Objective:A high performance liquid chromatography (HPLC) method was developed to determine the enzymatic activity of CD38 in blood, which was the major enzyme responsible for consuming nicotinamide adenine dinucleotide (NAD). Additionally, the study aimed to detect the differences in CD38 enzymatic activity among individuals of varying ages and health statuses.Methods:A 50 μl whole blood matrix and enzyme reaction substrate of 150 μl β-NAD at a concentration of 500 μmol/L were selected for the analysis. To eliminate the impact of endogenous β-NAD, the whole blood sample was pre-incubated at 37 ℃ for 20 minutes before adding the substrate. The reaction was terminated by perchloric acid (PCA) after incubation at 37 ℃ for 40 min. The change in product nicotinamide (NAM) before and after the enzymatic reaction was measured by HPLC to calculate the CD38 activity. The linearity, limit of detection, limit of quantification, precision, and stability of the method were evaluated. The CD38 enzymatic activities in 60 healthy volunteers and 30 colorectal cancer patients in blood were determined by the developed method.Results:Pre-incubation at 37 ℃ for 20 minutes eliminated the effect of endogenous β-NAD. The correlation coefficient of NAM was 0.999 in the concentration range of 0.1-3.2 μmol/L, with limit of detection of 0.5 nmol/L and limit of quantification of 2.1 nmol/L. The average within-run imprecision ( CV) and total CV were 3.22%-4.03% and 2.91%-4.70%, respectively. The recovery rate ranged from 94.82% to 96.81%. The CD38 activity of whole blood was stable by storage at 4 ℃ for 48 hours, storage at room temperature for 8 hours, thawing of frozen whole blood at room temperature for 2 hours, or repeated freeze-thawing three times. NAM, NAD standards, and pre-treatment samples were stable after 48 hours at 4 ℃ and 8 hours at room temperature. CD38 activity gradually decreased with increasing concentration of the added CD38 inhibitor 4-aminoquinoline derivative (78c). Measurement of 60 healthy physical examination population samples showed significantly higher CD38 enzyme activity in the elderly group than that in the young group ( t=-2.776, P=0.007) and measurement of 30 colorectal cancer patients showed significantly higher CD38 enzyme activity than that in healthy people ( t=-2.572, P=0.012). Conclusion:The established HPLC method for determining CD38 enzymatic activity is characterized by its simplicity, efficiency, accuracy, and reproducibility. This technique serves as a valuable tool for investigating aging and aging-related diseases.

Objective: To investigate the mediating effect of insomnia among mobile phone addiction, aggressive behaviors and self-perceived identity among university students. Methods: A total of 740 university students were sampled from five universities in Binjiang District, Hangzhou City using a cluster random sampling method. The mobile phone addiction, aggressive behaviors and self-perceived identity were assessed using the Mobile Phone Addiction Index Scale, the Athens Insomnia Scale, the Chinese college students' version of the Buss-Perry Aggression Questionnaire, and the Self-Perceived Identity Scale, and the mediating effect of insomnia among mobile phone addiction, aggressive behaviors and self-perceived identity was examined using Process macro program and Bootstrap method. Results: A total of 740 questionnaires were allocated, and 700 valid questionnaires were recovered, with an effective recovery rate of 94.59%. The respondents included 221 men (31.57%) and 479 women (68.43%), and there were 607 respondents with a specialty of medicine (86.71%). There were 331 participants detected with mobile phone addiction (47.29%), 90 with aggressive behaviors (12.86%) and 289 with low-level self-perceived identity (41.29%), and the prevalence rates of mild, moderate, severe and extremely severe insomnia were 28.00%, 26.14%, 26.43% and 19.43% among respondents, respectively. Mobile phone addiction had a direct predictive effect on aggressive behaviors (β=0.301, P<0.001) and self-perceived identity (β=-0.129, P<0.001), and presented an indirect predictive effect on aggressive behaviors (effect of mobile phone addiction on insomnia: β=0.083, P<0.001; effect of insomnia on aggressive behaviors: β=0.999, P<0.001; effect of insomnia on self-perceived identity: β=-0.698, P<0.001). The contributions of mediating effects caused by insomnia were 21.61% and 31.02% to total effects. Conclusions Insomnia presents partial mediating effects among mobile phone addiction, aggressive behaviors and self-perceived identity among university students. Mobile phone addiction may directly affect aggressive behaviors and self-perceived identity, and indirectly affect aggressive behaviors and self-perceived identity via insomnia.

Objective    To evaluate the effect of smoking and drinking status on the prognosis of patients with esophageal squamous cell carcinoma (ESCC). Methods    The clinical data of 483 patients with ESCC who underwent surgical treatment in Shannxi Provincial People's Hospital from 2007 to 2016 were retrospectively analyzed. Among them, 352 patients were male and 131 were female, with a median age of 64 (37-80) years. There were 311 smokers and 172 drinkers. The relationship between preoperative drinking or smoking status and the clinicopathological characteristics of patients with ESCC was analyzed. Log-rank method and Cox risk regression were used to conduct univariate and multivariate survival analysis, respectively. Results    The preoperative smoking status was related to the patient's tumor location (P=0.030). Drinking status was associated with tumor location (P=0.001), degree of differentiation (P=0.030), pathological T stage (P=0.024) and pathological N stage (P=0.029). Univariate survival analysis showed that smoking status did not affect the disease-free survival (DFS) (P=0.188) and overall survival (OS) (P=0.127) of patients with ESCC. However, patients who drank alcohol had worse PFS than non-drinking patients (29.37 months vs. 42.87 months, P=0.009). It was further proved that alcohol consumption was an independent risk factor affecting patients' recurrence and metastasis by using multivariate analysis (RR=1.28, P=0.040). Alcohol consumption also reduced the OS of patients by 21.47 months (P=0.014), however, multivariate analysis did not yield significant results. Conclusion    Preoperative drinking status is related to the stage and differentiation of patients with ESCC. It is an independent risk factor affecting the recurrence and metastasis of ESCC.

Objective To study the maturation and activation effects of hyaluronic acid (HA) modified polymer nanoparticles co-delivering adjuvants and antigens on mouse bone marrow dendritic cells (BMDCs). Methods HA-modified polylactic acid-glycolic acid copolymer (PLGA) and cationic lipid DOTAP were used as nanocarriers (DOTAP-PLGA) to co-deliver adjuvant CpG with model antigen ovalbumin (OVA). In the drug-loaded nanocarriers, CpG was covalently bound to the surface of HA, and OVA was physically blended into DOTAP-PLGA nanocarriers. The nanoparticles were characterized by transmission electron microscopy and dynamic light scattering. The in vitro release of CpG and OVA in the nanoparticles was investigated. The uptake and distribution of nanoparticles in mouse BMDCs were studied by flow cytometry and laser scanning confocal microscopy. The maturation and cytokine expression of mouse BMDCs were evaluated by flow cytometry and enzyme-linked immunosorbent assay, respectively. Results The CpG-HA-OVA-PLGA nanoparticles loading CpG and OVA were prepared. The average particle size was (305.1±2.2) nm and the polydispersity index was 0.203. A core-shell structure of the nanoparticles modified by HA was clearly observed by transmission electron microscopy. Cellular experiment results showed that CpG-HA-OVA-PLGA nanoparticles could be efficiently uptaken by mouse BMDCs, and promote lysosomal release of CpG and cytoplasmic delivery of antigen OVA. Compared with free OVA group and free OVA+CpG group, the CpG-HA-OVA-PLGA nanoparticles significantly up-regulated the expression of co-stimulatory molecules CD86 and CD40 (all P<0.01), major histocompatibility complex I (MHC-I) (P<0.01), and cytokine tumor necrosis factor-α (TNF-α) (P<0.01). Conclusions HA-modified CpG and OVA nanoparticle co-delivery vectors can effectively promote the maturation and activation of dendritic cells, which provides a basis for the development of novel vaccine vectors for the co-delivery of antigens and adjuvants.

Objective This study explored the expression of polyclonal protein SUZ12 in patients with intrahepatic cholangiocarcinoma(ICC),and its role in predicting the survival and treatment of ICC patients.Methods The expression of SUZ12 and p16INK4a was detected by immunohistochemical assay in 207 liver tissue samples including ICC patients,BilIN-1,-2,-3 and non-tumor-like cholangiocarcinoma.The expression of these proteins was assessed to be related to the pathological characteristics of the ICC patients receiving chemotherapy and the outcome of survival as well as the subsequent chemotherapy response.Results The expression level of SUZ12 was gradually increased from non-neoplastic bile duct tissue to BilIN-1,-2,-3 and ICC.The expression of p16INK4a protein was expressed in non-neoplastic-like cholangiocarcinoma,but it decreased gradually in BilIN-1,-2,-3 and ICC tissues.SUZ12 expression was associated with undifferentiated ICC,lymph node metastasis and advanced cancer.Kaplan-Meier curve analysis showed that ICC patients with high expression of SUZ12 had a significant reduction in overall survival and disease-free survival in comparison with ICC patients with the low expression of SUZ12.SUZ12 expression was significantly associated with overall survival of patients receiving adjuvant gemcitabine-based chemotherapy(AGC).Conclusion SUZ12 expression is able to predict the overall survival and disease-free survival of ICC patients with adjuvant gemcitabine-based chemotherapy.

Objective    To investigate the efficacy of Docetaxel injection and Capecitabine tablets combined with Oxaliplatin injection in chemotherapy for patients after esophageal cancer surgery. Methods    We retrospectively analyzed the clinical data of 101 patients with esophageal cancer who underwent radical surgery from June 2010 to December 2012, including 58 males and 43 females. According to the different treatment methods they were divided into a study group (58 patients, 32 males and 26 females, postoperatively receiving Docetaxel injection, Capecitabine tablets, Oxaliplatin injection and chemotherapy) and a control group (43 patients, 26 males and 17 females, taking Docetaxel injection and Capecitabine tablets for 4 consecutive courses). We compared the difference in the outcomes between the two groups. Results    There was no significant difference in the level of serum anticancer antigen (CEA), carbohydrate antigen 125 (CA125), carbohydrate antigen 199 (CA199) and squamous cell carcinoma antigen (SCC) before chemotherapy between two groups (P>0.05). After chemotherapy, the level of serum CEA, CA125, CA199, SCC in the study group was significantly lower than that in the control group (P<0.05). The 1-year survival rate of the study group was 92.59% and the 2-year survival rate was 70.37%, which were not significantly different from those of the control group  (P>0.05). The 3-year survival rate of the study group was significantly higher than that of the control group (57.41 % vs. 32.43%, P<0.05). The mean survival time of the study group was longer than that of the control group (31 months vs. 22 months, P=0.001). Conclusion    Docetaxel injection and Capecitabine tablets combined with Oxaliplatin injection for the treatment of esophageal cancer surgery can significantly reduce levels of tumor markers in serum after esophageal cancer surgery, and is favorable for the long-term survival of patients, but adverse reactions should be noted.

Objective To explore the changes of the constituent ratio of hypoglycemic scheme and clinical outcomes of patients with type 2 diabetes mellitus(T2DM)in recent three years in Shihezi. Methods The cluster random sampling methods were used to select 300 patients with T2DM who met the standards in November 2012 from 13 communities in Shihezi. The datasets including general demographic information ,treatment and clinical outcomes were collected by following them up for three years. Results From 2012 to 2015,the proportion of pa-tients with oral drugs decreased from 63.5％ to 51％ while increased from 30.8％ to 41.8％ with insulin treatment. For the patients with insulin treatment ,the rate of patients on single drug therapy declined significantly (χ2 =8.77,P<0.05),while significantly increased on insulin combined with oral drug(χ2=-10.27,P<0.01). The incidence of adverse effects increases from 16.8％ to 24.5％. As compared with 2012,blood sugar levels and con-trol rate had no obvious changes in 2015;namely,according to the standard(1),the control rate of blood glucose in 2015 was 41.2％,decreasing 4.0％as compared with 2012,while according to the standard(2),it increasd by 1.4％ from 2012 to 2015(52.9％). The rate of diabetic complications significantly increased from 2012 to 2015. Conclusions Oral drugs are mainly used in the treatment of T2DM in Shihezi communities,whereas the rate of insulin use elevates. The blood glucose control rate,medication safety,and ability to lower the rate of diabetic com-plications need to be improved in T2DM patients in Shihezi communities.

In this article, the authors expounded the necessary and problems of animal experiment post-ap-proval monitoring ( PAM) , which included who should do PAM, how to do PAM, and what the PAM can do. The authors also exposed the following suggestion: formulating the detailed rules, regulations and SOP, strengthening the training of PAM team member and animal experiment personnel, and monitoring the whole process of animal protocol review using the software.

Objective: To obtain purified and functional CDNF-his recombinant protein and prepare its polyclonal antibodies.Methods:Preparation of recombinant CDNF-his was carried out in HEK 293 T cells with pVR1012-CDNF-his successfully constructed transfected into them.Then,the recombinant protein was purified by Ni-NTA immunoaffinity chromatography.The purity was analyzed by SDS-PAGE and the protein′s identity was tested by Western blot.MTT was used to verify the biological function of the protein purified.New Zealand white rabbits were immunized with purified CDNF-his protein for preparation of polyclonal antibodies.Results:pVR1012-CDNF-his expressed successfully in HEK 293 T cells.The purity of protein was up to more than 90%after purification.MTT showed that CDNF-his was able to protect PC 12 cells from damage by 6-OHDA.The polyclonal antibody was detected at the end of animal immunizing process.Conclusion: A method to express and purify protein using HEK 293T cell and following Ni-NTA immunoaffinity chromatography has been built.CDNF-his with biological activity is obtained based that.Finally, polyclonal antibodies of CDNF were generated successfully.

Objective To explore the relationship of X-ray exposure parameters and false-node rate during image-guidance treatment with CyberKnife spine tracking.Methods Using spine tracking planning on a chest phantom,several combinations of X-ray exposure parameters were used to locate.The false-node ratio and the surface absorbed dose were investigated and the radiation dose was optimized.Results The false-node ratio and surface absorbed radiation dose decreased when the X-ray exposure parameters increased until they saturated.In the range of ≤5.0％ false-node rate,the surface absorbed radiation dose was 0.11,0.26 mGy,and 0.31-0.46 mGy,when the false-node rate was 2.77％,1.07％and 1.0％,respectively.Conclusions In image-guided treatment of CyberKnife,the radiation dose would be optimized,and the patient's radiation dose would be reduced greatly,which is important to protect the patients.