Objective The objective of this study was to investigate the effects of maytansine on proliferation,migration,in-vasion,apoptosis and autophagy of human thyroid cancer C643 cells.Methods C643 cells were treated with different concentrations(0.049,0.195,0.781,3.125,12.5,50 and 200 μmol/L)of maytansine,the effect of maytansine on the proliferation of C643 cells was detected by the sulforhodamine B(SRB)method,and the concentration of subsequent experiments was determined.C643 cells in the logarithmic growth stage period were divided into the control group,low-dose group,mid-dose group and high-dose group.The effects of maytansine on migration and invasion abilities of C643 cells were detected by cell scratch and Transwell chamber assay;The levels of reactive oxygen species(ROS)were detected by 2′,7′-dichlorofluorescein diacetate(DCFH-DA)fluorescent probe experi-ment;The apoptosis rate of C643 cells was detected by flow cytometry;The expression of proteins related to apoptosis or autophagy was detected by Western blot.Results Maytansine at concentrations of 0.049,0.195,0.781,3.125,12.5,50 and 200 μmol/L could in-hibit the proliferation of C643 cells(P<0.05),and exhibited a significant concentration time dependence.The half maximal inhibitory concentrations(IC50)at 24,48 and 72 h were 54.255,5.193 and 0.647 μmol/L,respectively;The cell scratch and Transwell chamber results showed that maytansine at concentrations of 0.1,1 and 10 μmol/L could reduce the migration and invasion abilities of C643 cells(P<0.05 and P<0.01).The fluorescence probe results showed that maytansine at concentrations of 0.1,1 and 10μmol/L could increase the intracellular ROS levels of C643 cells(P<0.01).The flow cytometry results showed that maytansine at concentrations of 0.1,1 and 10 μmol/L could concentration dependently increase the apoptosis rate of C643 cells(P<0.01).The Western blot results showed that with the increase of maytansine concentrations,the expression of Bax protein related to apoptosis in C643 cells increased(P<0.05),the expression of Bcl-2 decreased(P<0.05),the expression of LC3Ⅱ/Ⅰ(P<0.05)and Beclin-1(P<0.01)increased,while the expression of p62 decreased(P<0.001).Conclusion Maytansine can inhibit the proliferation,migration and invasion of human thyroid cancer C643 cells,and induce the synergistic effect on apoptosis and autophagy by increasing intracellular ROS levels.

Objective The objective of this study was to investigate the effects of maytansine on proliferation,migration,in-vasion,apoptosis and autophagy of human thyroid cancer C643 cells.Methods C643 cells were treated with different concentrations(0.049,0.195,0.781,3.125,12.5,50 and 200 μmol/L)of maytansine,the effect of maytansine on the proliferation of C643 cells was detected by the sulforhodamine B(SRB)method,and the concentration of subsequent experiments was determined.C643 cells in the logarithmic growth stage period were divided into the control group,low-dose group,mid-dose group and high-dose group.The effects of maytansine on migration and invasion abilities of C643 cells were detected by cell scratch and Transwell chamber assay;The levels of reactive oxygen species(ROS)were detected by 2′,7′-dichlorofluorescein diacetate(DCFH-DA)fluorescent probe experi-ment;The apoptosis rate of C643 cells was detected by flow cytometry;The expression of proteins related to apoptosis or autophagy was detected by Western blot.Results Maytansine at concentrations of 0.049,0.195,0.781,3.125,12.5,50 and 200 μmol/L could in-hibit the proliferation of C643 cells(P<0.05),and exhibited a significant concentration time dependence.The half maximal inhibitory concentrations(IC50)at 24,48 and 72 h were 54.255,5.193 and 0.647 μmol/L,respectively;The cell scratch and Transwell chamber results showed that maytansine at concentrations of 0.1,1 and 10 μmol/L could reduce the migration and invasion abilities of C643 cells(P<0.05 and P<0.01).The fluorescence probe results showed that maytansine at concentrations of 0.1,1 and 10μmol/L could increase the intracellular ROS levels of C643 cells(P<0.01).The flow cytometry results showed that maytansine at concentrations of 0.1,1 and 10 μmol/L could concentration dependently increase the apoptosis rate of C643 cells(P<0.01).The Western blot results showed that with the increase of maytansine concentrations,the expression of Bax protein related to apoptosis in C643 cells increased(P<0.05),the expression of Bcl-2 decreased(P<0.05),the expression of LC3Ⅱ/Ⅰ(P<0.05)and Beclin-1(P<0.01)increased,while the expression of p62 decreased(P<0.001).Conclusion Maytansine can inhibit the proliferation,migration and invasion of human thyroid cancer C643 cells,and induce the synergistic effect on apoptosis and autophagy by increasing intracellular ROS levels.

Objective:To monitor and evaluate in vivo dose changes of intensity-modulated radiotherapy (IMRT) in patients with cervical cancer in a real-time manner. Methods:Twelve patients with cervical cancer admitted to our hospital were enrolled in this study. The in vivo doses were monitored by PerFRACTION?. Electronic portal imaging device (EPID) were collected in each treatment fraction for two-dimensional in vivo dose verification[γ index and dose difference (DD) index]. Log files were recorded for three-dimensional in vivo dose verification (γ index). The correlation between in vivo dose and treatment duration was analyzed by Pearson correlation analysis. Results:A total of 206 sets of EPID images and corresponding Log files were collected. The three-dimensional in vivo dose verification γ 1%/1mm of all patients was not correlated with treatment fraction ( P>0.05). Among them, the absolute difference of γ 1%/1mm of 94.66% fractions was< 1%. The mean DD 3% of two-dimensional in vivo dose verification of all patients was negatively correlated with treatment fraction ( P<0.05). Among which, the average γ 3%/3mm of 9 patients was>89% in the treatment fractions, and the average γ 3%/3mm of 98.57% fractions of these 9 patients was>93%. The other 3 patients had an average γ 3%/3mm ranged from 38% to 100%. CBCT images showed that the bladder volume of these 3 patients was significantly decreased with the relative changes by 82.08%, 84.41% and 73.59%, respectively, and the target area was retracted significantly with the relative changes by 38.12%, 59.79% and 24.46%, respectively. Conclusion:Combined with γ index and DD index, PerFRACTION? can monitor the mechanical stability of accelerator and MU delivery accuracy during treatment fractions, and monitor the changes of in vivo dose in patients with cervical cancer, which can improve the safety and quality assurance of IMRT for cervical cancer patients and provide guidance for patients with adaptive radiotherapy.

Objective To compare the immunopotentiating effects of polysaccharides extracted from wild/cultivate Cistanehe deserticola (WCDPS/CCDPS) in Xinjiang. Methods ICR mice were subcu-taneously injected twice with different doses(low,medium and high) of WCDPS and CCDPS in combination with ovalbumin (OVA). OVA-specific antibody IgG,as well as IgG1 and IgG2a subtypes, was determined by ELISA. OVA-specific lymphocyte proliferation was measured by MTT. Expression of CD4+T and CD8+T cells was analyzed by flow cytometry. Results Both WCDPS and CCDPS could significantly improve the production of OVA-specific IgG,IgG1 and IgG2a,promote the proliferation of OVA-specific lymphocytes and increase the expression of CD4+T and CD8+T cells(all P<0.05) with no significant difference between them at the same dosages (P>0.05). WCDPS and CCDPS had no influence on the body weight of mice after im-munization. Conclusion WCDSP and CCDPS could significantly enhance the OVA-specific humoral and cellular immune responses with no statistical difference and are characterized by high safety.

Objective To explore the impact of family education methods and parental rearing or not on the psychological problem tendency of primary school students.Methods The mental health test (MHT) was used on getting mental health data of the 2 838 primary school students from Dongguan city.Results (1) The results of MHT showed that 94.28％ of primary school students had no psychological problems,the tendency rate of psychological problems was 4.29％,and 1.43％ of them were suffering from serious psychological problems.Learning anxiety,physical symptoms and allergy tendency were more prominent.(2) The total MHT scores of primary school students with authoritarian,democratic,permissive and neglected family education methods were (34.84±14.58),(31.04±13.60),(35.19±12.82) and (41.19±13.10)respectively.There was no statistically significant difference in the terrorist tendency dimension scores of primary school students with different education methods (F=2.33,P=0.054),and the differences in the MHT total score and other dimensions were statistically significant (F=4.35-16.88,P＜0.01).(3) There were statistically significant differences in the scores of the total score of psychological problem tendency,dimensions of learning anxiety,anxiety to people,allergy tendency and impulse tendency of primary school students who were parental rearing or not (t=2.09-3.67,P＜0.05).However,there was no statistically significant difference in the dimensions of loneliness tendency,self-accusation tendency,physical symptoms and terrorist tendency (P＞0.05).Conclusion Family education methods and parenting styles has a impact on the mental health of primary school students.

Mitochondrial diseases are maternally inherited heterogeneous disorders that are primarily caused by mitochondrial DNA (mtDNA) mutations. Depending on the ratio of mutant to wild-type mtDNA, known as heteroplasmy, mitochondrial defects can result in a wide spectrum of clinical manifestations. Mitochondria-targeted endonucleases provide an alternative avenue for treating mitochondrial disorders via targeted destruction of the mutant mtDNA and induction of heteroplasmic shifting. Here, we generated mitochondrial disease patient-specific induced pluripotent stem cells (MiPSCs) that harbored a high proportion of m.3243A>G mtDNA mutations and caused mitochondrial encephalomyopathy and stroke-like episodes (MELAS). We engineered mitochondrial-targeted transcription activator-like effector nucleases (mitoTALENs) and successfully eliminated the m.3243A>G mutation in MiPSCs. Off-target mutagenesis was not detected in the targeted MiPSC clones. Utilizing a dual fluorescence iPSC reporter cell line expressing a 3243G mutant mtDNA sequence in the nuclear genome, mitoTALENs displayed a significantly limited ability to target the nuclear genome compared with nuclear-localized TALENs. Moreover, genetically rescued MiPSCs displayed normal mitochondrial respiration and energy production. Moreover, neuronal progenitor cells differentiated from the rescued MiPSCs also demonstrated normal metabolic profiles. Furthermore, we successfully achieved reduction in the human m.3243A>G mtDNA mutation in porcine oocytes via injection of mitoTALEN mRNA. Our study shows the great potential for using mitoTALENs for specific targeting of mutant mtDNA both in iPSCs and mammalian oocytes, which not only provides a new avenue for studying mitochondrial biology and disease but also suggests a potential therapeutic approach for the treatment of mitochondrial disease, as well as the prevention of germline transmission of mutant mtDNA.
Animals ; DNA, Mitochondrial ; genetics ; Humans ; Induced Pluripotent Stem Cells ; cytology ; metabolism ; MELAS Syndrome ; genetics ; Male ; Mice ; Microsatellite Repeats ; genetics ; Mitochondria ; genetics ; metabolism ; Mutation ; genetics

Objective To explore the clinical effect of psychological nursing pathway for patients with hepatocellular carcinoma (HCC) who are receiving 125I particle implantation therapy.Methods A total of 130 HCC patients,who were scheduled to receive 125I particle implantation therapy,were randomly and equally divided into the study group and the control group with 65 patients in each group.Traditional routine psychological nursing method was conducted for patients of the control group,while routine psychological nursing that was based on psychological nursing pathway was carried out for patients of the study group.The patient's conditions of both groups were evaluated by using self-rating anxiety scale (SAS),trait coping style questionnaire (TCSQ) and the quality of life assessment scale (SF-36).The results were statistically analyzed with t test.Results After nursing intervention,SAS scores of the study group and the control group were (43.76±6.25) and (46.16±7.19) points respectively,the difference between the two groups was statistically significant (t=-3.92,P＜0.001).The PC and NC values of the TCSQ score in the study group were (41.61±4.82) points and (20.53±5.88) points respectively,while those in the control group were (37.32±4.62) and (22.80±6.93) respectively,the differences between the two groups were statistically significant (t=5.944 and t=-3.316 respectively,P＜0.01).One month after discharge,the mental health scores of the study group and the control group were (74.58±8.68) points and (71.53±8.16) points respectively,the emotion function scores were (79.46±10.05) points and (75.13±15.09) points respectively,and the differences in both mental health score and emotion function score between the two groups were statistically significant (P＜0.05).Conclusion Standardized psychological nursing pathway can make nurses more clear about the content of psychological nursing work and enable nurses to actively plan and complete psychological nursing care measures,so that patients can get prospective,individual and comprehensive psychological care to alleviate his or her negative emotion,and to improve the quality of life.

Objective To explore the related factors influencing the prognosis of children with Tourette syndrome ( TS) . Methods We collected 420 outpatient cases of TS children in the department of pediatrics of Guangdong Provincial Hospital of Traditional Chinese Medicine from January of 2007 to October of 2010 as the research objects. Using the unified survey questionnaire, we observed the influencing factors of TS prognosis such as sex, onset age, mother’s pregnancy situation, the perinatal period, partiality for food intake, family relationship, the first symptom, the severity of disease, underlying diseases, comorbidities ( such as attention deficit hyperactivity disorder, anxiety disorder, and obsessive-compulsive disorder) , family history of psychiatric or neurological diseases, trace elements levels, electroencephalogram (EEG), antistreptolycin O (ASO), course of disease, and maintenance treatment. The related factors of prognosis was analyzed with single factor and multiple factors Logistic regression analysis. Results Among the 420 cases of TS children, 396 cases were included into the final analysis, 24 were lost and the follow-up lost rate was 5.7%. The remission rate of TS was 78.3%, and the uncured rate was 21.7%. The results of preliminary screening of the influencing factors by single factor Logistic analysis showed that the related influencing factors for TS prognosis were 12, and they were course of diseases, abnormal birth history, father’s education level, mother’s education level, upbringing methods, family history of psychiatric or neurological diseases, underlying disease history, comorbidities history, abnormal ASO, the severity of disease, the frequency of disease relapse, and the medication history of western medicine (P<0.05) . And then the obtained 12 factors were analyzed by the multiple fac tors Lo gistic regression analysis, the results showed that upbringing methods, comorbidities history, the severity of disease, and the frequency of disease relapse were correlated with TS prognosis ( P < 0.05) . Conclusion TS children will have poor prognosis when their parents spoil, indiscipline, beat and scold, and dictate them, or when the children have severe illness state, frequent recurrence of the disease, and the history of comorbidities ( atten tion deficit hyperactivity disorder, anxiety disorder, obsessive-compulsive disorder, etc.) .

Objective:To observe inhibitory effrects of DNA vaccine co-expressing CEA tandem repeat epitopes and FL on cancer cells in mice.Methods:The encoding sequences for CEA tandem repeats and FL were inserted into plasmid pcDNA3.0 using gene recombinant technique.BALB/c mice were immunized intramuscularly with the co-expressing DNA vaccine.The survival time and tumor size were measured and specific CTL cytotoxicity was detected by ~(125)I-UdR release method.Results:Compared with that of the control,the survival time was prolonged (P＜0.01)and the tumors were significantly inhibited in the mice immunized with the vaccine peDNA-triCEA_(625-667)-sFL(P＜0.01).The splenic cells from mice immunized with the vaccine pcDNA-triCEA_(625-667)-sFL induced strongly cytotoxicity against tumor cells H22-CEA ~+(P＜0.01).Conclusion:The recombinant DNA vaccine co-expressing pcDNA-triCEA_(625-667)-sFL can suppress the growth of tumor expressing CEA in mice and enhance CTL response against CEA antigen.

Objective To probe the relationship between the expression of TL1A and the level of IFN-γ secreted by T cells in the acute stage of Guillain-Barre syndrome (GBS). Methods ① Six-week female Bal b/c mice were immunized by purified recombinant human soluble TNF-like molecular 1A (rhsTL1A) protein. The polyclonal antibody against rhsTL1A was identified by immunofluorescence using human umbilical vein epithelial cells (HUVEC). ② To detect the biologic activity of rhsTL1A, the peripheral blood mononuclear cells (PBMC) from the healthy donors were separated by Ficoll gradient centrifugation and were seeded on 96-well plates with medium containing 2 μg/ml PHA (control group), 2 μg/ml PHA + 25 ng/ml rhsTL1 A, 2 μg/ml PHA + 100 ng/ml rhsTL1A and 2 μg/ml PHA + 400 ng/ml rhsTLlA respectively. T cell proliferation assay was carried out using ~3H-TdR. ③ IFN-γ productions in the sera of the children with GBS in the acute stage were detected by ELISA. ④ The ratio of CD_3~+ TL1A~+ T cells to CD_3~+ T cells in the peripheral blood of the children with GBS in acute stage was detected with flow, cytometry. ⑤PBMC from the children in acute GBS were separated and cultured in the environment adding 2 μg/ml PHA and 400 ng/ml rhsTL1A in vitro. Then, the IFN-γ in the supernatant was determined by ELISA kit after 72 hours. Results ① hTL1A A expressed by eukaryotic HUVECs was recognized by rhsTL1 A polyclonal antiserum. ② The result of T cell proliferation assay showed that SI of 25 ng/ml rhTL1A, 100 ng/ml rhTL1A A and 400 ng/ml rhTL1A group was increased compared with control group. The SI of 2 μg/ml PHA +400 ng/ml rhsTL1 A group was the highest (2. 65) among them. ③ IFN-γ productions in the sera of the children with GBS in the acute stage ((102. 25±22. 17) pg/ml) were increased significantly compared with healthy control ((28.75 ± 1.31) pg/ml, t = 3. 309, P < 0. 05). ④ The ratio of CD_3~+ TL1A~+ T cells to CD_3~+ T cells in the peripheral blood of the children with GBS in acute stage (18.22%± 1.83%) was enhanced significantly compared with healthy control (5. 17% ±0. 48%, t = 6. 884, P < 0. 01). ⑤ PBMC both in healthy control and the acute GBS secreted more IFN-γ markedly ((43.56± 4.41) pg/ml and (180.64 ± 38.39) pg/ml) after being incubated in 2 μg/ml PHA and 400 ng/ml rhsTL1A (t =4. 523 and 2. 600, P <0. 01 and 0. 05 respectively). Moreover, PBMC in acute GBS secreted more IFN-γ, than that of the healthy group markedly (t = 3. 545, P < 0. 05). Conclusions ① The mouse antiserum recognizing rhsTL1A is successfully obtained. ② In this study, 400 ng/ml rhsTL1A promotes the proliferation of T cells activated by 2 μg/ml PHA, indicating that rhsTL1A has biological activity. ③ The expression of hTL1A of activated T cells in the peripheral blood of the children with acute GBS is up-regulated. These TL1A proteins promote the secretion of IFN-γ through binding to their receptors DR_3.