Human tissue kallikrein-binding protein (Kallistatin, KAL), a secretory protein that participates in the regulation of multiple signaling pathways by binding to the extracellular receptor, however, at present has not been reported about the intracellular activity, and whether it has the similar biological activity with extracellular activity. Here we constructed no signal peptide KAL (NSK) into the adeno-associated virus vector to explore the intracellular activity of KAL. Both the endothelial cell and lung cancer cells could express KAL, but not secreted after rAAV2-NSK transfection. The proliferation and migration of human umbilical vein endothelial cells (HUVECs) were inhibited, but the apoptosis rate was not affected. The proliferation rates, mobility and tubule formation of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited to different extents. This cellular study not only confirmed the intracellular activity, but also suggested it may serve as a kind of "balance factor" in multi-targeted controlling, which may provide a new train of thoughts to explain the regulatory contradiction in PI3K-Akt signaling pathways by KAL.
Apoptosis ; Cell Proliferation ; Dependovirus ; Genetic Vectors ; Human Umbilical Vein Endothelial Cells ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; Serpins ; metabolism ; Signal Transduction ; Transfection

A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine butyrate and its primary metabolite clevidipine acid in dog blood. After one-step protein precipitation with methanol, the chromatographic separation was carried out on an Ecosil C18 column (150 mm x 4.6 mm, 5 µm) with a gradient mobile phase consisting of methanol and 5 mmol · L(-1) ammonium formate. A chromatographic total run time of 13.0 min was achieved. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 454.1 [M-H]- --> m/z 234.1 for clevidipine butyrate, m/z 354.0 [M-H]- --> m/z 208.0 for clevidipine acid and m/z 256.1 [M-H]- --> m/z 227.1 for elofesalamide (internal standard, IS) in the negative ion mode with electrospray ionization (ESI) source. The linear calibration curves for clevidipine butyrate and clevidipine acid were obtained in the concentration ranges of 0.5-100 ng · mL and 1-200 ng · mL(-1), separately. The lower limit of quantification of clevidipine butyrate and clevidipine acid were 0.5 ng · mL(-1) and 1 ng · mL(-1). The intra and inter-assay precisions were all below 12.9%, the accuracies were all in standard ranges. Stability testing indicated that clevidipine butyrate and clevidipine acid in dog blood with the addition of denaturant methanol was stable under various processing and/or handling conditions. The validated method has been successfully applied to a pharmacokinetic study of clevidipine butyrate injection to 8 healthy Beagle dogs following intravenous infusion at a flow rate of 5 mg · h(-1) for 0.5 h.
Animals ; Butyrates ; blood ; pharmacokinetics ; Calibration ; Chromatography, Liquid ; Dogs ; Infusions, Intravenous ; Pyridines ; blood ; pharmacokinetics ; Tandem Mass Spectrometry

Objective To investigate the occurrence and reporting of sharp injuries among health care workers (HCWs)at all levels of hospitals in Xuzhou City,provide evidences for formulating protective measures against sharp injuries and improving the reporting system.Methods From July to August 2016,13 hospitals in Xuzhou City were randomly selected by multi-stage stratified random sampling method,the general information,occurrence of sharp injuries,and reporting situation was performed questionnaire survey.Results A total of 2 694 valid question-naires were collected,incidence,case incidence,and reporting rate of sharp injuries were 10.32%,12.84%,and 30.64% respectively.Case incidence of sharp injuries among HCWs in primary,secondary,and tertiary hospitals were 44.83%,11.53%,and 12.52% respectively,case incidences of sharp injuries in different levels of hospitals were significantly different (χ2＝55.148,P＜0.001).The main opportunity for sharp injuries was when HCWs re-turned needle cap (79 cases,22.83%),the main device involving sharp injuries was hollow-bore needle (297 cases, 85.84%).Incidences of sharp injuries among HCWs receiving different training were significantly different (χ2＝66.760,P＜0.001).Conclusion Current situation of sharp injuries among HCWs in this region is not optimistic, there are some problems such as poor training efficacy,low reporting rate and low use rate of safety devices,effec-tive measures should be taken to establish an effective monitoring and tracking system for sharp injuries,so as to re-duce the occurrence of sharp injuries.

Academic Dissertations as Topic ; Epidemiologic Research Design ; Molecular Epidemiology

Objective To develop a knowlodge-attitude-practice questionnaire (KAPQ) for abdominal surgery patients,in order to provide an available tool to evaluate the KAP status of patients' postoperative pain.Methods The KAPQ was established on the basis of clinical experience,literature review and expert consultation.Ninety patients with abdominal surgery in a three level hospital in Jiangsu province were chosen by convenient sampling and SPSS was used for the scale reliability and validity.Results The internal consistency reliabilities of the KAPQ and its three subscales were respectively 0.899,0.933,0.903 and 0.888.The copy of reliability of three subscales were respectively 0.886,0.763 and 0.821.The content coefficients of three subscales were respectively 0.946,0.921 and 0.949.Factor analysis showed that the questionnaire had good construct validity with a common explaining variance more than 0.4.Conclusions The KAPQ has good reliability and validity,thus it can be used as an effective tool to evaluate the health-related knowledge-attitudepractice of the abdominal surgery patients' postoperative pain.

Adipose tissue is a critical energy reservoir in animals and humans, with multifaceted roles in endocrine regulation, immune response, and providing mechanical protection. Based on anatomical location and functional characteristics, adipose tissue can be categorized into distinct types, including white adipose tissue (WAT), brown adipose tissue (BAT), beige adipose tissue, and pink adipose tissue. Traditionally, adipose tissue research has centered on its morphological and functional properties as a whole. However, with the advent of single-cell transcriptomics, a new level of complexity in adipose tissue has been unveiled, showing that even under identical conditions, cells of the same type may exhibit significant variation in morphology, structure, function, and gene expression——phenomena collectively referred to as cellular heterogeneity. Single-cell transcriptomics, including techniques like single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq), enables in-depth analysis of the diversity and heterogeneity of adipocytes at the single-cell level. This high-resolution approach has not only deepened our understanding of adipocyte functionality but also facilitated the discovery of previously unidentified cell types and gene expression patterns that may play key roles in adipose tissue function. This review delves into the latest advances in the application of single-cell transcriptomics in elucidating the heterogeneity and diversity within adipose tissue, highlighting how these findings have redefined the understanding of cell subpopulations within different adipose depots. Moreover, the review explores how single-cell transcriptomic technologies have enabled the study of cellular communication pathways and differentiation trajectories among adipose cell subgroups. By mapping these interactions and differentiation processes, researchers gain insights into how distinct cellular subpopulations coordinate within adipose tissues, which is crucial for maintaining tissue homeostasis and function. Understanding these mechanisms is essential, as dysregulation in adipose cell interactions and differentiation underlies a range of metabolic disorders, including obesity and diabetes mellitus type 2. Furthermore, single-cell transcriptomics holds promising implications for identifying therapeutic targets; by pinpointing specific cell types and gene pathways involved in adipose tissue dysfunction, these technologies pave the way for developing targeted interventions aimed at modulating specific adipose subpopulations. In summary, this review provides a comprehensive analysis of the role of single-cell transcriptomic technologies in uncovering the heterogeneity and functional diversity of adipose tissues.

Objective To evaluate the molluscicidal effect of niclosamidate against Oncomelania hupensis in laboratory and explore its mechanism by determining the enzyme activities of six important enzymes in snail soft tissues.Methods O.hupensis snails were treated with niclosamidate at the concentration of 1.25 mg/L for 24 h and the snail soft tissues were separated and pre-pared for analysis.The enzyme activities of NOS,AChE,SDH,LDH,ACP and AKP were determined by ultraviolet spectropho-tometry.The morphology of the snail soft tissue was also observed.Results Niclosamidate exhibited a potent molluscicidal ef-fect against O.hupensis at the concentration of 5.00 mg/L with a mortality of 96.67% by the immersion method in laboratory.Af-ter immersed with niclosamidate(1.25 mg/L)for 24 h,the enzyme activities of NOS,AChE,ACP and AKP were significantly decreased compared with those of the controls(all P<0.01).There were no significant changes observed in the enzyme activi-ties of SDH and LDH(both P>0.05).Conclusion Niclosamidate possesses a potent molluscicidal effect against O.hupensis and its molluscicidal mechanism is probably by affecting the transmission of neurotransmitters,interfering with the circulation, metabolism and motor functions that require NO,and hindering the digestion and absorption of nutriments,which eventually re-sult in the death of the snails.

BACKGROUNDGaucher's disease (GD) is an autosomal recessive disorder caused by a deficiency of acid β-glucosidase (glucocerebrosidase [GBA]) that results in the accumulation of glucocerebroside within macrophages. Many mutations have been reported to be associated with this disorder. This study aimed to discover more mutations and provide data for the genetic pattern of the gene, which will help the development of quick and accurate genetic diagnostic tools for this disease.

METHODSGenomic DNA was obtained from peripheral blood leukocytes of the patient and Sanger sequencing is used to sequence GBA gene. Sequence alignments of mammalian β-GBA (GCase) and three-dimensional protein structure prediction of the mutation were made. A construct of this mutant and its compound heterozygous counterpart were used to measure GCase in vitro.

RESULTSGCase is relatively conserved at p.T219A. This novel mutation differs from its wild-type in structure. Moreover, it also causes a reduction in GCase enzyme activity.

CONCLUSIONThis novel mutation (c.655A>G, p.T219A) is a pathogenic missense mutation, which contributes to GD.

Child, Preschool ; Gaucher Disease ; genetics ; Glucosylceramidase ; chemistry ; genetics ; Humans ; Male ; Models, Molecular ; Mutation, Missense ; Protein Structure, Tertiary ; Sequence Analysis, DNA

Objective:To investigate the correlation between complement C3 and urine protein level and proteinuria remission status in patients with primary membranous nephropathy (PMN), and better guide individualized clinical treatment.Methods:It was a single-center retrospective study. The clinical data of PMN patients who underwent renal biopsy in the First Affiliated Hospital of Nanjing Medical University from January 2017 to June 2022 were collected. Patients with 24 h urinary protein ≥ 3.5 g were followed up after receiving standard treatment, and the last outpatient or inpatient review was used as the end point of follow-up. 24 h urine protein was collected to evaluate the remission status of proteinuria. Kaplan-Meier method was used to analyze the correlation between serum and renal complements and proteinuria remission. Cox regression analysis method was used to analyze the correlation between serum C3 level and renal tissue C3 deposition and proteinuria remission.Results:This study included 507 PMN patients with 312 (61.54%) males, aged 54 (43, 64) years old. Compared with 24 h urinary protein < 3.5 g group, proportion of males ( χ2=22.479, P<0.001), age ( Z=-2.521, P=0.012), systolic blood pressure ( Z=-4.148, P<0.001), diastolic blood pressure ( Z=-4.084, P<0.001), serum anti-phospholipase A2 receptor (PLA2R) antibody titer ( Z=-7.019, P<0.001), total cholesterol ( Z=-8.796, P<0.001), triglyceride ( Z=-6.158, P<0.001), low density lipoprotein cholesterol ( Z=-8.716, P<0.001), serum creatinine ( Z=-7.368, P<0.001), serum C3 ( Z=-3.663, P<0.001), serum C4 ( Z=-6.560, P<0.001), proportion of glucocorticoid use ( χ2=116.417, P<0.001) and proportion of immunosuppressant use ( χ2=53.839, P<0.001) were all higher, while serum albumin ( Z=12.518, P<0.001), estimated glomerular filtration rate ( Z=6.345, P<0.001) and serum IgG ( Z=7.321, P<0.001) were all lower in 24 h urinary protein ≥3.5 g group. There were 268 patients included in the follow-up cohort with baseline 24 h urinary protein of 7.15 (5.14, 10.24) g, serum anti-PLA2R antibody titer of 61.44 (14.35, 193.24) RU/ml, serum C3 of 1.005 (0.864, 1.150) g/L, and serum C4 of 0.260 (0.214, 0.317) g/L. Kaplan-Meier survival curve showed that the incomplete remission rate of proteinuria in serum C3 > 1.005 g/L group was lower than that in serum C3 ≤ 1.005 g/L group (log-rank χ2=4.757, P=0.029). There was no significant difference in the incomplete remission rate of proteinuria between serum C4 ≤ 0.260 g/L group and serum C4 > 0.260 g/L group (log-rank χ2=3.543, P=0.060). Renal C1q (log-rank χ2=0.167, P=0.683) and C4 (log-rank χ2=1.927, P=0.165) deposition had no significant effects on proteinuria remission in PMN patients. The incomplete remission rate of proteinuria in patients with renal C3 deposition was higher than that in patients without renal C3 deposition (log-rank χ2=7.018, P=0.008). Univariate Cox regression analysis showed that serum C3 level and C3 deposition in renal tissues were influencing factors of incomplete remission of proteinuria (both P<0.05), while adjusting for gender, age, mean arterial pressure, serum anti-PLA2R antibody, serum albumin and 24 h urinary protein, serum C3 ≤ 1.005 g/L ( HR=1.374, 95% CI 1.021-1.849, P=0.036), C3 deposition in renal tissues ( HR=1.949, 95% CI 1.098-3.460, P=0.023), and serum C3 ≤ 1.005 g/L combined with C3 deposition in renal tissues ( HR=1.472, 95% CI 1.093-1.983, P=0.011) were independent influencing factors of incomplete remission of proteinuria. Conclusions:The serum C3 level and C3 deposition in renal tissues are closely related to urinary protein level and proteinuria remission status in PMN patients. The patients with higher urinary protein have higher serum C3. For patients with massive proteinuria, serum C3 ≤ 1.005 g/L, C3 deposition in renal tissues, serum C3 ≤ 1.005 g/L combined with C3 deposition in renal tissues are independent risk factors of incomplete remission of proteinuria.

Objective To evaluate the molluscicidal activity of a novel molluscicide pyriclobenzuron against Oncomelania hupensis robertsoni in the mountain regions of Yunan Province, and test its toxicity to fish, so as to provide scientific evidence for the extensive application of this molluscicide in schistosomiasis-endemic foci of Yunan Province. Methods In the laboratory and snail-breeding field of Dali Prefecture, Yunnan Province, the molluscicidal activity of 5% wettable powder of pyriclobenzuron sulphate (25% PBU) against O. hupensis robertsoni was assessed by using the immersion and spraying method, and the acute toxicity of 25% PBU to carp fries was tested, while 25% wettable powder of niclosamide ethanolamine salt (50% WPNES) served as a control. Results The 1-, 2- and 3-day 25% PBU LC50 and LC90 values were 0.47, 0.25 and 0.23 mg/L, and 1.54, 0.61 and 0.49 mg / L for O. h. robertsoni by using the immersion method in laboratory, and immersion with 25% PBU at 1.0 mg / L for 1 day achieved a comparable molluscicidal efficacy in relative to 50% WPNES at 1.0 mg/L. Spraying with 25% PBU at 4.0 g/m² achieved 1-, 3- and 7-day snail mortalities of 64.23%, 96.67% and 100.00% in laboratory, respectively, which were not significantly different from those caused by treatment with 50% WPNES at 1.0 g/m² (all P values > 0.05). One-day field immersion with 25% PBU at doses of 1, 2 and 4 g/m³ resulted in snail mortalities of 90.00%, 93.33% and 100.00%, respectively, which were not significantly different from those caused by treatment with 50% WPNES at 1.0 g/m³ (all P values > 0.05), and 3-day field spraying with 25% PBU at doses of 2.0 and 4.0 g/m² caused snail mortalities of 86.36% and 87.72%, respectively, which were not significantly different from those caused by 50% WPNES treatment (both P values > 0.05). The 24-, 48- and 72-hour LC50 values of 25% PBU to carp fries were 29.38, 24.62 and 23.38 mg/L, respectively, and no fish death was observed within 72 hours of exposure to 25% PBU at a concentration of 17.5 mg/L and lower. Conclusion 25% PBU is a novel, highly potent and environment-friendly molluscicide that is feasible in fish ponds, and the recommended dose is 1 g/m³ for field immersion and 2 g/m² for field spraying in schistosomiasis-endemic areas of Yunnan Province.