Objective:Assessing the therapeutic efficacy of methotrexate(MTX)-modified magnetic nanoparticles in thermo-chemotherapy for rat breast cancer and its impact on immune function.Methods:Female Wistar rats were subcutaneously inoculated with breast cancer Walker-256 cells to establish a transplantation tumor model,and injected with polyethyleneimine(PEI)-modified Fe3O4 magnetic nanoparticles(47T group,42T group and multiple 42T group)or MTX-modified Fe3O4 magnetic nanoparticles(47TC group,42TC group and multiple 42TC group)for thermotherapy under the magnetic field at different temperatures(47℃and 42℃).The rats injected with MTX-modified magnetic fluid only(MFC group)and the tumor-bearing rats without any treatment(blank control group),with irradiation treatment in an alternating magnetic field only for 30 minutes(M group),with injection of PEI-modified magnetic fluid only(MF group),with treatment of MTX-mono drug(MTX group)and not inoculated with tumor cells(normal group)were used as control groups.X-ray radiography was used to display the distribution of magnetic fluid in the tumor tissue 24 hours,2 weeks and 2 months after intra-tumor injection.After 24 hours of treatment,three rats were selected from each of the 47T and 47TC groups,and the effect of magnetic fluid on tumor cells was observed under an electron microscope after execution.After 14 days of treatment,the tumor volume of rats was measured and statistically analyzed.At the same time,4 rats were selected from each of the 47TC,47T,42TC,42T,MFC,MTX,blank control and normal groups,and the levels of IL-2,IFN-γ and IL-4 in peripheral blood were detected by ELISA method.The remaining rats were observed for long-term survival.Results:The magnetic nanoparticles were evenly distributed in the center of the tumor but unevenly distributed at the tumor's edge;they primarily localize amomg tumor cells and can penertrate into tumor cells.Tumor growth was inhibited in rats in the 47TC,47T,multiple 42TC and multiple 42T groups(all P<0.05),and the survival rates of the rats were high.As compared with the blank control group,the levels of IL-2 and IFN-γ were increased while the IL-4 level was decreased in the 47TC and 47T groups(all P<0.05).Conclusion:Thermo-chemotherapy at 47℃for 30 minutes and multiple sessions at 42℃for 60 minutes can partially inhibit tumor growth and prolong rat survival.This effect maybe related to the thermo-chemotherapy at 47℃for 30 minutes which can activate the body's immune function.

Objective:Assessing the therapeutic efficacy of methotrexate(MTX)-modified magnetic nanoparticles in thermo-chemotherapy for rat breast cancer and its impact on immune function.Methods:Female Wistar rats were subcutaneously inoculated with breast cancer Walker-256 cells to establish a transplantation tumor model,and injected with polyethyleneimine(PEI)-modified Fe3O4 magnetic nanoparticles(47T group,42T group and multiple 42T group)or MTX-modified Fe3O4 magnetic nanoparticles(47TC group,42TC group and multiple 42TC group)for thermotherapy under the magnetic field at different temperatures(47℃and 42℃).The rats injected with MTX-modified magnetic fluid only(MFC group)and the tumor-bearing rats without any treatment(blank control group),with irradiation treatment in an alternating magnetic field only for 30 minutes(M group),with injection of PEI-modified magnetic fluid only(MF group),with treatment of MTX-mono drug(MTX group)and not inoculated with tumor cells(normal group)were used as control groups.X-ray radiography was used to display the distribution of magnetic fluid in the tumor tissue 24 hours,2 weeks and 2 months after intra-tumor injection.After 24 hours of treatment,three rats were selected from each of the 47T and 47TC groups,and the effect of magnetic fluid on tumor cells was observed under an electron microscope after execution.After 14 days of treatment,the tumor volume of rats was measured and statistically analyzed.At the same time,4 rats were selected from each of the 47TC,47T,42TC,42T,MFC,MTX,blank control and normal groups,and the levels of IL-2,IFN-γ and IL-4 in peripheral blood were detected by ELISA method.The remaining rats were observed for long-term survival.Results:The magnetic nanoparticles were evenly distributed in the center of the tumor but unevenly distributed at the tumor's edge;they primarily localize amomg tumor cells and can penertrate into tumor cells.Tumor growth was inhibited in rats in the 47TC,47T,multiple 42TC and multiple 42T groups(all P<0.05),and the survival rates of the rats were high.As compared with the blank control group,the levels of IL-2 and IFN-γ were increased while the IL-4 level was decreased in the 47TC and 47T groups(all P<0.05).Conclusion:Thermo-chemotherapy at 47℃for 30 minutes and multiple sessions at 42℃for 60 minutes can partially inhibit tumor growth and prolong rat survival.This effect maybe related to the thermo-chemotherapy at 47℃for 30 minutes which can activate the body's immune function.

Objective:To explore the potential mechanism of PD-1 inhibitor P on RIMI from the perspective of immune microenvironment.Methods:To establish a mouse model of radiation-induced myocardial injury (RIMI), twenty C57BL/6 mice were randomly divided into 4 groups, 5 in each group. Group A was the healthy control group; Group B was the PD-1 inhibitor group; Group C was the simple irradiation group, with a heart irradiation of 15 Gy; Group D was the irradiation+ PD-1 inhibitor group. One month after irradiation, the mice were anesthetized and sacrificed. The morphological changes of myocardial tissues were observed by HE staining. The myocardial fibrosis was assessed by Masson staining. CD 3+ , CD 3+ CD 4+ , CD 3+ CD 8 lymphocyte subsets and cytokines (IL-4, IL-6, IL-17A, TNF-α, TGF-β 1 and INF-γ) levels were determined by flow cytometry. The apoptosis rate of myocardial cells was detected by TUNE. Results:One month after irradiation, there was no obvious myocardial fibrosis in group B, and collagen fibers were distributed in the interstitium of myocardial cells in groups C and D. Semi-quantitative analysis results showed that the myocardial collagen volume fraction (CVF) of groups A, B, C and D were (1.97±0.36)%, (2.83±1.03)%, (5.39±0.77)% and (7.72±1.43)%, respectively. The CVF between group A and group B was similar ( P=0.314), and the differences in CVF between the other groups were statistically significant (all P<0.05). Compared with group A, the absolute value and percentage of CD 3+ T lymphocytes were significantly increased in groups B, C and D (all P<0.01). The values in group D were significantly higher than those in group B and group C (all P<0.01); The absolute value and percentage of CD 3+ CD4 T lymphocytes were similar among four groups (all P>0.05); The absolute value and percentage of CD 3+ CD 8 T lymphocytes in group D were significantly higher than those in groups A, B and C (all P<0.001). The expression levels of IL-6, IL-17A, and TGF-β 1 in group D were significantly higher compared with those in groups A, B and C (all P<0.001). The apoptotic index was gradually increased in four groups, and the differences in apoptotic index among four groups were statistically significant (all P<0.001). Conclusion:PD-1 inhibitors can aggravate RIMI by promoting myocardial immune inflammatory response.

Objective:To explore the establishment of radiation-induced heart damage (RIDH) SD rat models caused by irradiation of 15Gy/3f and the changes in early detection indicators, and evaluate the effect of irradiation combined with recombinant human endostatin (Endostar).Methods:75 adult male SD rats were randomly divided into the blank control group (C group), Endostar group (E group), 25Gy irradiation group (MHD 25 group), 15Gy irradiation group (MHD 15 group) and 15Gy irradiation combined with Endostar group (MHD 15+ E group), respectively. Blood sample was taken to measure the CK, CK-MB, LDH and CRP at 24h, 48h and 15d after corresponding interventions. After cardiac echocardiography at 1, 3 and 6 months, 5 rats in each group were randomly sacrificed and myocardial tissues were collected for HE and Masson staining. Two-way ANOVA was employed for statistical analysis. Results:Compared with group C, myocardial fibrosis were observed in the MHD 15 group at 6 months ( P<0.05), which occurred later than that in the MHD 25 group. Ejection fraction (EF) and fractional shortening (FS) were significantly decreased after 3 months in each irradiation group (all P<0.05), whereas the degree of decrease was similar among all groups (all P>0.05). The expression levels of myocardial enzymes and inflammatory cytokines did not significantly differ among different groups (all P>0.05). Conclusions:In the early stage, exposure to 15Gy/3f irradiation can cause cardiac function damage in SD rat hearts, such as the reduction of EF and FS, and even lead to myocardial fibrosis in the late stage, which is delayed and less severe than high-dose irradiation. Irradiation combined with Endostar has no significant effect on radiation myocardial injury in rats.

Objective:To explore the possibility of CD 4+ T cells and CD 4+ /CD 8+ ratio in peripheral blood to predict the survival of patients with stage Ⅳ non-small cell lung cancer (NSCLC), and to establish a Nomogram prediction model. Methods:The influence of CD 4+ T cells and CD 4+ /CD 8+ ratio on the clinical factors and survival of 682 patients pathologically diagnosed with stage Ⅳ NSCLC with no history of cancer treatment was retrospectively analyzed and the Nomogram prediction model was established. Combined with the changes of immune cells levels in 110 patients after treatment, the prognostic and predictive values of CD 4+ T cells and CD 4+ /CD 8+ ratio were verified. Countable data were analyzed by t-test. The survival rate was calculated by Kaplan-Meier method, log-rank test or univariate analysis. The multivariate analysis was performed by Cox regression model. Results:Univariate analysis demonstrated that CD 4+ > 43.15% before treatment significantly prolonged the survival. By multivariate analysis of Cox regression model, CD 4+ >43.15% was an independent prognostic factor to prolong survival for stage Ⅳ NSCLC. The Nomogram model was established and verified that the predicted and actual overall survivals were highly consistent. Further analysis showed that 43.15% as the critical value of CD 4+ T cell level significantly prolonged survival when CD 4+ expressed at a high-level before treatment, after treatment, before and after treatment, or combined with CD 4+ /CD 8+ >1.65. Conclusions:The baseline level of CD 4+ T cells before treatment in peripheral blood is an independent prognostic factor for stage Ⅳ NSCLC. The CD 4+ /CD 8+ ratio before treatment has limited value in predicting the prognosis.

Objective:To explore the changes of CD 8+ T cells in stage Ⅲ-Ⅳ non-small cell lung cancer (NSCLC) patients before and after radiochemotherapy and evaluate its clinical value in predicting survival. Methods:A total of 795 patients with stage Ⅲ-Ⅳ NSCLC who completed CD 8+ T cell testing from January 2011 to December 2017 were recruited (249 patients completed 1-3 tests within 6 months after treatment). The survival difference of patients with different levels of CD 8+ T cells and the prognostic value of the changes in the CD 8+ T cell level were analyzed. The survival analysis was performed by Kaplan- Meier method and log-rank test or univariate analysis. The multivariate survival analysis was conducted by Cox’s regression model. Results:Before treatment, the levels of CD 8+ T cells in the peripheral blood did not significantly differ among patients with different clinical factors. The survival time of stage Ⅲ NSCLC patients with CD 8+ T cell levels of<26.44% was significantly prolonged ( P=0.043). After treatment, the levels of CD 8+ T cells were significantly higher than those before treatment. The levels were similar within 1-3 months, decreased after 4-6 months but still significantly higher than those before treatment. The median survival time of patients with CD 8+ cell levels of<43.90% after treatment was 22 months, significantly longer than 16 months of those with CD 8+ cell levels of ≥43.90%( P=0.032). Stratified analysis demonstrated no significant difference in the survival time at 1 month and 2-3 months after treatment ( P>0.05), whereas the survival time significantly differed at 4-6 months ( P=0.001). The multivariate survival analysis showed that CD 8+ cell levels of<43.90% after treatment was an independent prognostic factor ( HR=0.714, P=0.031). Conclusions:The effect of CD 8+ T cells on prognosis of patients with stage Ⅲ-Ⅳ NSCLC is limited. After treatment, CD 8+ T cell levels are increased significantly. A certain increase in the CD 8+ T cell levels can prolong the survival time. The detection of CD 8+ T cell subtypes plays a more significant role.

Objective To investigate the impact of the changes of posttreatment karnofsky performance status (KPSpost) on the overall survival (OS) for patients with stage Ⅳ non-small cell lung cancer (NSCLC) underwent concurrent chemoradiation.Methods A total of 279 patients (male 198 and female 81) with histological confirmed stage Ⅳ NSCLC were enrolled in this study with a median age of 58 years old (range 22 to 80 years old).There were 166 cases of squamous carcinoma,87 cases of adenocarcinoma,and 22 cases of unclassified carcinoma,respectively.All enrolled patients received more than 2 cycles of chemotherapy and more than 36 Gy of concurrent radiotherapy.Kaplan-Meier method and Log-rank test were applied to evaluate OS.Multivariate analyses were carried out by the Cox proportionalhazard model.Chi-square test and logistic regression analysis were used to explore the related factors of KPSpost.Results There were 198 patients with improved KPSpost and 81 patients with decreased KPSpost,respectively.Univariate and multivariate analyses indicated that the improvement of KPSpost was associated with longer OS.Logistic regression analysis showed that the improvement of KPSpost was positively related with treatment of more than 4-6 cycles chemotherapy concurrent with over 63 Gy radiation to primary tumor.The improvement of KPSpost also correlated positively with disease control rate (DCR),but negatively with PLT toxicity and radiation esophagitis.Conclusions KPSpost was an independent prognostic factor of OS for patients with stage Ⅳ NSCLC underwent concurrent chemoradiation.Chemotherapy of 4-6 cycles and concurrent over 63 Gy radiotherapy dose to primary tumor,as well as DCR were positive factors for KPSpost improvement.However,stage 3-4 PLT toxicities and radiation esophagitis decreased the KPSpost.

Objective To quantitatively compare the effect of preservation or removal of atlas posterior arch on cervical posterior decompression,so as to provide a basis for reasonable selection of upper cervical spine decompression range and determination of surgical indications for atlas posterior arch resection.Methods The data of 45 patients with posterior decompression of upper cervical spine were retrospectively analyzed.According to the decompression range,the patients were divided into C2-C7 group and C1-C7 group.There were 25 cases in the C2-C7 group,19 males and 6 females,with an average age of 56.3 years (40-71 years),4 cases of cervical spondylotic myelopathy and 21 cases of ossification of the posterior longitudinal ligament of cervical spine.All of the 25 patients underwent open-door laminoplasty:20 cases with hinge side anchoring procedure and 5 cases with preservation of the unilateral posterior muscular-ligament complex procedure (titanium cable procedure).There were 20 cases in C1-C7 group,12 males and 8 females,with an average age of 58A years (44-75 years).All of the 20 cases underwent atlas posterior arch resection as well as C2-C7 open-door laminoplasty,including anchoring procedure in 1 case,titanium miniplate procedure in 4 cases,and titanium cable procedure in 15 cases.Standardized vertebral-cord distance (SVCD) at each level from atlas to level C~ was measured on T2-weighted images of MR on the mid-sagittal plane in the neutral position pedormed 3-12 months postoperatively at each individual level.As the main outcomes,the SVCD values obtained at the same level of the two groups were compared between the two groups.Shapiro-Wilk normality test was performed on the SVCD values at C1.2 and C2 levels of two groups.The area under the normal distribution curve of SVCDs was used to calculate the corresponding residual compression rate with different magnitude of compression mass to further discover the difference of the decompression effect between the two groups.Results The SVCD obtained at the level of the anterior arch of atlas (C1),the junction of odontoid process and axis (C1,2) and the middle part of axis body (C2) in the C2-C7 group was 9.91±1.34 mm,8.35±1.27 mm,and 8.22 ±1.43 mm,respectively.The SVCD at the same levels was 11.02±1.60 mm,9.72±1.24 mm,and 9.12±1.11 mm,respectively.SVCDs differed significantly in the above range between the two groups.However,from level C2,3 to C6,7,there was no significant difference in SVCDs between the two groups.The JOA score of group C2-C7 was 11.8±2.7 preoperatively and increased significantly to 14.7±1.8 at 12 months postoperatively(t=-7.006,P＜0.001) with a recovery rate of 57.0％±32.2％.The JOA score of group C1-C7 was 11.7±2.8 preoperatively and increased significantly to 14.2±2.3 at 12 months postoperatively(t=-6.177,P＜0.001) with a recovery rate of 51.9％±32.1％.Conclusion Atlas posterior arch resection can significantly increase the decompression effect of posterior cervical surgery from the anterior arch of atlas to the middle part of axis body,but it would not increase the decompression effect at level C2.3 or below.When the magnitude of the ventral compression factor exceeds the decompression limit (8.5 mm) available with C2-C7 decompression in the range from atlas to the middle of the axis body,extending the decompression range by atlas posterior arch resection is an effective means to achieve adequate decompression.

Objective　To investigate the clinical efficacy of anterior cervical surgery by comparative analyzing hand dysfunction using brief Michigan hand questionnaire(Brief MHQ)in Hirayama disease patients.Methods　From Aug 2011 to Dec 2016,27 patients of hirayama disease who underwent surgery were enrolled in this study.The study group consisted of 27 men.The mean follow-up period was 41.1 months.The levels of surgery included 18 cases of C4-C7,6 cases of C3-C6,2 cases of C4-C6 and 1 case of C5-T1.Brief MHQ were evaluated for the 27 patients.According to the Wilcoxon analysis,the unchanged domains were analyzed with the multifactor Logistic regression analysis by preoperative duration of symptoms,age of onset,and number of affected extremities.Dynamic flexion-extension lateral X-rays were performed at baseline and at final follow-up.Results　No failure of internal fixation was detected on dynamic flexion-extension lateral X-rays.Five domains of preoperative Brief MHQ had lower scores,including Function,Satisfaction,Aesthetics,Activities of daily living,Work domain.With the exception of Aesthetics and Pain domain,all the other four domains showed significant improvement after surgery.The total score was 38.44±5.83 at base-line and 43.19±4.47 at follow-up.The score of Function was 5.19±1.36 at baseline 6.37±1.15 at follow-up;The score of Satisfaction was 5.56±1.22 at baseline 6.60±1.05 at follow-up;The score of Activities of daily living was 6.33±1.84 at baseline 7.60±1.47 at follow-up;The score of Work was 6.85±1.75 at baseline 7.67±1.33 at follow-up.The risk factors of postoperative outcomes reported in the literature included duration of disease at the time of surgery,age of onset,and extremity involvement.According to the Logistic regression,pre-operation duration was the risk factor for Aesthetics domain and the cut-off time was 1.75 years.Conclusion　Four domains of Brief MHQ score were improved significantly after anterior surgery for patients with hirayama disease.Brief MHQ was useful to evaluate the hand dysfunction and clinical efficacy in patients with hirayama disease.

Objective To evaluate the clinical efficacy and toxicity of concurrent pemetrexed-cisplatin (PP) or docetaxel-cisplatin (DP) with intensity-modulated radiation therapy (IMRT) in patients with stageⅠV lung adenocarcinoma. Methods Stage IV lung adenocarcinoma patients with unknown EGFR mutation status or wild-type admitted to Guizhou Cancer Hospital from 2011 to 2016 were randomly assigned into the PP (n=50) and DP groups (n=51).All patients received concurrent IMRT of the chest at a prescription dose of 60-70 Gy. Primary endpoint was 1-year survival rate, and secondary endpoint was acute toxicity. Results The overall response rate was 68. 0％ and 72. 5％ in the PP and DP groups (χ2=0. 250, P=0. 617) . The median survival time was 19. 6 months ( 95％CI 13. 9-25. 3) versus 12. 1 months ( 95％CI 10. 7-13. 5) in the PP and DP groups. The 1-, 2-and 3-year overall survival rates were 72. 0％ versus 52. 9％, 28. 0％ versus 17. 6％, and 16. 0％ versus 13. 7％, respectively in the PP and DP groups ( P=0. 049) . In the PP and DP groups, the incidence of grade 3-4 leukopenia was declined by 48％ and 63％( P=0. 098) , and the incidence of grade 3-4 neutropenia was decreased by 34％ and 65％( P=0. 002) , the incidence of grade 3-4 anemia was reduced by 38％ and 10％(P=0. 024), and the incidence of grade 3-4 thrombocytopenia was declined by 40％ and 14％( P=0. 003) . The incidence rate of grade 2 pneumonitis ( P=0. 625) and grade 3 esophagitis ( P=0. 484) were similar in both groups. No patients experienced ≥grade 3 pneumonitis or ≥ grade 4 radiation esophagitis. Conclusions Pemetrexed-cisplatin combined with chemoradiotherapy yields higher clinical efficacy compared with docetaxel-cisplatin plus concurrent chemoradiation in the treatment of stageⅠV lung adenocarcinoma. The incidence of radiation pneumonitis and esophagitis is similar. The incidence and severity of hematological toxicity does not significantly differ between two groups.Treatment-related toxicity is tolerable in both groups. Clinical Trial Registration Chinese Clinical Trial Registry ( ChiCTR-TRC-13004184) .