Objective To study the antigenicity of RNA polymerase and the relationship between the protein and the SARS-CoV replication. Methods The fragment was amplified by PCR, ligated with the prokaryotic expression vector pET32a+ and transformed into E. coli BL-21 .The expressed fusion protein identified by Western-blot and ELISA was used to detect the anti-SARS CoV IgG in different sera. Results The fusion protein was expressed successfully in E. coli BL-21. Detected by ELISA, the positive percentage of the anti-SARS IgG in the healthy donors, patients, infected animals and the rhesus administrated with the inactivated-virus were 0%, 95%, 100% and 0%, respectively. Conclusion Good antigenicity was shown in the expressed RNA polymerase. It can be used to diagnose the infection and to demonstrate the replication of SARS-CoV virus.

Objective To estimate the dose-response relationship between sedentary behavior with mortality in patients with type 2 diabetes. Methods A total of 17786 type 2 diabetic patients were recruited as participants, who were included in National Basic Public Health Service in Changshu County of Suzhou City, Qinghe District and Huai'an District in Huai'an City of Jiangsu Province. Cox proportional hazards regression model and restricted cubic spline model were employed to estimate the dose-response relationship between sedentary behavior with all-cause and cause specific mortality in patients with type 2 diabetes. Results Among 78114.34 person-years of the fo1low-up, the median of follow-up time was 4 years, and 1285 deaths occurred during that period. Compared to patients with sedentary behavior≤2 h/d, the multivariate adjusted hazard ratios of all-cause death associated with sedentary behavior levels of 3-4 h/d, 5-6 h/d, and≥7 h/d were 1.05(95％CI 0.92-1.20), 1.20(95％CI 1.03-1.42), and 1.39 (95％CI 1.16-1.65), respectively. Eevry increase of 1 h/d in sedentary behavior was associated with an increased hazard of death from cardiovascular disease(CVD) of 4％(HR=1.04, 95％CI 1.01-1.07) and from other causes of 6％( HR=1.06, 95％CI 1.03-1.09) . However, no significant association between sedentary behavior and malignant tumor death was found. The multivariable restrictive cubic spline regression indicated that the linear dose-response relationships were found between sedentary time with the all-cause, CVD cause, and other cause of mortality ( Non-linear test, P>0.05) . Conclusion Longer sedentary behavior could increase the risk of mortality in patients with type 2 diabetes.

Objective:To investigate the incidence of acute kidney injury (AKI) following pancreaticoduodenectomy and related risk factors in elderly patients.Methods:The clinical data of elderly patients who underwent pancreaticoduodenectomy in Henan Provincial People′s Hospital from January 2017 to June 2020 were collected retrospectively. According to the changes of serum creatinine within 48 h or 7 days after operation, the patients were divided into AKI group and non-AKI group. The basic clinical characteristics of the two groups were compared, and the incidence of AKI was calculated. Multivariate logistic regression model was used to analyze the risk factors of postoperative AKI.Results:A total of 322 elderly patients were enrolled, with age of (67.1±5.2) years old (60-85 years old) and 186 males (57.76%). Among 322 elderly patients, there were 41 patients (12.73%) suffering from AKI following pancreaticoduodenectomy. Compared with the non-AKI group, the level of bilirubin in AKI group was higher ( Z=-2.012, P=0.044), and the level of hemoglobin in AKI group was lower ( Z=-2.111, P=0.035). Multivariate logistic regression analysis showed that increased preoperative total bilirubin ( OR=1.003, 95% CI 1.000-1.006, P=0.027) and postoperative exploratory laparotomy ( OR=3.936, 95% CI 1.071-14.460, P=0.039) were the independent influencing factors for AKI after pancreaticoduodenectomy in elderly patients. Conclusions:The incidence of AKI after pancreaticoduodenectomy in elderly patients is 12.73%. Preoperative high bilirubin and postoperative exploratory laparotomy may be the independent risk factors for AKI after pancreaticoduodenectomy in elderly patients.

Objective·To investigate the effect of F-box only protein 38(FBXO38)on the ocular melanoma proliferation and the potential regulatory pathway.Methods·Human skin cutaneous melanoma A375 and human uveal melanoma OMM2.3 cell lines with FBXO38 knockdown and overexpression were constructed by FBXO38 short hairpin RNA(shRNA)and FBXO38 overexpression plasmids respectively.Knockdown and overexpression efficiency of FBXO38 at transcription and protein levels were verified by using quantitative real-time PCR(qRT-PCR)and Western blotting.The effects of FBXO38 on melanoma cell proliferation were detected through clonal formation assay,BrdU immunofluorescence staining and CCK8 cell proliferation assay.By using The Cancer Genome Atlas(TCGA)database,differentially expressed genes were analyzed in the high and low expression groups of FBXO38.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment was performed to reveal the signaling pathways associated with FBXO38.CCK8 cell proliferation assay was used to detect the inhibition rates of the signaling pathway inhibitors on cells with different FBXO38 expression levels.qRT-PCR and Western blotting were used to detect whether the signaling pathway was activated after knocking down FBXO38.Results·qRT-PCR and Western blotting verified that mRNA and protein expression levels of FBXO38 in FBXO38 knockdown A375 and OMM2.3 cell lines decreased compared with the control group,while the expression levels of FBXO38 in the overexpression cell lines increased compared with wild type group(P＜0.05).Clonal formation assay,BrdU immunofluorescence staining and CCK8 cell proliferation assay showed that FBXO38 knockdown significantly enhanced the proliferation of A375 and OMM2.3 cells(P＜0.05),while overexpression of FBXO38 inhibited melanoma cell proliferation(P＜0.05).Enrichment analysis showed that in skin cutaneous melanoma and uveal melanoma,FBXO38 expression influenced the phosphoinositide 3-kinase/protein kinase B(PI3K-Akt)pathway activation.Compared with those in the control group,the inhibition rates of P13K inhibitor LY294002 and mTOR1 inhibitor Everolimus in the FBXO38 knockdown group significantly improved(P＜0.05),while their inhibition rates of the overexpression group significantly decreased compared with those of control cells(P＜0.05).Western blotting results showed that after knocking down FBXO38,expression levels of PTEN,P21 and P53 proteins decreased,while expression level of MDM2 protein increased.The qRT-PCR results showed a significant decrease in P53 transcription level(P＜0.05)and a significant increase in MDM2 transcription level in FBXO38 knockdown cells(P＜0.05).Conclusion·FBXO38 plays a role in regulating the proliferation of ocular melanoma,and this regulatory effect is related to the PI3K-Akt signaling pathway.

Objective To observe the influence of the staining phenomenon after fluorescein sodium staining on eye irritation in normal rabbits.MethodsIn the experimental rabbit eye irritation test conducted with sodium chloride eye drops, Siwei Zhenceng Bingpeng eye drops, sodium hyaluronate eye drops, sodium cromoglycate eye drops, and compound aspartate eye drops (4 in each group, half male and half female), the left eyes of rabbits were administered normal saline (self-negative control) and the right eyes were administered the experimental medicine; the eyes were stained with 1% sodium fluorescein, and eye irritation was observed and scored using slit lamp microscope for 31 days. Morphological changes of corneal epithelial staining were recorded and the incidence of staining was calculated. After the observation, the eyeballs and Hasselblad glands were examined histopathologically, and the staining rate of the left eye was compared with that of the right eye which was administered the corresponding medicine.ResultsNeither eye had any irritation symptoms; the scores were 0, and the total incidences of corneal staining were 3% (left) and 1% (right), respectively. There was no significant difference between the two groups (P > 0.05). Corneal epithelial staining showed single-spot staining, scattered dot, localized, or large areas of fusion staining. No histopathological changes were found in the eyeballs or Hasselblad glands, and the results were evaluated as non-irritative.Conclusion The irregularity of corneal epithelial staining in rabbits did not influence the results of the ocular irritation test.

Objective: To evaluate the risk of 10-year risk of ischemic cardiovascular disease (ICVD) in patients with type 2 diabetes aged 35 years old and above in two cities of Jiangsu province. Methods: From December 2013 to January 2014, a total of 15 624 patients with type 2 diabetes aged 35 years old and above, who received national basic public health service in Changshu county of Suzhou city, Huai′an and Qinghe districts of Huai′an city, Jiangsu province, were recruited by cluster sampling method. Face-to-face questionnaire survey, anthropometric and laboratory measurements were conducted to collect exposures to ICVD risk factors. Improved Ten Year Risk Assessment Table of ICVD in Chinese was used to assess the risk score and the absolute risk of developing ICVD. Results: The participants were (62.2±9.7) years old, of which 6 137 were men (39.3%). Among the participants, the highest rate of exposure to ICVD risk factors was high systolic blood pressure (74.8%, n=11 685), followed by high total cholesterol (70.7%, n= 11 051).The score of 10-year risk for ICVD was (10.4±3.3) and the median (P₂₅-P₇₅) value of absolute risk was 15.6% (6.8%-32.7%). 16.7% (n=2 602) participants were under extremely high risk of 10-year risk for ICVD, 23.8% (n=3 714) under high-risk and 24.0% (n=3 746) under middle-risk. Among the total risk score of ICVD, age (49.1%), hypertension (17.7%) and diabetes (15.5%) accounted for relatively high proportion, however, smoking (11.0%) was the most important risk factor except for age (47.4%) and systolic blood pressure (20.5%) in male participants. Conclusion Patients with type 2 diabetes aged 35 years old and above in two cities of Jiangsu Province have a high risk of developing ICVD for 10 years, especially in elderly, female, hypertension patients and male smokers.

Objective: To investigate the relationship between duration of sleep and all-cause mortality in patients with type 2 diabetes. Methods: A total of 17 452 type 2 diabetic patients who were under the National Basic Public Health Service Project, in Changshu county, Qinghe district and Huai’an district in Huai’an city of Jiangsu province, were recruited as participants. Cox proportional hazards regression models were used to estimate the associations between different levels on the duration of sleep and all-cause mortality. Stratified analysis was performed according to factors as gender, age, and lifestyle. Results: Among the 67 912 person-years of observation, from the fo1low- up program, the median time of follow-up was 4 years, with 1 057 deaths occurred during the follow-up period. Taking patients with duration of sleep as 7 h/d for reference, the multivariate adjusted hazard ratios (95%CI) of all-cause mortality appeared as ≤6, 8, 9 and ≥10 h/d were 1.14 (0.94-1.37), 1.10 (0.91-1.32), 1.33 (1.05-1.70), and 1.52 (1.24-1.87), respectively which were associated with the duration of sleep. Data from the subgroup analysis showed that this was consistent with the whole population. Longer duration of sleep was associated with the all-cause mortality but no significant association was found between shorter duration of sleep and the all-cause mortality. Conclusion Longer duration of sleep seemed to have increased the risk of all-cause mortality in type 2 diabetic patients.

Modulating Tankyrases (TNKS), interactions with USP25 to promote TNKS degradation, rather than inhibiting their enzymatic activities, is emerging as an alternative/specific approach to inhibit the Wnt/β-catenin pathway. Here, we identified UAT-B, a novel neoantimycin analog isolated from Streptomyces conglobatus, as a small-molecule inhibitor of TNKS-USP25 protein-protein interaction (PPI) to overcome multi-drug resistance in colorectal cancer (CRC). The disruption of TNKS-USP25 complex formation by UAT-B led to a significant decrease in TNKS levels, triggering cell apoptosis through modulation of the Wnt/β-catenin pathway. Importantly, UAT-B successfully inhibited the CRC cells growth that harbored high TNKS levels, as demonstrated in various in vitro and in vivo studies utilizing cell line-based and patient-derived xenografts, as well as APC^min/+ spontaneous CRC models. Collectively, these findings suggest that targeting the TNKS-USP25 PPI using a small-molecule inhibitor represents a compelling therapeutic strategy for CRC treatment, and UAT-B emerges as a promising candidate for further preclinical and clinical investigations.