To prepare a nano drug(PFOB@Lip-MMC)with liposome as the carrier,liquid perfluorooc-tyl bromide(PFOB)as core and mitomycin C(MMC)loading on the liposome shell and study its inhibitory effect on the proliferation of human pterygium fibroblasts(HPFs).Methods The thin film dispersion-hydration ultrasonic method was used to prepare PFOB@Lip-MMC and detect its physical and chemical properties.Cell Counting Kit-8,Cam-PI cell viability staining and flow cytometry were employed to detect the impact of different concentrations of PFOB@Lip-MMC on the via-bility of HPFs.DiI fluorescence labeled PFOB@Lip-MMC was used to observe the permeability of the nano drug to HPFs under a laser confocal microscope.After establishing HPF inflammatory cell models,they were divided into the control group(with sterile phosphate-buffered saline solution added),PFOB@Lip group(with PFOB@Lip added),MMC group(with MMC added),PFOB@Lip-MMC group(with PFOB@Lip-MMC added)and normal group(with fresh culture medi-um added)according to the experimental requirements.After co-incubation for 24 h,flow cytometer was used to detect the apoptosis rate of inflammatory cells,and the gene expression levels of interleukin(IL)-1β,prostaglandin E2(PGE2),tumor necrosis factor(TNF)-α and vascular endothelial growth factor(VEGF)in cells were analyzed by PCR.Results The average particle size and Zeta potential of PFOB@Lip-MMC were(103.45±2.17)nm and(27.34±1.03)mV,respec-tively,and its entrapped efficiency and drug loading rate were(72.85±3.28)％and(34.27±2.04)％,respectively.The sustained-release MMC of drug-loaded nanospheres reached(78.34±2.92)％in vitro in a 24-hour ocular surface environ-ment.The biological safety of PFOB@Lip-MMC significantly improved compared to MMC.In terms of the DiI fluorescence labeled PFOB@Lip-MMC,after co-incubation with inflammatory HPFs for 2 h,DiI fluorescence labeling was diffusely dis-tributed in the cytoplasm of inflammatory HPFs.The apoptosis rate of inflammatory HPFs in the PFOB@Lip-MMC group[(77.23±4.93)％]was significantly higher than that in the MMC group[(51.62±3.28)％].The PCR examination results showed that the gene transcription levels of IL-1 β,PGE2,TNF-α and VEGF in other groups were significantly reduced com-pared to the control group and PFOB@Lip group,with the most significant decrease in the PFOB@Lip-MMC group(all P＜0.05).Conclusion In this study,a novel nano drug(PFOB@LIP-MMC)that inhibited the proliferation of HPFs was successfully synthesized,and its cytotoxicity was significantly reduced compared to the original drugs.It has good bio-compatibility and anti-inflammatory effects,providing a new treatment approach for reducing the recurrence rate after pte-rygium surgery.

OBJECTIVETo investigate the efficacy of targeted therapy combined with surgery in the treatment of recurrent and metastatic gastrointestinal stromal tumor(GIST).

METHODSClinicopathological and followed-up data of 318 patients with recurrent and metastatic GIST admitted in Zhongshan Hospital between January 2000 and December 2015 were analyzed retrospectively. According to different treatment methods, the patients were divided into four groups: surgery group (operation alone, 44 cases), target therapy group (imatinib alone, 108 cases), target therapy combined with surgery group (imatinib plus operation, 139 cases), other therapy group (chemotherapy, Chinese medicine and others, 27 cases). The progression-free survival (PFS) and overall survival (OS) of four groups were compared.

RESULTSThe baseline informations, such as age, gender, primary site, et al, were not significantly different (all P>0.05), but the recurrent and metastatic site was significantly different among 4 groups (P=0.000). The medial PFS of surgery group, target therapy group, target therapy combined with surgery was 16(95%CI: 4.9 to 27.0) months, 44 (95%CI: 30.9 to 57.1) months, 35 (95%CI: 26.5 to 43.5) months, respectively, and the latter 2 groups had significantly longer PFS than surgery group(P=0.000), while no significant difference was found between target therapy group and target combined with surgery group (P=0.251). The median OS of surgery group, target therapy group, target therapy combined with surgery, and other therapy group was 24 (95%CI: 9.0 to 39.0) months, 69(95%CI: 40.8 to 97.2) months, 92(95%CI: 78.0 to 106.0) months, 12(95%CI: 9.5 to 14.5) months. Target therapy group and target therapy combined with surgery group had significantly longer OS than surgery and other therapy groups (P=0.000), while the target therapy combined with surgery group had significantly longer OS than target therapy group(P=0.028).

CONCLUSIONTarget therapy combined with surgery can prolong the survival of recurrent and metastatic GIST patients.

Antineoplastic Agents ; therapeutic use ; Benzamides ; Combined Modality Therapy ; Disease-Free Survival ; Female ; Gastrointestinal Stromal Tumors ; drug therapy ; pathology ; surgery ; Humans ; Imatinib Mesylate ; therapeutic use ; Male ; Middle Aged ; Piperazines ; Pyrimidines ; Retrospective Studies

Objective    To explore the diagnostic value of circulating tumor cells (CTC) measured by magnetic nanoparticle method in lung cancer. Methods    (1) We measured binding capability of A549 or NCI-H1965 cell lines with recognition peptide and capture efficiency by adding tumor cells into the whole blood of healthy human. (2) We measured CTC of 34 patients suspected with lung cancer, and the counting results of CTC were compared with the following pathological results. Results    (1) The binding capability was 80.0%±6.0% for A549 and 70.1%±4.8% for H1957, while the capture efficiency was 57.3%±7.0% for A549 and 37.3%±6.1% for H1975. (2) CTCs were identified in 71.9% of patients with lung cancer. The specificity was 83.3%, and area under receiver operating characteristic (ROC) curve was 0.792 (P=0.003). Conclusion    CTC measured by magnetic nanoparticle method has promising application in the diagnosis of lung cancer.