Objective: To observe the protection of Testudinis Carapax et Plastrum extracts (TCPE) on serum starvation-induced PC12 cell apoptosis and explore its mechanism. Methods: The PC12 apoptosis model was established by serum starvation for 3 d. The cells were randomly divided into four groups: control group, model group, low-dose and high-dose (3 and 30 μg/mL) TCPE groups. In the three days of the treatment, cell absorbance was determined by MTT, ratio of cell apoptosis was examined by Annexin V/PI double stain flow cytometry (FCM), Caspase-3, BMP4, BMPR-IA, and p-Smad1/5/8 signaling molecular expression were detected by Western blotting, and the anti-apoptotic effect of TCPE was observed after blocking BMPs signal pathway. Semi-quantitative analysis of bands was carried out by Bio-Rad Quantity One gel analysis system. Results: MTT and FCM analyses demonstrated that TCPE could increase PC12 cell viability and decrease their apoptotic ratios in a dose dependent manner. Western blotting results showed that TCPE could decrease Caspase-3 expression, promote the expression of BMP4, BMPR-IA, and p-Smad1/5/8. There was statistically significant difference between TCPE (3 and 30 μg/mL) groups and model group (P<0.05, P<0.01) in all above results. While TCPE had no effect on the expression of BMP2, BMP7, and BMPR-II. BMPR-IB hadn't been detected. The anti-apoptotic activity was partially mitigated by neutralizing BMP4 antibody. Conclusion: TCPE has the capacity to inhibit the apoptosis of PC12 induced by serum starvation in a dose dependent manner and its mechanism may be associated with partially activating and up-regulating the expression of BMP4 signaling pathway.

Objective To determine the mechanism of nitric oxide synthase(NOS) and prostacyclin(PGI2) acting on splanchnic hyperdynamic circulation of portal hypertention(PHT). Methods Ninety-six Sprague-Dawley rats were divided into three groups, namely, intrahepatic portal hypertension(IHPH, n=31), prehepatic portal hypertension(PHPH, n=33) and sham-operated controls(SO, n=32). Animals of each group were received indomethacin(INDO) either on a short term or long term with saline as control. Portal venous pressure, together with the concentration of nitric oxide (NO) and PGI2 in serum was measured. The constitutive nitric oxide synthase(cNOS)and inducible nitric oxide synthase(iNOS)activity in the abdominal aorta and small intestine of these rats were detrmined by spectrophotometry method. RT-PCR was performed to measure the levels of iNOS and cNOS mRNA in the arteries and guts mentioned above. Results Although INDO decreased the concentration of PGI2 in serum, the long-term INDO-treated group restored splanchnic hyperdynamic circulation in both IHPH and PHPH rats, concomitant with enhanced expression of iNOS and concentration of NO(P0.05). Conclusion Overproduction of NO inducing hemodynamic abnormalities of PHT is synthesized principally by increase of iNOS. There may be a possible interaction between PGI2 and NO in hyperhemodynamics of PHT, while PGI2 may not be a mediator in the formation and development of hyperdynamic circulatory state.

Objective To develop a smoke inhalation unit simulating airtight cabin.Methods We designed a completed smoke inhalation unit composed of smoke generation cabinet,circulation pipe line and inhalation cabinet.The unit was verified with 42 SD rats inhaled with smoke generated from combustion of 9 nonmetal materials used in a airtight cabin.The rats were randomly divided into experimental group and 5 inhalation groups,with 7 rats in each group.The concentrations of CO,O2 and acid gases in the inhalation cabinet were analyzed.The activities and mortality of the rats within 7 d were recorded.COHb% of 21 rats in ten-minute inhalation groups was detected quickly after exposure.Results The concentration of smoke increased with the time of combustion and kept constant on each time point.The degree of intoxication in rats increased with the time of inhalation,and COHb% of ten-minute inhalation groups showed good reproducibility.Conclusion Our developed unit can simulate the smoke generation and intoxication in airtight cabin and keep good reproducibility of animal injury.

ObjectiveTo investigate rational surgical approaches for Siewert type Ⅰ adenocarcinoma of the esophagogastric junction (AEG),and analyze the prognostic factors.MethodsThe clinical data of 103 patients with Siewert type Ⅰ AEG who were admitted to the Renji Hospital from January 2005 to December 2009 were retrospectively analyzed.All patients were divided into transthoracic approach group (61 patients) and thoracoabdominal approach group (42 patients).The incidences of numbers of lymph node dissected and postoperative complications of the 2 groups were compared using the chi-square test,Fisher exact probability or the t test.The survival curve was drawn by the Kaplan-Meier method and the survival was analyzed using the Log-rank test.Prognostic factors were analyzed using the one-way analysis of variance and Cox regression model.ResultsNo perioperative death was observed in the 2 groups.There were significant differences in the number of lymph node dissected and number of metastatic lymph node between the 2 groups (t =2.18,2.29,P ＜ 0.05 ).There was no significant difference in splenic injury between the 2 groups (P ＞ 0.05 ).There were no significant differences in postoperative bleeding,anastomotic fistula and stricture,esophagogastric reflux,pulmonary infection and esteomyelitis between the 2 groups (x2 =0.07,0.94,0.22,1.41,0.17,P＞0.05).Of the 103 patients,97(94.2％) were followed up.The mean postoperative survival time was 26 months.The median survival time was 26 months,and the 3-yearsurvival rate was 35.9％.The 3-year survival rates of transthoracic approach group and thoracoabdominal approach group were 32.8％ and 40.2％,with no significant difference between the 2 groups ( x2 =0.37,P ＞ 0.05).The results of univariate analysis showed that radical or palliative resection,TNM stage,lymph node metastasis stage,tumor diameter and metastasis rate,degree of radical resection were independent factors influencing the prognosis of patients with Siewert type Ⅰ AEG (x2 =21.07,26.04,22.42,6.26,32.20,20.80,P＜0.05).The results of multivariate analysis showed that degree of TNM stage,lymph node metastasis rate and radical resection were independent factors influencing the prognosis of patients ( Wald =12.01,8.75,10.03,P ＜ 0.05 ).Conclusions Thoracoabdominal approach is a reasonable selection for patients with Siewert type I AEG.Degree of TNM stage,lymph node metastasis rate and radical resection were independent risk factors influencing the prognosis of patients.

Objective To investigate the effect of basic fibroblast growth factor(bFGF)on the apoptosis of scleral cells in the posterior pole in lens-induced myopia of guinea pigs and to discuss the mechanism of bFGF in inhibiting the formation of myopia. Methods Four-week-old cleaning healthy guinea pigs were randomly divided into the control group,LIM group,LIM+PBS group, LIM+the bFGF 100 ng group,LIM+the bFGF 500 ng group,LIM+the bFGF 1 000 ng group,15 in each.Except the control group, the right eye of guinea pig in other groups wore-10 D concave lens for 7 days and then different concentration of bFGF or PBS were injected into the vitreous cavity,3 days later injected again.After 15 days of-10 D concave lens treatment,the eyeballs were removed and the apoptotic cells in the scleras were determined by electron microscopy,TUNEL technique and flow cytometry.The proliferation of scleral cells in the posterior pole were determined by Ki-67 immune histochemistry stain.Results Compared with the control group,the significant differences were detected in the right eyes of LIM group(P

Animals ; Dose-Response Relationship, Drug ; Hydroxylamine ; toxicity ; Male ; Rats ; Rats, Wistar ; Spermatozoa ; drug effects ; Testis ; cytology ; drug effects

Gastrointestinal stromal tumor (GIST) is the most common mesenchymal neoplasm of the GI tract. Aberrant activation of tyrosine kinase through mutated KIT or platelet-derived growth factor receptor (PDGFRA) is the key pathogenic factor in most cases. Tyrosine kinase inhibitors (TKI) such as imatinib and sunitinib can suppress activation of tyrosine kinase receptor and has gained wide recognition as the first-line adjuvant therapy for advanced or high-risk GIST after surgery. It has become the classic model of treatment for solid tumor with molecular targeted therapy. However, the emergence of drug-resistance limits the long-term benefit of these drugs in most patients and has been a challenging clinical concern. Many factors are related to the resistance of TKI, of which KIT/PDGFRA mutation is the most important one. Genetic amplification of KIT, loss of heterozygosity, activation of an alternative downstream signaling pathways, and drug concentration are all possible factors. Therefore, reasonable individual treatment strategy and early resistance evaluation for imatinib- and sunitinib-resistant GISTs are important to patients with drug resistance in order to improve therapeutic efficacy and quality of life.
Antineoplastic Agents ; therapeutic use ; Benzamides ; therapeutic use ; Drug Resistance, Neoplasm ; genetics ; Gastrointestinal Neoplasms ; drug therapy ; genetics ; Gastrointestinal Stromal Tumors ; drug therapy ; genetics ; Humans ; Imatinib Mesylate ; Indoles ; therapeutic use ; Mutation ; Piperazines ; therapeutic use ; Protein Kinase Inhibitors ; therapeutic use ; Proto-Oncogene Proteins c-kit ; genetics ; Pyrimidines ; therapeutic use ; Pyrroles ; therapeutic use ; Receptor, Platelet-Derived Growth Factor alpha ; genetics

OBJECTIVETo explore the molecular spectra and mechanism of human hypoxanthine guanine phosphoribosyl transferase (HPRT) gene mutation induced by ethyluitrosourea (ENU).

METHODSSingle cell cloning culture, two-way screening, multiple PCR amplification and electrophoresis technique were used.

RESULTSWith dose of ENU increasing, cell plating efficiency reduced (in the group with 100-200 micro g/ml doses, P < 0.01) and mutation frequency increased (in the group with 12.5-200.0 micro g/ml doses, P < 0.05) significantly. There was no all exons deletion in spontaneous mutations, and only 7.7% of them were detected as single exon deletion. But, deletion was found in 79.7% of ENU-induced mutations (62.5%-89.4%, P < 0.01), and deletion mutations in all nine exons of HPRT gene. Most of ENU-induced mutations were chain deletion with multiple exons (88.1%).

CONCLUSIONSThe spectra in spontaneous mutations differed completely from ENU-induced ones. ENU was liable to cause bigger changes in genetic structure, which suggested a stronger ENU's mutagenesis.

Alkylating Agents ; pharmacology ; Ethylnitrosourea ; pharmacology ; HL-60 Cells ; Humans ; Hypoxanthine Phosphoribosyltransferase ; genetics ; metabolism ; Leukemia, Myeloid ; pathology ; Mutation ; drug effects ; Tumor Stem Cell Assay

Objective To study the risk factors related to recurrence of gastrointestinal stromal tumor (GIST) after discontinuing postoperative adjuvant imatinib mesylate (IM) treatment.Methods We retrospectively analyzed our clinical database of 138 GIST patients who received radical resection and subsequent IM adjuvant treatment at the Renji Hospital,Shanghai Jiaotong University School of Medicine between January 2006 and January 2014.Results For the entire Multivariate analysis study group,the overall 5-year recurrent free survival (RFS) rate was 54.5％.There were two tumor characteristics which were independent prognostic factors of GIST treated by postoperative IM:Ki67 index (P =0.005),and serosal invasion (P =0.026).The accuracy of comprehensive evaluation based on the two weighted variables was better than NIH staging criteria(AUC:0.714 vs.0.631).Furthermore,two risk groups were created according to the risk model with 5-year RFS of 81.3％ and 31.1％ as low-risk and high-risk groups,respectively (P ＜0.05).Conclusions For patients with intermediate or high risk in NIH classification,if there was tumor serosal invasion,or if there was no local invasion but Ki67 index ＞ 8％,extended continuous IM adjuvant treatment should be recommended after the primary tumor was radically resected.

Objective To analyze the characteristics of toxin, the PCR-ribotyping(RT) and the multilocus sequence typing(MLST) of Clostridium difficile strains isolated from China-Japan Friendship Hospital in order to provide a basis for monitoring the outbreak of nosocomial Clostridium difficile infection.Methods A total of 321 samples were collected from the patients with suspected Clostridium difficile infection(CDI) in China-Japan Friendship Hospital(CJFH) during 2012 to 2013.All Clostridium difficile strains were isolated and identified by the standard phenotypic culture method.Cytotoxicity test was performed to detect toxin B.Toxin genes (tcdA and tcdB) and binary toxin genes (cdtA and cdtB) harbored by those strains were analyzed.RT and MLST were used for homologous analysis.Clinical data of the patients were collected to analyze the isolation rate of Clostridium difficile in different populations.Results Forty-eight strains of Clostridium difficile were isolated from 46 patients with diarrhea and three of them were isolated from the same patient.The incidence of CDI among all patients, outpatients and inpatients were 14.3%(46/321), 12.8%(5/39) and 14.5%(41/282), respectively.Toxin B was detected in all of the strains as indicated by the cytotoxicity test.Strains of sequence type 1(ST1) showed the strongest cytotoxicity of all the isolated Clostridium difficile strains.Ten out of the 48 strains (20.8%) were tcdA(-)/tcdB(+) strains, which belonged to either ST37 or ST81.The results of RT and MLST were consistent in assigning the strains into nine types, in which the predominant type was ST1/RT027 accounting for 27.1% (13/48).All of the ST1/RT027 strains presented a toxin gene profile of tcdA(+)/tcdB(+) and cdtA(+)/cdtB(+).Most of the ST1/RT027 strains were isolated from the Traditional Chinese Medicine Department of Respiratory, where smallnosocomial outbreaks of ST1/RT027 strain infection might happen.Conclusion CDI diagnosed in CJFH mainly belongs to nosocomial infection.Most of the isolated strains harbor tcdA(+)/tcdB(+) genes.Surveillance for the outbreaks of CDI caused by ST1/RT027 strains over producing toxins A and B should be strengthened in hospitals.