Objective:To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the fast determination of maduramicin ammonium (MAD) in rat serum.Methods:In February 2024, rat serum samples were selected and directly injected after extraction and purification with methanol: acetonitrile (1: 1), separated on a C18 chromatographic column, and gradient-eluted using 0.1% formic acid aqueous solution -0.1% formic acid methanol solution as the mobile phase. Under optimized instrument conditions, electrospray positive ion multiple reaction monitoring (MRM) mode was employed for quantification using the external standard method, followed by methodological validation of the established approach.Results:The linearity of MAD in serum was good in the concentration range of 0.5-100 μg/L, and the correlation coefficient was 0.9997. The mean recoveries of MAD from spiked samples were 86.0%-109.6%, with the relative standard deviations were less than 10%. The limit of detection was 0.23 μg/L, The limit of detection was 0.75 μg/L.Conclusion:This method is high sensitive and reliable, which is suitable for the determination of MAD in in mouse serum.

Objective:To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the fast determination of maduramicin ammonium (MAD) in rat serum.Methods:In February 2024, rat serum samples were selected and directly injected after extraction and purification with methanol: acetonitrile (1: 1), separated on a C18 chromatographic column, and gradient-eluted using 0.1% formic acid aqueous solution -0.1% formic acid methanol solution as the mobile phase. Under optimized instrument conditions, electrospray positive ion multiple reaction monitoring (MRM) mode was employed for quantification using the external standard method, followed by methodological validation of the established approach.Results:The linearity of MAD in serum was good in the concentration range of 0.5-100 μg/L, and the correlation coefficient was 0.9997. The mean recoveries of MAD from spiked samples were 86.0%-109.6%, with the relative standard deviations were less than 10%. The limit of detection was 0.23 μg/L, The limit of detection was 0.75 μg/L.Conclusion:This method is high sensitive and reliable, which is suitable for the determination of MAD in in mouse serum.

Objective:To evaluate the effect of robot-assisted laparoscopic technology via abdominal approach for patients with primary retroperitoneal tumors.Methods:A retrospective cohort analysis was conducted for the clinical data of 71 patients who underwent robot-assisted laparoscopic resection of primary retroperitoneal tumor via abdominal approach at the Department of Urology of Sir Run Run Shaw Hospital,Zhejiang University School of Medicine from January 2015 to December 2023. There were 35 male and 36 female patients. The age ( M(IQR)) was 56(21) years (range: 21 to 83 years). The median tumor diameter was 46 (31) mm (range: 15 to 134 mm). Postoperative pathology revealed 58 benign and 13 malignant cases. Patients were divided into non-adherent group ( n=47) and adherent group ( n=24) based on whether the tumor was adhered to major organs or vessels. Perioperative and postoperative situation were compared between the two groups. Data comparisons were conducted using independent samples t-test for normally distributed continuous variables, Mann-Whitney U tests for non-normally distributed data, χ2 test or Fisher′s exact test for categorical variables. Kaplan-Meier survival analysis was employed to estimate 3-year recurrence or metastasis rate and 3-year mortality rate. Results:Operative time was 120(60) minutes (range: 45 to 440 minutes), intraoperative blood loss was 50 (80) ml (range: 10 to 2 000 ml). The median change of intraoperative mean arterial pressure was 40 (19) mmHg(1 mmHg=0.133 kPa)(range: 10 to 112 mmHg). Intraoperative blood transfusion was required in 7 cases, whereas 64 cases did not necessitate transfusion. The change in hemoglobin levels before and after surgery was (17.9±13.6) g/L (range:-16 to 53 g/L), and the median change in serum creatinine levels was 2.0 (14.5) μmol/L (range:-71.0 to 100.4 μmol/L). Postoperative fasting duration was 2.0 (1.5) days (range: 1 to 6 days), and the median hospital stay was 10.0 (7.5) days (range: 4 to 24 days). No perioperative mortality occurred in any of the patients. The non-adherent group had shorter operation time, less estimated blood loss, lower blood transfusion rate, smaller delta value of hemoglobin before and after surgery, larger delta value of creatinine before and after surgery, fewer postoperative complications, shorter postoperative fasting time, and shorter length of hospital stay than the adherent group(all P<0.05), while there was no significant difference in mean arterial pressure fluctuation between the two groups ( P>0.05). Follow-up data were available for 69 patients, with a median follow-up duration of 39 (43) months (range: 4 to 88 months). Among these patients, 40 completed the 3-year follow-up. The 3-year recurrence or metastasis rate was 10.14%, and the 3-year mortality rate was 2.90%. Conclusions:Robot-assisted laparoscopic technology via abdominal approach for resection of primary retroperitoneal tumors is safe and feasible. It can also achieve secure surgical outcome for primary retroperitoneal tumors adherent to surrounding organs or vessels, albeit with increased surgical complexity and slower postoperative recovery compared to non-adherent cases.

Objective:To evaluate the effect of robot-assisted laparoscopic technology via abdominal approach for patients with primary retroperitoneal tumors.Methods:A retrospective cohort analysis was conducted for the clinical data of 71 patients who underwent robot-assisted laparoscopic resection of primary retroperitoneal tumor via abdominal approach at the Department of Urology of Sir Run Run Shaw Hospital,Zhejiang University School of Medicine from January 2015 to December 2023. There were 35 male and 36 female patients. The age ( M(IQR)) was 56(21) years (range: 21 to 83 years). The median tumor diameter was 46 (31) mm (range: 15 to 134 mm). Postoperative pathology revealed 58 benign and 13 malignant cases. Patients were divided into non-adherent group ( n=47) and adherent group ( n=24) based on whether the tumor was adhered to major organs or vessels. Perioperative and postoperative situation were compared between the two groups. Data comparisons were conducted using independent samples t-test for normally distributed continuous variables, Mann-Whitney U tests for non-normally distributed data, χ2 test or Fisher′s exact test for categorical variables. Kaplan-Meier survival analysis was employed to estimate 3-year recurrence or metastasis rate and 3-year mortality rate. Results:Operative time was 120(60) minutes (range: 45 to 440 minutes), intraoperative blood loss was 50 (80) ml (range: 10 to 2 000 ml). The median change of intraoperative mean arterial pressure was 40 (19) mmHg(1 mmHg=0.133 kPa)(range: 10 to 112 mmHg). Intraoperative blood transfusion was required in 7 cases, whereas 64 cases did not necessitate transfusion. The change in hemoglobin levels before and after surgery was (17.9±13.6) g/L (range:-16 to 53 g/L), and the median change in serum creatinine levels was 2.0 (14.5) μmol/L (range:-71.0 to 100.4 μmol/L). Postoperative fasting duration was 2.0 (1.5) days (range: 1 to 6 days), and the median hospital stay was 10.0 (7.5) days (range: 4 to 24 days). No perioperative mortality occurred in any of the patients. The non-adherent group had shorter operation time, less estimated blood loss, lower blood transfusion rate, smaller delta value of hemoglobin before and after surgery, larger delta value of creatinine before and after surgery, fewer postoperative complications, shorter postoperative fasting time, and shorter length of hospital stay than the adherent group(all P<0.05), while there was no significant difference in mean arterial pressure fluctuation between the two groups ( P>0.05). Follow-up data were available for 69 patients, with a median follow-up duration of 39 (43) months (range: 4 to 88 months). Among these patients, 40 completed the 3-year follow-up. The 3-year recurrence or metastasis rate was 10.14%, and the 3-year mortality rate was 2.90%. Conclusions:Robot-assisted laparoscopic technology via abdominal approach for resection of primary retroperitoneal tumors is safe and feasible. It can also achieve secure surgical outcome for primary retroperitoneal tumors adherent to surrounding organs or vessels, albeit with increased surgical complexity and slower postoperative recovery compared to non-adherent cases.

Background Individuals may experience significant alterations in sleep hygiene during the major public health emergencies,consequently impacting their sleep quality.Objective To explore the relationship between sleep quality and sleep hygiene among adult residents during the major public health emergencies,so as to provide references for improving the sleep quality of residents during such a period.Methods A sample of 1 364 adult residents were enrolled as the research subjects from February 20 to 29,2020.All participants were asked to complete self-administered questionnaire to obtain basic-demographic information and sleep hygiene.Pittsburgh Sleep Quality Index(PSQI)was applied to assess sleep quality.Residents were classified into poor sleepers with PSQI score≥8 and good sleepers defined as PSQI score<8.Binary Logistic regression analysis was conducted to identify factors associated with sleep quality.Radar chart was used to visualize and compare the sleep hygiene between poor sleepers and good sleepers.Results According to PSQI score,891(65.32%)residents were good sleepers,while 473(34.68%)residents were poor sleepers.Comparison revealed that age(χ2=3.887),past medical history(χ2=27.938),awareness rate of importance of sleeping before major public health emergencies(χ2=4.337),impact of sleep quality on quality of life during the major public health emergencies(χ2=178.138),frequency of staying up late during the major public health emergencies(χ2=139.390),compensatory sleep behaviors(χ2=39.257),impact of sleep problems on daytime functioning(χ2=285.879),change of bedtime(χ2=63.031),sleep latency(χ2=168.672),wake-up time(χ2=59.221),changes in sleep duration(χ2=172.332),time spent in the bedroom(χ2=23.071),and sum of money spent on improving sleep environment(χ2=58.584)yielded statistical difference between poor sleepers and good sleepers(P<0.05 or 0.01).Logistic regression analysis denoted that past medical history(OR=1.680,95%CI:1.185～2.382),negative impact of sleep quality on quality life(OR=4.181,95%CI:2.722～6.422),staying up late 3 to 4 times per week(OR=3.145,95%CI:1.497～6.605),staying up late almost every day(OR=4.271,95%CI:1.970～9.260),negative impact of sleep problems on daytime functioning(OR=7.169,95%CI:5.188～9.907),prolonged sleep latency(OR=2.836,95%CI:2.019～3.982)and shortened sleep duration(OR=3.518,95%CI:2.144～5.772)were risk factors of poor sleep quality.The sum of money spent on improving sleep environment following the major public health emergencies≤500 RMB(OR=0.334,95%CI:0.134～0.830)was related to the incidence rate of poor sleep quality.Radar chart showed that poor sleepers were characterized by extravagant concerns,excessive cleanliness and poor sleep hygiene practices during the major public health emergencies,and poor sleepers were more likely to stay up late due to stress and emotional issues.Conclusion Some residents are facing poor sleep quality during the major public health emergencies,and poor sleep hygiene practice also contributes to poor sleep quality.

【Objective】 M3 muscarinic acetylcholine receptor(M3 receptor), encoded by CHRM3 gene, is widely distributed in the cardiovascular system and plays an important role in cardiac regulation. The aim of this study was to assess the association of genetic variants in M3 receptor with blood pressure(BP) responses to controlled dietary sodium and potassium interventions. 【Methods】 A total of 333 subjects from 124 families were recruited from the rural areas of northern China. After a three-day baseline observation, they were sequentially on a seven-day low-salt diet, a seven-day high-salt diet, and a seven-day high-salt diet plus potassium supplementation. Thirteen CHRM3 single nucleotide polymorphisms(SNPs) were selected for analysis. 【Results】 SNP rs10802811 of the CHRM3 was significantly associated with diastolic BP(DBP) and mean arterial pressure(MAP) responses to both low-salt and high-salt diets while SNPs rs6429147, rs373288072, rs114677844 and rs663148 showed significant associations with systolic BP(SBP) and MAP responses to high-salt diet. In addition, SNP rs6692904 was significantly associated with SBP, DBP and MAP responses to high-salt diet with potassium supplementation. 【Conclusion】 Genetic variants in M3 receptor are significantly associated with BP responses to sodium and potassium intervention, suggesting that M3 receptor may be mechanistically involved in BP salt and potassium sensitivity.

Abstract： Objective　To elucidate the antigenic antibody reaction of recombinant expression of non-structural protein 1 (NS1) of Japanese encephalitis (JE) virus with various mosquito-borne flaviviruses, including JE virus, and the antigenic antibody reaction of serum samples of patients infected with JE virus in acute stage. Methods　In this study, Escherichia coli prokaryotic expression vector (pET) system was used to recombinant express Japanese encephalitis virus NS1 gene. Western Blot assay was performed to detect the antibody responses of the recombinantly expressed protein against a variety of mosquito-transmitted flaviviruses, including JE virus, as well as antigen-antibody reactions of serum from patients with acute JE virus infection. Results　The NS1 gene expression product of JE virus (P3 strain) was in the form of an inclusion body, and the denatured and renatured expression product was displayed as a single band in the denatured gel (polyacrylamide gel electrophoresis, PAGE), with a molecular weight of about 45 000. The results of further antigen-antibody analysis showed that the antigen/antibody hybridization reaction of the expression product with polyclonal or monoclonal antibody of JE virus (mosquito isolates, encephalitis isolates) and serum samples of patients with acute JE virus infection could be completely consistent. The recombinant product showed negative antigen/antibody hybridization reactions with mosquito-transmitted flaviviruses, such as dengue virus and yellow fever virus polyclonal antibodies, but positive reactions with polyclonal antibodies to West Nile virus and Murray Valley encephalitis virus. Conclusions　In this study, the recombinant expression of the NS1 protein of JE virus was successfully obtained, and the antigen/antibody reaction between the recombinant protein and samples of patients infected with mosquito-borne flavivirus and JE virus was analyzed. The study results provide important basic data for elucidating the antigen-antibody reaction between the NS1 protein of JE virus and mosquito-borne flavivirus. The recombinant expression protein obtained in this study provides an important material basis for further research on the function of JE virus NS1 protein.

【Objective】 To explore the effects of dietary salt intake on serum and urinary levels through the chronic salt loading intervention. 【Methods】 Eighty adults （18 to 65 years old） were screened from two villages in Liquan and Lantian counties to participate in a 2-week chronic salt intervention， including a 3-day baseline survey， a 7-day low-salt diet， and a 7-day high-salt diet. Uromodulin levels in serum and urine were determined by enzyme-linked immunosorbent assay （ELISA） kits. According to the baseline blood pressure levels， all subjects were divided into normotensive and hypertensive groups. Pearson or Spearman correlation analyzed the associations of 24 h urinary sodium excretions with serum and urinary levels of uromodulin. 【Results】 At the baseline， serum uromodulin in hypertensive subjects was significantly lower than that in normotensive subjects （26.7±9.9 vs. 57.9±9.7 ng/mL， P=0.033）. Serum uromodulin levels were significantly lower on a high-salt diet than on a baseline diet ［（54.9±8.8 vs. 28.3±4.5） ng/mL， P=0.007］. In addition， daily urinary excretions of uromodulin were lower on a high-salt diet ［（28.4±6.6） ng/mL］ than on a baseline diet ［（282.1±70.0） ng/mL］ and on a low-salt diet ［（154.1±21.3） ng/mL］. The 24 h urinary sodium excretions were inversely correlated with urinary uromodulin excretions （r=-0.40， P<0.001） on both low-salt and high-salt diets， but not correlated with serum uromodulin levels. 【Conclusion】 Variations in dietary salt intake significantly affect plasma and urine uromodulin levels.

Objective:To investigate the association between body mass index (BMI) trajectories in children and adolescents and subclinical renal damage (SRD) in adulthood.Methods:4 623 participants aged 6-18 years old were recruited from the ongoing cohort of Hanzhong adolescent hypertension study in 1987, and the subjects were followed up in 1989, 1992, 1995, 2005, 2013 and 2017, respectively. Group-based trajectory modeling was used to identify distinct BMI trajectories in longitudinal analysis. Generalized linear model was applied to examine the association between different BMI trajectories and SRD incidence in adulthood.Results:A total of 2 678 subjects from childhood to adulthood were enrolled in this study. All subjects were divided into three groups according to three distinct BMI trajectories: low-increasing BMI group ( n=1 017), moderate-increasing BMI group ( n=1 353), and high-increasing BMI group ( n=308). Over follow up for 30 years, a total of 248 participants (9.3%) developed SRD. Urinary albumin-to-creatinine ratio (uACR) in low to high-increasing BMI group was 0.9(0.6, 1.4), 1.0(0.7, 1.7), 1.6(0.8, 3.2), respectively ( P trend<0.001), and estimated glomerular filtration rate was 98.5(87.6, 111.6) , 96.2(86.4, 109.7), 95.3 (87.5, 125.0) ml·min -1·(1.73 m 2) -1, respectively ( P trend=0.025). The generalized linear model analysis showed that uACR was increased linearly from low to high-increasing BMI group [ β=3.16(95% CI 1.02-5.31), Ptrend=0.004]. There was no correlation or linear trend between BMI trajectory and estimated glomerular filtration rate [ β=-2.30(95% CI-5.18-0.57), Ptrend=0.117]. Compared with the low-increasing BMI group, the high-increasing BMI group had greater odds of experiencing SRD in adulthood after adjusting for multiple confounders such as age, gender, medical history and lifestyle ( OR=2.83, 95% CI 1.84-4.36, Ptrend<0.001). Conclusions:Higher BMI trajectorie is correlated with higher level of uACR and risk of SRD in middle age. Identifying long-term BMI trajectorie from early age may assist in predicting individuals′ renal function in later life.

【Objective】 Based on our previously established salt-sensitive hypertension cohort, we conducted chronic salt loading and potassium supplementation interventions, aiming to examine the association between genetic variants in renalase and blood pressure (BP) responses to dietary interventions of salt and potassium intake. 【Methods】 In 2004, 514 subjects from 126 families were recruited in Shaanxi Province to establish the salt-sensitive hypertension study cohort. Among them, 334 non-parent subjects were selected and sequentially maintained on a low-salt diet for 7 days, then a high-salt diet for 7 days and a high-salt diet with potassium supplementation for another 7 days. Ten single nucleotide polymorphisms (SNPs) in the renalase gene were genotyped on the MassARRAY platform. 【Results】 SNP rs2576178 of the renalasegene was significantly associated with systolic BP (SBP) and mean arterial pressure (MAP) responses to low-salt intervention (SBP: β=-2.730, P<0.05; MAP: β=-1.718, P<0.05). In addition, SNP rs12356177 was significantly associated with diastolic BP response to low-salt diet (β=-1.608, P<0.05). However, we did not find any association for the renalase SNPs with BP response to high-salt diet with potassium supplementation reached nominal statistical significance. 【Conclusion】 Genetic variants in renalase gene are significantly associated with BP response to low-salt diet, suggesting that renalase may be mechanistically involved in BP salt-sensitivity.