Objective:To translate the Cardiac Symptom Survey and analyze its reliability and validity in the post-valvular heart replacement survey and to provide the basis for the clinical application of this scale.Methods:The symptoms of 233 patients after heart valve replacement were investigated in Suining Central Hospital by Chinese version of the Cardiac Symptom Survey. The reliability of the scale was evaluated by Cronbach'α coefficient and the split half coefficient, the validity of the scale was evaluated by exploratory factor analysis and content validity index.Results:The internal consistency coefficient of the Chinese version of the Cardiac Symptom Survey ranged from 0.875 to 0.968, both above 0.870.The split half coefficient was 0.699,which demonstrated better internal consistency.The content validity index of the Chinese version of the Cardiac Symptom Survey ranged from 0.84 to 1.00.Factor analysis got five factors, which explained 68.133% of the total variance.Conclusions:The Chinese version of the Cardiac Symptom Survey has good reliability and validity in the investigation of the symptoms of patients after heart valve replacement, and can be used for the investigation and management of the symptoms of patients after heart valve replacement.

Objective:To investigate the regulation and mechanism of chronic Trichinella spiralis ( Ts) infection on liver immunopathology in mice co-infected with Plasmodium berghei ANKA( PbA). Methods:According to body weight, 64 specific pathogen free female Kunming mice (6 - 8 weeks old, weighting 22 - 25 g) were divided into 4 groups by using random number table method. Control group: uninfected; Ts group: mice were mono-infected with 30 Ts larvae by oral gavage on day 0; PbA group: mice were mono-infected with 1 × 10 6PbA-infected red blood cells in 0.1 ml of phosphate buffer (PBS) administered by intraperitoneal injection on day 121; co-infected ( Ts+PbA) group: mice were infected with 30 Ts larvae by oral gavage and intraperitoneal injected with 1 × 10 6PbA-infected red blood cells in 0.1 ml PBS on day 121 after Ts infection. There were 16 mice in each group, in which 10 mice in each group were monitored for the survival rate. The peripheral red blood cell parasitemia of PbA group and Ts + PbA group were monitored every other day by light microscope examination of Giemsa-stained thin tail-blood smears from day 3 after PbA infection. Mice were sacrificed at day 135 after Ts infection and/or at day 15 after PbA infection, the mouse body weight and liver weight were measured, and the liver index were calculated. Ts-infected mice were monitored by a light microscope examination of diaphragm compression slide. Under a light microscope, the liver pathology and liver fibrosis of mice were observed and compared with hematoxylin-eosin (HE) staining and Sirius red staining, respectively. The F4/80 + Kupffer cells in liver of mice were examined by immunohistochemical staining. Results:After infection with Ts or PbA, Ts larvae cysts were observed in diaphragm tissues and PbA were observed in red blood cells under the light microscope. After PbA infection, there was no significant difference in survival rate between PbA group and Ts+ PbA group ( P > 0.05). Compared with PbA group, the peripheral red blood cell parasitemia was significantly decreased in Ts+ PbA group on days 11 and 15 after PbA infection (%: 27.104 ± 7.623 vs 45.032 ± 9.849, 60.218 ± 2.776 vs 76.778 ± 6.351, P < 0.05), and the liver index and the liver pathology score were significantly decreased in Ts+ PbA group ( P < 0.05). Sirius red staining showed that the positive area of liver fibrosis in Ts+ PbA group was significantly higher than that in PbA group ( P < 0.05). Immunohistochemical staining showed that the average optical density value of F4/80 + Kupffer cells in Ts+ PbA group was significantly higher than that in PbA group ( P < 0.01). Conclusion:Chronic Ts infection may reduce the peripheral red blood cell parasitemia, increase F4/80 + Kupffer cells expression in liver, and attenuate liver pathology in mice co-infected with PbA.

The fragment of MDR1 gene obtained from the plasmid pHaMDR1-1 carrying the whole human MDR1 cDNA, was cloned reversely into the shuttle plasmid pAdTrack-CMV. With the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the Escherichia coli BJ5183 and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged in the 293 cells. The recombinant adenovirus MDR1 vector would introduce the antisense MDR1 gene into the human multidrug resistance hepatocellular cell line effectively, which would provide an experimental basis for studies on the multidrug resistance in human hepatocellular carcinoma.
ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; Adenoviridae ; genetics ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; genetics ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; genetics ; Escherichia coli ; genetics ; Gene Transfer Techniques ; Genes, MDR ; genetics ; Genetic Vectors ; Humans ; Liver Neoplasms ; drug therapy ; genetics ; Multidrug Resistance-Associated Proteins ; biosynthesis ; genetics ; Oligonucleotides, Antisense ; pharmacology ; Plasmids ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic

Objective To evaluate the efficacy and safety of recombinant adenovirus-p53(rAdp53) injection combined with radiotherapy and hyperthermia in the treatment of unresectable advanced soft tissue sarcoma.Methods In this retrospective study, we evaluated 76 patients with unresectable advanced primary or recurrent soft tissue sarcoma treated in our hospital from November 2005 to November 2012.These patients received radiotherapy and hyperthermia with rAdp53(p53 group, n=41) or without rAdp53(control group, n=35).rAdp53((1-2)×1012viral particles each time, once a week, 8 times on average) was injected into the tumor or infused into the pelvic cavity.Radiotherapy (2 Gy each time, 5 times a week) was performed for the planning target volume at 56.3±5.3 Gy in the p53 group and 58.1±4.2 Gy in the control group, with no significant difference between the two groups (P>0.05).Superficial or deep thermotherapy was employed 8 times on average (twice a week).Clinical features, response rate, time to progression (TTP), overall survival (OS), and adverse events were compared between the two groups (P>0.05).The Kaplan-Meier method was used to calculate OS;the log-rank test was used for survival difference analysis and univariate prognostic analysis;the chi-square test was used for comparison of categorical data.Results At 2 months after treatment, the p53 group had significantly increased response rate (partial response+ complete response+ stable disease)(85% vs.54%, P=0.003) and local control rate (49% vs.23%, P=0.020) as well as prolonged TTP (12 months vs.5 months, P=0.010) and OS (48 months vs.31 months, P=0.049), as compared with the control group.No adverse events caused by radiotherapy and hyperthermia except transient fever were seen in the two groups.Conclusions Concurrent radiotherapy and hyperthermia combined with rAdp53 injection is effective and safe for patients with advanced soft tissue sarcoma.

Objective:To develope a self-adjustable automatic planning method of intensity modulated radiotherapy based on predicted dose, in order to enhance the robustness of automatic planning.Methods:After the patients′ dose by 3D U-Res-Net_B network was predicted, the current dose was calculated based on the last iteration result, then the predicted dose was combined to calculate the target dose and optimized. With all iterations completed or exit conditions satisfied, final treatment plannings would be acquired. A total of 30 cases of rectal cancer were tested to verify the effectiveness of the algorithm.Results:The mean value of planning target volumes′ V100% was (95.03±0.91)% for clinical plans, close to (94.67±1.96)% for automatical plans( P>0.05), and better than (92.90±2.13)% for predicted dose with the statisically significant difference ( t=29.0, P<0.05). Automatic planning′s indexes such as V35 of small intestines, V40 of bladders and V20 - V40 of femoral heads were lower than predicted and clinical ones, with the statisically significant difference( t=4.5-118.0, P<0.05). Discrepancy in other indexes of organs at risk was not statistically significantly different( P>0.05). Conclusions:This method made automatic planning processes more robust and more adaptive to difficult clinical situations.

Objective:To develop a dose prediction-based quantitative evaluation method of the quality of radiotherapy plans, and to verify the clinical feasibility and clinical value of the method .Methods:The 3D U-Netwas trained using the radiotherapy plans of 45 rectal cancer cases that were formulated by physicists with more than five years of radiotherapy experience. After obtaining 3D dose distribution using 3D U-Net prediction, this study established the plan quality metrics of intensity modulated radiotherapy(IMRT) rectal cancer radiotherapy plans using dose-volume histogram(DVH) indexes of dose prediction. Then, the initial scores of rectal cancer radiotherapy plans were determined.Taking the predicted dose as the optimization goal, the radiotherapy plans were optimized and scored again. The clinical significance of this scoring method was verified by comparing the scores and dosimetric parameters of the 15 rectal cancer cases before and after optimization.Results:The radiotherapy plans before and after optimization all met the clinical dose requirements. The total scores were(77.21±9.74) before optimization, and (88.78±4.92) after optimization. Therefore, the optimized radiotherapy planswon increased scores with a statistically significant difference( t=-4.105, P<0.05). Compared to the plans before optimization, the optimized plans show decreased Dmax of all organs at risk to different extents. Moreover, the Dmax, V107%, and HI of PTV and the Dmax of the bladder decreased in the optimized plans, with statistically significant differences ( t=2.346-5.771, P<0.05). There was no statistically significant difference in other indexes before and after optimization ( P>0.05).The quality of the optimized plans were improved to a certain extent. Conclusions:This study proposed a dose prediction-based quantitative evaluation method of the quality of radiotherapy plans. It can be used for the effective personalized elevation of the quality of radiotherapy plans, which is beneficial to effectively compare and review the quality of clinical plans determined by different physicists and provide personalized dose indicators. Moreover, it can provide great guidance for the formulation of clinical therapy plans.

OBJECTIVE: To express and purify NR4A1-DNA binding domain (DBD) protein of nuclear receptors. METHODS: The fusion protein PET28a-NR4A1-DBD was constructed and purified with the nickel affinity chromatography, cation-exchange chromatography and gel filtration chromatography. RESULTS: The protein PET28a-NR4A1-DBD was mostly soluable at 24 °C. A total of 2-3 mg/L pure NR4A1 proteins were yielded in bacterial culture and the purity for final fractions of NR4A1-DBD protein were great than 95% by SDS-PAGE analysis. CONCLUSION Nickel affinity chromatography is effective to purify protein. The protein purity can be further improved by the following methods including cation-exchange chromatography and gel filtration chromatography.
Electrophoresis, Polyacrylamide Gel ; Nuclear Receptor Subfamily 4, Group A, Member 1 ; chemistry ; Recombinant Fusion Proteins ; chemistry

OBJECTIVETo optimize the protocols for isolation and culture of mesenchymal stem cells from rat bone marrow (BMSCs).

METHODSBMSCs were isolated by adherence to plastic with frequent medium change and reduced trypsinization time. The cell growth curves were drawn and the surface markers of BMSCs were detected by flow cytometry. The cells were induced to differentiate into osteogenic, adipogenic, hepatic and cholic lineages.

RESULTSThe cells isolated using this method were positive for CD29, CD44, and CD90 and negative for the hematopoietic surface markers CD45. The osteogenic and adipogenic differentiation of the BMSCs was verified by alkaline phosphatase staining, Alizarin red staining and Oil red staining. The cell subcultures up to passage 10 maintained capacities of differentiation into osteogenic and adipogenic lineages. The BMSCs induced with sequential addition of growth factors, cytokines and hormones differentiated into cells expressing hepatocyte- and cholangiocyte-specific markers.

CONCLUSIONThe optimized method allows efficient isolation of homogenous populations of MSCs from rat bone marrow, which can be induced into multiple cell lineages.

Animals ; Cell Culture Techniques ; Cell Differentiation ; Cell Proliferation ; Cell Separation ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Rats

Objective To review the failure patterns and clinical outcomes for patients with cervical esophageal carcinoma (CEC) undergoing definitive radiotherapy (RT).Methods Medical records,clinical characteristics and outcomes of patients with CEC treated by definitive RT from August 2008 to May 2017 were retrospectively reviewed and analyzed.Results A total of 97 patients with squamous cell CEC were enrolled in this study with a median age of 59 years old (range 18-78 years old).There were 34 patients with limited cervical esophagus,and 63 patients with diseases beyond cervical region,respectively.There were 69,7,and 6 patients with Bronchi invasion,thyroid lobes involvement and aortic involvement,respectively.There were 11,80 and 6 patients with stage Ⅱ,Ⅲ and Ⅳ (non-regional lymph node metastases),respectively.The median dose to the gross tumor volume (GTV) was 66 Gy,in which 46 patients received above 66 Gy and 51 patients received less than 66 Gy,respectively.The median progression free survival (PFS) and overall survival (OS) were 16.03 and 23.30 months,respectively,with a median follow-up of 14.90 months.The 1,2,3-year PFS and OS were 56.86％,30.35％,26.34％,and 72.54％,47.94％,40.81％,respectively.Sixty-one patients had treatment failure at their last follow-up,in which 40,27,and 18 patients developed local failure,regional failure,and distant metastasis,respectively.Univariate analysis revealed that thyroid lobes involvement resulted in lower PFS (x2 =5.773,P＜0.05) and OS (x2 =13.461,P＜0.05),and bronchi involvement (x2 =4.283,P＜0.05) was associated with lower OS.Multivariate analysis indicated that aortic involvement and thyroid lobes involvement were associated with lower PFS (x2 =6.796,4.548,P＜0.05) and OS (x2 =13.421,10.581,P＜0.05),and GTV dose above 66 Gy was associated with higher OS (x2=5.296,P＜0.05).Conclusions Local-regional recurrence was the main failure pattern for patients with CEC after definitive RT.Aortic,thyroid lobes,and/or bronchi involvement were associated with poor prognosis,and GTV dose ≥66 Gy tended to improve OS.Prospective studies with larger population were needed to further confirm this study.