Objective To purify the anti-T cell immunoglobulin mucin ( TIM)-3 monoclonal antibody 4E8 and examine its biological function in vitro. Methods The mouse monoclonal antibody against human TIM-3, clone 4E8, was obtained by standard protocol for monoclonal antibody purification. The cell lines expressing human TIM-3 molecule were obtained by cell transfection technique. We ex-amined the ability of 4E8 binding to human TIM-3 by flow cytometry. The ability of 4E8 blocking the binding of fusion protein TIM-3 Ig-huFc with phosphatidylserine( PtdSer) , the apoptotic cell surface TIM-3 ligand, was also analyzed by flow cytometry. Mixed lympho-cyte reaction ( MLR) and ELISA assays were used to determine the effect of TIM-3 monoclonal antibody ( 4E8) on IFN-γsecretion in CD4+ T cells. Results 4E8 specifically bound to human TIM-3 but could not block the binding of TIM-3 to Ptdser. Compared with the negative control (IFN-γ secretion: 958.3±153.2), 4E8 enhanced the ability of CD4+ T cells to secrete IFN-γ in MLR (4E8 of 10μg/mL group:IFN-γ secretion 2563±150.3 and 4E8 of 3.33 μg/mL group:IFN-γ secretion 1981±211.5) with statistically signifi-cant difference ( P<0.05) . In addition, the combined application of 4E8 with the anti-programmed death-1 ( PD-1) monoclonal anti-body nivolumab showed synergistic effects for increasing IFN-γ secretion in MLR assay ( 4E8 of 10 μg/mL group: IFN-γ secretion 3049±80.5 and 4E8 of 0.33μg/mL group:IFN-γsecretion 1957±321.3) as compared with 4E8 alone (10μg /mL group:IFN-γse-cretion 2563±150.3 and 0.33 μg/mL group:IFN-γ secretion 844±76.2) with statistically significant difference (P<0.05). Conclu-sion We successfully obtained a 4E8 clone of monoclonal antibody to human TIM-3 which may enhance the capacity of IFN-γsecre-tion from CD4+ T cells. The effect of enhancing IFN-γ secretion of CD4+T cells by TIM-3 monoclonal antibody was independent from blocking the binding of TIM-3 with Ptdser.

Objective To investigate the feasibility of the highGresolution MRI (HRGMRI)in diagnosing the intracranial artery dissection (IAD)and to improve radiological diagnostic accuracy in clinical practice.Methods The clinical data and imaging data of 43 patients with suspected IAD were analyzed retrospectively.HRGMRI and DSA examinations were performed.HRGMRI images were reviewed by two experienced radiologists independently,and DSA images were analyzed by a neurosurgeon.FollowGup durations were from 3 months up to 12 months,and then the clinical diagnoses of IAD were made.The sensitivity,specificity,positive predictive value (PPV),and negative predictive value (NPV)of HRGMRI were calculated,and the consistency of HRGMRI and DSA in the diagnosis of IAD was made.Results 32 over 43 patients were detected to be positive by HRGMRI,however,34 of 43 patients were diagnosed as IAD clinically.The sensitivity and specificity of HRGMRI for detecting IAD were 9 1.2% and 88.9%,respectively.The PPV was 96.9%,and the NPV was 72.7%.For DSA,30 patients with IAD were detected and 1 3 patients had negative results.Compared with the HRGMRI (3 2/43),the coincidence rate of two examinations was 86.0%.McNemer test (P＝0.687)indicated that there was no significant difference between two methods.Conclusion HRGMRI has excellent soft tissue contrast and spatial resolution.As a nonGinvasive modality with good tissue contrast and spatial resolution,it plays an important role in the diagnosis,differential diagnosis and followGup of IAD.

Objective:To investigate the effect of Lee Silverman voice therapy (LSVT) on the speech and life quality of Chinese persons with Parkinson′s disease (PD).Methods:A total of 16 Chinese PD patients were enrolled and given standard LSVT for 4 weeks. Their acoustic data were analyzed using PRAAT software before and after the treatment. Their voice quality was evaluated using the Japanese GRBAS voice scale and their quality of life was quantified using the Voice Handicap Index (VHI) scale.Results:After 4 weeks of LSVT the patients′ average voice and life quality had improved significantly. The average maximum duration of sustained vowel phonation had increased significantly, as had the vowels′ mean loudness, reading and monologue delivery. The GRBAS grading indicated that hoarseness had decreased significantly. The average total voice handicap index had also decreased significantly.Conclusion:Lee Silverman therapy can significantly improve maximum phonation time and loudness of Chinese-speakers, which enhances sensory feedback. The quality of their speech and of their life also improve significantly. This technique is recommended for clinical application.

The fusion neutron source based on Gas Dynamic Trap (GDT) device has the advantages of high neutron flux, large testing space and low construction cost, so it is an ideal solution for high-flux fusion volumetric neutron source. As a new type of radioactive facility, how to carry out radiation safety management under the current nuclear safety regulatory laws and regulations in China is an important issue that must be addressed before the construction of this facility. In this paper, the radiation safety characteristics of GDT fusion neutron source was analyzed and the main radiation terms of GDT fusion neutron source were expounded. Combined with the existing nuclear safety regulatory laws and regulations in China, the key points of radiation protection and safety management in the process of construction and application of GDT fusion neutron source were put forward. It is recommended that the GDT neutron source should be managed as a class I radiological device. Prior to operation, radiation safety permit and nuclear material permit should be obtained, and special attention should be paid to the safety protection of radioactive tritium. This study will pay the way for the radiation safety management of the international high flux fusion neutron source ALIANCE project, and also provide the reference for the related nuclear and radiation safety management departments in China.

Objective To establish a method of LC-MS/MS for determining cordycepin(Cor)and 3′-deoxyinosine(3′-Deo)concentration in rat plasma,and to study their pharmacokinetics in rats.Methods Protein was precipitated with methanol using 2-chloadenosine(2-Chl)as an internal standard.The chromatography was performed on Kinetex C18(3 mm×100 mm,2.6 μm,Phenomenex,USA)with gradient elution in aqueous(5 mmol·L-1 ammonium acetate)-methanol solution as mobile phase.ESI ion source was used for mass spectrometry,and positive ion multiple reaction monitoring(MRM)was used for scanning detection.The pharmacokinetics of Cor and 3′-Deo after oral administration of Cor(10 mg·kg-1)were studied in rats.Results Cor at 0.5-100 ng·mL-1 and 3′-Deo at 1-200 ng·mL-1 had good linearity,and the lower limits of quantification were 0.5 and 1 ng·mL-1,respectively.After oral administration of Cor in rats,the plasma concentration of Cor was low,which was mainly converted into the metabolite 3′-Deo.The Cmax of Cor and 3′-Deo were(5.4±3.4)and(142.0±50.0)ng·mL-1,and AUC0-360min min were(658.4±459.3)and(18 034.9±4 981.1)ng·min·mL-1,respectively.Conclusion The method is simple,sensi-tive,and accurate,which is suitable for determining Cor and 3′-Deo concentration in plasma and the pharmacokinetic study.

Objective:To analyze the role of patient support system in multidrug-resistant/rifampicin-resistant tuberculosis (MDR/RR-TB) treatment adherence and outcome, and provide evidence for deepening the patient-centered support system.Methods:Based on the stakeholder analysis, definite stakeholders (administrators from the Department of Medical Insurance, and those from the provincial CDC), expectant stakeholders (administrators from regional CDC, health workers from primary CDCs, medical workers from designated MDR/RR-TB hospitals and MDR/RR-TB patients), and latent stakeholders (MDR/RR-TB patient families and their neighbors or colleagues) were selected using a purposive sampling. These stakeholders were subject to a semi-structured interview on patient support. The inclusion of participants ceased after reaching code or thematic saturation and meaning saturation, while thematic framework analysis was applied in interview data.Results:The 25 interviewees included could be categorized into three groups of stakeholders, i. e., 4 definite stakeholders, 19 expectant stakeholders and 2 latent stakeholders. Three themes summarized in this regard were definite stakeholders providing policy support to advance these patients′ access to standardized diagnosis and treatment services; diagnosis and treatment and management support of expectant stakeholders of these patients to encourage their compliance to treatment and enable their access to high quality medical care; and support from latent stakeholders as a critical guarantee for the patients to welcome a desirable treatment outcome. Psychological support provided under MDR/RR-TB basic care program in some provinces contributed positively to raising patients′ compliance. Economic support, treatment support from family menmbers ccould help the patients to welcome desirable outcomes.Conclusions:MDR/RR-TB patient-centered support system operating in the Yangtze River delta provide the patients with MDR/RR-TB diagnostic and treatment services of some accessibility. Given the progress, there are still shortcomings for the respective stakeholders to enhance their attention and collaboration to improve the access and equity to medical service.

Objective: To investigate the expression of IFIT2 (interferon-induced protein with tetratricopeptide repeats 2) in human lung cancer tissue and analyze the relationship between the IFIT2 expression and clinicopathological features and prognosis. Methods: Tissue microarray and immunofluorescence staining were used to examine the IFIT2 expression in lung cancer tissue and their adjacent tissues. Wilcoxon rank test was used to compare the IFIT2 expression in lung cancer and corresponding adjacent tissues. The chi-square test was used to analyze the relationship between the IFIT2 expression in lung cancer tissues and clinicopathological features of the patients. Kaplan-Meier survival analysis was performed to analyze the correlation between IFIT2 expression and patients′ overall survival. Cox model was used to analyze the correlation between different clinical parameters and prognosis. Results: There was significant difference for the IFIT2 expression between the lung cancer tissues and adjacent tissues (P＜0.01). There was no significant correlation between IFIT2 expression and clinicopathological features of patients (P＞0.05). In lung adenocarcinoma and squamous cell carcinoma, Kaplan-Merier survival analysis showed that the overall survival (OS) of patients in IFIT2 low expression group was significantly shorter than that in IFIT2 high expression group (HR=2.392, 95%CI: 1.103-5.186, P=0.027; HR=2.907, 95%CI: 1.118-7.559, P=0.029, respectively). Multi-factor Cox model analysis indicated that distant metastasis (HR=8.033, 95% CI: 3.664-17.614, P=0.000) was independent prognostic factors for lung adenocarcinoma, lymph node metastasis (HR=3.390, 95% CI: 1.029-11.175, P=0.045) and IFIT2 low expression (HR=3.762,95%CI: 1.236-11.451, P=0.020) were independent prognostic factors for lung squamous cell carcinoma. Conclusion The down-regulated expression of IFIT2 in lung cancer tissues suggests that it may play an important role in initiation and development of lung cancer. It could be used as a valuable risk factor to predict the prognosis of lung cancer patients.

Objective To investigate the neural mechanisms of speech motor control in individuals with Parkinson's disease (PD) under different strategies of motor execution control.MethodsThe techniques of the psychoacoustic and event-related potentials (ERPs) based on the altered auditory feedback paradigm were used in the present study. Two groups, including 17 PD patients and 17 healthy controls, were instructed to produce sustained vowels under the internal and external cueing tasks while hearing their voice randomly pitch-shifted downwards. The vocal responses and associated ERPs were recorded and compared across the groups and tasks.ResultsBehavioral results showed that the amplitude of acoustic compensation response was larger in PD patients than in the healthy controls (F=5.415, P=0.027), however, the main effect was not significant in the tasks (F=0.039, P=0.840). At the cortical level, PD patients produced significantly larger N1 responses to pitch perturbations in the internal cueing task related to the external cueing task (F=8.634, P=0.006), while such task effect was not observed in the healthy controls (F=1.550, P=0.231). Also, PD patients produced significantly larger N1 responses than the healthy controls in the internal cueing condition (F=5.476, P=0.026), but not in the external cueing condition (F=0.249, P=0.621). Conclusion Speech motor control in PD can be influenced by the strategies of motor execution control. Compared to the internal cueing task, the external cueing task can increase neural efficiency in the encoding of speech auditory feedback PD patients that improve auditory-motor integration for speech production.

Objective: To investigate the degree of infiltration and distribution of tissue-resident CD8 + T cells (CD103 + CD8 + T cells) in gastric cancer tissues, and analyze the relationship between the degree of infiltration and clinicopathological features and prognosis. Methods: Tissue microarray and immunofluorescence staining were used to examine the CD8 + T cells and CD103 + CD8 + T cells infiltration in 90 cases of gastric cancer and their adjacent normal tissues. Wilcoxon rank test was used to compare the CD8 + T cells, CD103 + CD8 + T cells infiltration and CD103 + CD8 + T cells/CD8 + T cells ratio in gastric cancer and corresponding normal tissues. The chi-square test was used to analyze the relationship between CD8 + T cells, CD103 + CD8 + T cells infiltration and CD103 + CD8 + T cells/ CD8 + T cells ratio in gastric cancer tissues and clinicopathological features of the patients. Kaplan-Meier survival analysis was performed to explore the correlation between CD8 + T cells, CD103 + CD8 + T cells infiltration and CD103 + CD8 + T cells/ CD8 + T cells ratio and overall survival. Cox model was used to analyze the correlation between different clinical parameters and prognosis of the patients. Results: There was no significant difference for the infiltration of CD103 + CD8 + T cells between the gastric cancer tissues and adjacent normal tissues (P＞0.05). The infiltration rate of CD103 + CD8 + T cells in the cases in stage Ⅲ to Ⅳ (69.09%, 38/55) was significantly lower than that in the stage Ⅰ to Ⅱ cases (91.43%, 32/35), (χ 2 =6.175, P=0.013). There was no significant correlation between CD103 + CD8 + T cells infiltration and other clinicopathological features (P＞0.05). Kaplan-Meier survival analysis showed that the patients with high CD103 + CD8 + T cells infiltration showed significantly longer overall survival than the patients with low CD103 + CD8 + T cells infiltration (HR=2.187, 95%CI: 1.062-4.500, P=0.033 6). Multivariate Cox model analysis indicated that tumor diameter (HR=2.031, 95%CI: 1.163-3.546, P=0.013) and CD103 + CD8 + T cells infiltration (HR=0.516, 95%CI: 0.285-0.934, P=0.029) were independent prognostic factors for gastric cancer. Conclusion CD103 + CD8 + T cells in gastric cancer tissues should be associated with good prognosis, suggesting that they play an important role in the inhibition of gastric carcinogenesis and development, and can be used as an important factor for the prognosis evaluation of the patients with gastric cancer.

Objective: To investigate the degree and distribution of tissue-resident CD8+ T cell (CD103+CD8+T cells) infiltration in colorectal cancer (CRC) tissues, and to analyze its relationship to patients’clinicopathological features and prognosis. Methods: Tissue chips of 88 cases of colon cancer tissues (No.HColA180Su14) and 77 cases of rectal cancer tissues (No. HRec-Ade180Sur-03) were obtained from Shanghai Outdo Biotech Co.,Ltd. Immunofluorescence staining was performed to examine the infiltration pattern and degree of CD103+CD8+T cells in the collected CRC tissues and their para-cancerous tissues. Wilcoxon rank test was used to compare CD103+CD8+T cell infiltration degree in CRC tissues and the para-cancerous tissues. Chi-square test was used to analyze the relationship between CD103+CD8+T cell infiltration in CRC and patients’clinicopathological features. Kaplan-Meier survival analysis was conducted to explore the correlation between CD103+CD8+T cell infiltration and patients’prognosis. Cox model was applied to analyze the correlation between different clinical parameters and patients’prognosis. Results: CD103+CD8+T cell infiltration presented no signifi ·cant differences between CRC and para-cancer tissues (P>0.05). Patients with distant metastasis had significantly lower CD103+CD8+T cell infiltration rate than patients without distant metastasis (P<0.01). There was no significant correlation between the infiltration of CD103+CD8+T cells and other clinicopathological features (P>0.05). Kaplan-Meier survival analysis showed that the overall survival (OS) of patients with high CD103+CD8+T cell infiltration was significantly longer than that of the patients with low infiltration (54.42% vs 25.00%, P<0.05). Multivariate Cox model analysis indicated that pathological grade (P<0.01) and high CD103+CD8+T cell infiltration (P<0.05) were independent prognostic factors for CRC. Conclusion： ：CD103+CD8+T cell infiltration in CRC is associated with patients’prognosis, suggesting that CD103+CD8+T cell plays an important role in the initiation and development of CRC.