Objective:To establish a method for determination of two derivatives of cyanide in biomaterials by headspace gas chromatography mass spectrometry.Methods:In February 2024, blood and urine samples were placed in headspace sampling vials. Phosphoric acid, or phosphoric acid and chloramine T solution, was added respectively to derivatize cyanide into hydrogen cyanide or cyanogen chloride. After equilibrating at 80°C for 15 minutes, headspace sampling was performed. The samples were separated by a GS-GasPro chromatographic column and determined by gas chromatography-mass spectrometry in selected ion monitoring (SIM) mode. Qualitative analysis was conducted using both retention time and the ratio of characteristic ions, and quantitative analysis was carried out by the external standard method.Results:The correlation coefficients (r) for the two derivatives was 0.9992 and 0.9984, respectively. The detection limits was 0.10 μg/ml and 0.05 μg/ml, respectively. The recoveries ranged from 73.7% to 98.4%, with precision (RSD) ranging from 3.3% to 14.2%.Conclusion:The method offers the advantages of simple operation, precise quantification, high qualitative reliability, and high sensitivity, making it suitable for cyanide detection in biological materials.

Objective:To establish a method for determination of two derivatives of cyanide in biomaterials by headspace gas chromatography mass spectrometry.Methods:In February 2024, blood and urine samples were placed in headspace sampling vials. Phosphoric acid, or phosphoric acid and chloramine T solution, was added respectively to derivatize cyanide into hydrogen cyanide or cyanogen chloride. After equilibrating at 80°C for 15 minutes, headspace sampling was performed. The samples were separated by a GS-GasPro chromatographic column and determined by gas chromatography-mass spectrometry in selected ion monitoring (SIM) mode. Qualitative analysis was conducted using both retention time and the ratio of characteristic ions, and quantitative analysis was carried out by the external standard method.Results:The correlation coefficients (r) for the two derivatives was 0.9992 and 0.9984, respectively. The detection limits was 0.10 μg/ml and 0.05 μg/ml, respectively. The recoveries ranged from 73.7% to 98.4%, with precision (RSD) ranging from 3.3% to 14.2%.Conclusion:The method offers the advantages of simple operation, precise quantification, high qualitative reliability, and high sensitivity, making it suitable for cyanide detection in biological materials.

Objective: To construct a human G protein-coupled receptor kinase 2 ( GRK2) eukaryotic expression system． Methods: The primers were designed ，and the His-GRK2 target gene was amplified by PCR using the Pans-EGFP-GrK2 (full-length) gene as the template．The His-GRK2 target gene was connected to the pcDNA3．1EGFP eukaryotic expression vector． The pcDNA3. 1-EGFP-His-GRK2 plasmid was transfected into HEK 293T cells．48 h later，the expression of GRK2 protein was detected by Western blot，and the GRK2 protein was purified by nickel chelated magnetic bead method．The purification of GRK2 protein was detected by Coomassie bright blue staining and Western blot，and the activity of GRK2 protein was detected by His pull down. Results : The results of double enzyme digestion and sequencing showed that pcDNA3. 1-EGFP-His-GRK2 eukaryotic expression plasmid was successfully constructed．Western blot analysis showed that the molecular weight of GRK2 protein was about 80 ku，indicating that GRK2 protein was successfully expressed in HEK 293T cells (t = 6. 433，P = 0. 003) ．GRK2 protein was purified by nickel chelated magnetic beads．His pull down experiment results showed that GRK2 was bound to prostaglandin E2 receptor 4 (EP4) ，suggesting that GRK2 protein had biological activity (t = 13. 5，P = 0. 000 2) ． Conclusion The pcDNA3．1-EGFP-His-GRK2 eukaryotic expression plasmid was correctly sequenced and the GRK2 recombinant plasmid was successfully constructed．The GRK2 recombinant plasmid was successfully expressed in eukaryotic cells HEK 293T and the protein expressed was biologically active.

Objective:To analyze the incidence and epidemiological characteristics of traumatic spinal cord injury in China in 2018.Methods:Multi-stage stratified cluster sampling was used to randomly select hospitals capable of treating patients with spinal cord injury from 3 regions，9 provinces and 27 cities in China to retrospectively investigate eligible patients with traumatic spinal cord injury admitted in 2018. National and regional incidence rates were calculated. The data of cause of injury，injury level，severity of injury，segment and type of fracture，complications，death and other data were collected by medical record questionnaire，and analyzed according to geographical region，age and gender.Results:Medical records of 4，134 patients were included in this study，with a male-to-female ratio of 2.99∶1. The incidence of traumatic spinal cord injury in China in 2018 was 50.484 / 1 million （95% CI 50.122-50.846）. The highest incidence in the Eastern region was 53.791 / 1 million （95% CI 53.217-54.365）. In the whole country，the main causes of injury were high falls （29.58%），as well as in the Western region （40.68%），while the main causes of injury in the Eastern and Central regions were traffic injuries （31.22%，30.10%）. The main injury level was cervical spinal cord in the whole country （64.49%），and the proportion of cervical spinal cord injury in the Central region was the highest （74.68%），and the proportion of lumbosacral spinal cord injury in the Western region was the highest （32.30%）. The highest proportion of degree of injury was incomplete quadriplegia （55.20%），and the distribution pattern was the same in each region. A total of 65.87% of the patients were complicated with fracture or dislocation，77.95% in the Western region and only 54.77% in the Central region. In the whole country，the head was the main combined injury （37.87%），as well as in the Eastern and Central regions，while the proportion of chest combined injury in the Western region was the highest （38.57%）. A total of 32.90% of the patients were complicated with respiratory complications. There were 23 patients （0.56%） died in hospital，of which 17（73.91%） died of respiratory dysfunction. Conclusions:The Eastern region of China has a high incidence of traumatic spinal cord injury. Other epidemiological features include high fall as the main cause of injury cervical spinal cord injury as the main injury level，incomplete quadriplegia as the main degree of injury，head as the main combined injury，and respiratory complications as the main complication.

OBJECTIVE：To e xplore the health economic burden of hypertension patients at county-level areas and its influential factors in China. METHODS ：A questionnaire survey was conducted on hypertension patients in 7 county-level public hospitals from 6 provinces as Hebei ，Shandong，Shanxi provinces by using a convenient sampling method. Catastrophic health expenditure was defined by the standard of “medical and health expenditure exceeding 10％ of household income ”. The incidence ， average gap and relative gap of catastrophic health expenditure were analyzed. A multi-factor Logistic regression model analysis was used to analyze the influential factors that lead to catastrophi c health expenditure. RESULTS ：A total of 1 378 questionnaires were sent out ，and 925 valid questionnaires were collected with effective rate of 67.13％ . The incidence of catastrophic health expenditure，average gap and relative gap among hypertension patients were 23.03％，19.37％ and 84.12％，respectively. At different income levels ，the incidence of catastrophic health expenditure，average gap and relative gap were 72.67％ ， 96.79％ and 133.18％ in the poorest household group ，and were 1.94％ ，0.47％ and 24.23％ in the richest household group. Among different types of medical insurance ，the incidence of catastrophic health expenditure in patients covered by “New Rural Cooperative Medical Scheme （NRCMS）”the highest （31.30％）. The household income ，complications and the type of health insurance had significant impacts on the incidence of catastrophic health expenditure in hypertension patients （P＜0.05）. CONCLUSIONS：The incidence of catastrophic health expenditure in hypertension patients with different income levels is different. As the income level raised ，the incidence of catastrophic health expenditures continued to decrease. But the protection of household health expenditure by NRCMS is weak. It is suggested that a certain policy preference should be given to families with low income and patients with chronic diseases ，so as to ensure the rights and interests of patients with hypertension .

OBJECTIVE： To evaluate the economics of the full coverage policy for hypoglycemic essential medicines in Taizhou city， and to provide reference for improving medicine accessibility and essential medicine system in China. METHODS： The electronic health records and financial input data of diabetic patients were extracted from different districts and counties of Taizhou city during 2009-2016. In cohort study design， taking the implementation time of each district and county’s policies as the breakpoint， the data recorded annually were processed as annual data， forming the cohort data for 1 year before （baseline year） and 3 years after the implementation of the policies. According to the choice of free medicines after the implementation of the policy， they were divided into policy group and control group. Propensity score matching was used to balance the differences between two groups to get the final sample， and then the economics of the policy was evaluated with the methods of difference-in-differences. The cost and benefit of implementation policy were calculated. RESULTS： Totally 14 744 people of each group were got by propensity score matching. The mean annual financial expenses on free medicine per capita were 263.8 yuan. Compared with baseline year， mean annual medicine expenses per capita reduced by the policy were 649.2 yuan， and mean annual hospitalization expenses per capita were 624.7 yuan. Thus the input-output ratio of the policy was 1 ∶ 4.8. CONCLUSIONS： The implementation of the policy is beneficial to reduce the cost burden of patients and economical. It can be popularized and sustained through scientific design combined with relevant policies.

Pigment epithelium-derived factor (PEDF) is an antiangiogenic factor which is effective in tumour inhibition in a variety of tumours and has not yet been studied in bladder tumour before. In this study the expression of PEDF, interleukin-1α (IL-1α) and -8 (IL-8) in bladder tumours was investigated. Immunohistochemistry was performed on 64 bladder tumour and 23 normal uroepithelium samples. Expression change of the factors was compared with clinicopathological parameters. Correlations between PEDF, IL-1α and IL-8 were analyzed. None of the factors was in relation to gender, tumour occurrence, and size or onset pattern. PEDF (P=0.014) and IL-1α (P=0.049) expression was down-regulated with grade progression. PEDF expression was lower in normal uroepithelium than in papillary urothelial neoplasm of low malignant potential (PUNLMP) (P=0.000) and carcinoma (P=0.009) whilst IL-1α (P=0.000 and P=0.000 respectively) and IL-8 (P=0.000 and P=0.023 respectively) expression was higher in the same grouping. PEDF expression had a negative correlation with IL-8 in PUNLMP (P=0.049, r=-0.578) as well as in tumour grouping (P=0.033, r=-0.276). Deranged expressional change of PEDF, IL-1α and IL-8 could be in relation to loss of differentiation from normal uroepithelium to papillary lesion and eventually to carcinoma.

Pigment epithelium-derived factor (PEDF) is an antiangiogenic factor which is effective in tumour inhibition in a variety of tumours and has not yet been studied in bladder tumour before.In this study the expression of PEDF,interleukin-1 α (IL-1α) and -8 (IL-8) in bladder tumours was investigated.Immunohistochemistry was performed on 64 bladder tumour and 23 normal uroepithelium samples.Expression change of the factors was compared with clinicopathological parameters.Correlations between PEDF,IL-1α and IL-8 were analyzed.None of the factors was in relation to gender,tumour occurrence,and size or onset pattem.PEDF (P=0.014) and IL-1α (P=0.049) expression was down-regulated with grade progression.PEDF expression was lower in normal uroepithelium than in papillary urothelial neoplasm of low malignant potential (PUNLMP) (P=0.000) and carcinoma (P=0.009) whilst IL-1α (P=0.000 and P=0.000 respectively) and IL-8 (P=0.000 and P=0.023 respectively) expression was higher in the same grouping.PEDF expression had a negative correlation with IL-8 in PUNLMP (P=0.049,r=-0.578) as well as in tumour grouping (P=0.033,r=-0.276).Deranged expressional change of PEDF,IL-1 α and IL-8 could be in relation to loss of differentiation from normal uroepithelium to papillary lesion and eventually to carcinoma.