BACKGROUND:5-Fluorouracil is a common chemotherapy drug for gastric cancer, but it is more likely to develop drug resistance in clinical treatment. Studies have shown that tumor stem cels are lowly sensitive to chemotherapeutic drugs, which may be an important cause of chemotherapy resistance. OBJECTIVE:To analyze the sensitivity of gastric cancer stem cels to 5-fluorouracil in vitro, and to understand the mechanism of drug resistance associated with gastric cancer. METHODS:Based on the sorting strategy, human gastric cancer cel clones were isolated from AGS cel lines. CD44 and thymidylate synthetase expression in different clones was detected using immunocytochemistry analysis. Clone formation assay was used to evaluate self-renewal capacity of different clones. Cel counting kit-8 was used to determine the clonal growth inhibition rate of AGS under treatment with different concentrations of 5-fluorouracil. RESULTS AND CONCLUSION:The AGS cels that were inoculated with low density and cultured could differentiate into 32 forms, including the ful clone (16%, 5/32), the second clone (66%, 21/32) and the accessory clone (19%, 6/32). Among them, the ful clones highly expressed CD44 and thymidylate synthetase, and could generate a great amount of passage 2 clones after inoculation; the second clones showed a weak expression of CD44 and thymidylate synthetase, and could generate a smal number of passage 2 clones after inoculation; the accessory clones showed a weak or no expression of CD44 and thymidylate synthetase, and there was no passage 2 clone after inoculation. Under the effect of different concentrations of 5-fluorouracil, the growth inhibition rates of the secondary clones and AGS cels were both higher than that of the ful clones (bothP < 0.05). These findings indicate that gastric cancer stem cels have a relatively lower sensitivity to 5-fluorouracilin vitro, which is speculated to be an important mechanism of drug resistance.

Objective To investigate the method of handmade microwire lasso device for the management of coil migration during intracranial aneurysm embolization. Methods Two migration coils were removed from intracranial arteries using handmade microwire lasso rings. The first coil prolapsed completely from an aneurysm sac and flowed to the middle cerebral artery M2 segment. The second coil partially prolapsed out into a parent artery. The microwire lasso device was made of 4-0 silk thread and fixed using 0. 36 mm (0. 014 inch)microwire and 0. 43 mm (0. 017 inch)microcatheter. Results After removing 2 migration coils from intracranial arteries,angiography did not reveal vascular injury and thrombosis. There was no aneurysm rupture. After awaking from anesthesia,the patient did not have neurological deficit. The patient was followed up for 3 months after procedure. MR angiography confirmed the patency of related arteries. Conclusion The handmade microwire lasso device for the management of coil migration in the process of aneurysm embolization is a simple,effective,and economical method.

Objective To investigate the influencing factors for late-preterm birth of twins. Methods We retrospectively analyzed the clinical data of 301 twins delivered in Beijing Haidian Maternity and Child Health Hospital between August 1, 2011 and August 31, 2013. Twins with late-preterm births were classified as the observation group(n=138) and those delivered at term were classified as the control group (n=163). Comparison between the two groups was conducted on both maternal and neonatal complications. Two independent sample Student's t test, Chi-square test and logistic regression analysis were performed for statistical analysis. Results (1) The ratio of two male fetuses, dichorionic diamniotic twins, and iatrogenic preterm birth were 41.3% (57/138), 76.1% (105/138) and 65.2% (90/138), respectively, in the observation group, while 23.9% (39/163), 89.6% (146/163) and 96.9% (158/163), respectively, in the control group (χ2=10.40, 9.81 and 53.59, respectively, all P<0.05). Mean birth weight of twins in the observation group was less than that in the control group [(2 450±349) vs (2 640±304) g, t=7.12, P<0.05]. Maternal age, gravidity, parity, history of abortion, mode of conception and 1 min Apgar score of the neonates between the two groups were not significantly different (all P>0.05). (2) The risk factors for late-preterm birth of twins included hypertension diseases complicated pregnancy, gestational diabetes mellitus, premature rupture of membranes, fetal distress, anemia and umbilical cord abnormality [OR=0.190(95%CI: 0.094-0.387), 1.980 (95%CI: 1.009-3.883), 0.030(95%CI: 0.007-0.131), 0.062(95%CI: 0.006-0.642), 0.470(95%CI:0.243-0.908) and 2.779(95%CI:1.093-20.736), respectively, all P<0.01 or 0.05]. (3) The incidences of hyperbilirubinemia, respiratory diseases, hypoglycemia, anemia, and neonatal infection were 27.9%(77/276), 4.4%(12/276), 4.7%(13/276), 4.4%(12/276) and 3.6%(10/276), respectively, in the observation group, which were all higher than those in the control group [9.5%(31/326), 0.6%(2/326), 1.2%(4/326), 0.6%(2/326) and 0.3%(1/326), respectively,χ2=34.33, 9.18, 6.61, 9.18 and 9.16, respectively, all P<0.05]. The incidence of small for gestational age in the observation group was lower than that in the control group [3.6%(10/276) vs 12.3%(40/326),χ2=3.86, P<0.05]. Other complications in the two groups (intracranial hemorrhage, acidosis, and hemolysis) were not significantly different (all P>0.05). Conclusions Late-premature births in twin pregnancies are related to many maternal and fetal risk factors. Therefore, clear understanding of these risk factors might improve the pregnancy outcomes.

Objective To explore the clinical significance of oral pancreatin capsules for patients after total gastrectomy.Methods Ninety cases of total gastrectomy patients in Nanyang City Center Hospital were selected and randomly divided into trypsin group and control group,with 45 patients in each group.The control group were treated by conventional therapy,and the patients of trypsin group were treated by pancreatin capsules on the basis of conventional therapy after 3 weeks postoperation.Scores of EORTC QLQ-C30,korenaga and fecal fat contents were compared between two groups six months postoperation.Results No statistically significant difference was found in the scoring system of EORTC QLQ-C30,korenaga and fecal fat contents in two groups before administered,The score of role function,emotional function and overall health status in the scoring system of EORTC QLQ-C30 of patients in trypsin group were higher than control group after 6 months administration.The differences were statistically significant (P<0.05 ).The score of fatigue,loss of appetite,nausea,vomiting,diarrhea in the scoring system of EORTC QLQ-C30 of patients in trypsin group were lower than control group after 6 months administration.The differences were statistically significant (P<0.05).The loss of appetite,swelling postprandial satiety,insomnia and the extent of weight loss in the scoring system of korenagain trypsin group were lower than control group after 6 months administration.The differences were statistically significant (P<0.05 ).The fecal fat contentsin trypsin group were lower than control group at 24 h and 72 h.The differences were statistically significant (P<0.05).The cases of 24h fecal fat content>7g in trypsin group was lower than control group at 24 h.The differences were statistically significant (P<0.05).Conclusion Oral pancreatin capsules could significantly improve the quality of life of patients after total gastrectomy, which is worthy of promoting.

ObjectiveTo evaluate the effects of selective cyclooxygenase-2 inhibitor nimesulide lone and combined with cisplatin on tumor growth,and Ki67 and Caspase-3 expression in lung cancer xenografts in nude mice. Methods The mice were randomly divided into 4 groups:the control group,the nimesulide group,the cisplatin group and the nimesulide combined with cisplatin group. A549 cells were injected into BALB/c nude mice subcutaneously.On the 21st day after treatment tumor tissues were collected,and the xenografts growth were observed. The expression of Ki67 and Caspase-3 were detected by immunohistochemical method.ResultsNimesulide combined with cisplatin could significantly inhibited the xenografts growth compared with nimesulide or cisplatin.The tumor inhibition rate was 44.33％ in the nimesulide group,53.61％ in the cisplatin group and 80.41％ in the nimesulide combined with cisplatin group (P ＜ 0.05).Immunohistochemical analysis showed that the expression rates of Caspase-3 was significantly increased in the nimesulide combined with cisplatin group (67.43 ± 23.57 ) ％,the cisplatin group (48.40 ± 20.37 ) ％,and the nimesulide group (38.65 ± 15.37)％,compared with the control group (27.63 ± 13.03)％ (P ＜ 0.05).The expression rate of Caspase-3 was significantly increased in the nimesulide combined with cisplatin group compared with the nimesulide group or the cisplatin group (P ＜ 0.05 ).The expression rate of Ki67 was significantly decreased in the nimesulide group combined with cisplatin group (24.34 ± 15.90)％,the cisplatin group (40.85 ± 22.47)％ and the nimesulide group (53.33 ± 19.67)％ compared with the control group ( 80.43 ± 16.88 ) ％ ( P ＜ 0.05 ).The expression of Ki67 was significantly decreased in the nimesulide combined with cisplatin group compared with the nimesulide group or the cisplatin group ( P ＜ 0.05 ).Conclusion Nimesulide can inhibit human lung cancer A549 cells xenografts in nude mice growth,Nimesulide enhanced the inhibitory effects of cisplatin.The mechanism may be related to inhibition of tumor cell Ki-67 expression,increased expression of Caspase-3,inhibition of tumor cell proliferation,and inducing tumor cell apoptosis.

p21 activated kinase 4 (PAK4) is a member of a family of serine/threonine kinase.PAK4plays an important role in a variety of cellular functions including cell cycle,cell proliferation,apoptosis,and cytoskeletal reorganization.Recently,PAK4 has been shown to overexpress in a variety of malignancies,and contribute to cancer cell migration and invasion through multiple signalling pathways.

Objective To observe the changes of NK cell subset (CD56+,CD16+CD56+,CD16+),T cell subset (CD4+,CD8+,CD4+/CD8+) counts and related cytokines (IL-2,IL-4,INF-γ) in children with viral pneumonia.Methods Thirty-two children with viral pneumonia in acute stage (within 2 days after pneumonia onset) and recovery phase (within the range of the third to the fifth day after pneumonia onset) were included in this study.Peripheral blood NK cell subsets and T cell substes were determined by the flow cytometry.Blood IL-2,IL-4 and INF-γ were detected by ELISA.NK cytoactivity was measured by LDH release method.Results (1) The levels of the CD16+CD56+ and CD16+NK cell counting in acute stage [(0.73±0.17)% and (0.39±0.20)%] were lower than those in the recovery phase [(1.47±0.22)% and (0.89±0.14)%],which showed significant difference (P<0.01),however the level of CD16+CD56+ and CD16+NK cell counting either in acute stage or recovery phase was significantly lower than those of healthy control group (P<0.01).The sub population counting and NK cell activity was directly correlated.CD56+NK cell counting showed no significant difference between viral pneumonia group and control group (P>0.05).(2) There was no significant difference in blood IL-2 and IL-4 level between viral pneumonia group (either in acute stage or recovery phase) and the control group (P>0.05).As compared with that of the control group,blood INF-γ level of viral pneumonia group showed no significant change in acute stage (P>0.05),but INF-γ level in recovery phase [(28.10±1.38)?μg/L] was higher than that in acute stage [(22.78±1.19)?μg/L] and there was significant difference (P<0.01).(3) As compared with that of the control group,CD4+ and CD4+/CD8+T cell counting of viral pneumonia group showed no obvious changes either in acute stage or recovery phase (P>0.05).CD8+T cell counting of both two stages were much lower than that of the control group (P<0.05),but there was no significant difference between the two stages (P>0.05).Conclusion The NK cell activity in children with viral pneumonia decline obviously,which might be related to the changes of T cell subsets;the activity of suppressor T cell was depressed in patients with viral pneumonia.There are maybe many factors involved in the NK cell activation.

To study on one case of severe cerebral parlsy. Special education and rehibilitative training for children with severe cerebral parlsy arc investigated in the current study.

AIM:To explore the effects of the combined use of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (ASODN) and cisplatin on apoptosis of HL-60 cells. METHODS:PCR-ELISA was used to determine telomerase activity in HL-60 cells untreated or treated with ASODN, the expression levels of hTERT protein were assayed by immunofluorescence using fluoresce isothiocyanate (FITC) label. Apoptosis was detected by DNA electrophoresis and flow cytomertric cell cycle analysis. RESULTS:The expression levels of hTERT protein and telomerase activity in HL-60 cells treated with ASODN were shown to decrease with time, the DNA fragments were obviously found and the percentage of apoptosis cells was significantly enhanced in HL-60 cells with the combined use of hTERT ASODN and cisplatin compared with the combined use of hTERT sense oligodeoxynucleotide and cisplatin or cisplatin alone, respectively. CONCLUSION:Inhibition of telomerase activity by hTERT ASODN increases the cisplatin-induced apoptosis of HL-60 cells.

Objective To investigate the clinical manifestation, biochemically detected data, and radiographic features of a pedigree with suspected mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome, and to explore the correlations between the clinical features and the mutant heteroplasmy levels of mitochondrial genome. Methods The personal details, histories of stroke-like episodes and seizures within the proband and 11 members in the maternal lineage of the family were collected. Routine blood examinations and plasma lactate levels before and after movements of these family members were detected, followed by cephalic MRI examinations. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing were used to detect and validate the A3243G point mutation in mitochondrial genome, and real-time PCR were used to quantify the mutation proportion of A3243G. Results Typical symptoms of MELAS such as seizures, stroke-like episodes and hyperlactacidemia and atypical symptoms such as growth failure, exercise intolerance, fevers and migraines were observed on several members in the pedigree. Cephalic MRI findings performed during episode periods were in accord with the typical radiographic features of MELAS and cerebellar atrophy was commonly observed. Family members on the maternal side all harbored the point mutation on 3243 site in mitochondrial genome. Meanwhile, patients with higher heteroplasmy levels relatively manifested more typically and severely according to the clinical observation. Conclusions The pedigree is diagnosed with maternal inheritance of MELAS syndrome. The main cause can be attributed to a mitochonorial A3243G mutation.The mutant heteroplasmy levels of hemocytes in peripheral blood are positively associated with genetic relationship, seizure anticipation, plasma lactate data and other clinical features.