A sensitive determination method for sulfur mustard ( HD ) metabolites thiodiglycol ( TDG ) in rabbit urine was established and validated using isotope dilution negative ion chemical ionization ( NICI) gas chromatography-mass spectrometry ( GC-MS ) , in which deuterated thiodiglycol ( TDG-d8 ) was used as internal standard. Two solid-phase extraction ( SPE) steps were established and optimized in order to reduce the interfering backgrounds, one was used to extract thiodiglycol ( TDG ) from urine with self-assemblied Florisil SPE cartridges, another cleaning treatment of the by-products after pentafluorobenzoyl chloride (PFBZ) derivatization. The results showed that the limits of detection quantitation of this method were 0. 1 and 0. 3 μg/L, respectively. The exposure time-response relationship and exposure dose-response relationship of TDG in rabbit urine were studied after rabbit skin exposure to sulfur mustard (HD, 0. 02-0. 15 LD50). The TDG levels in the rabbit urine increased rapidly during the first day after application and then decreased over time for all dosage groups. A secondary release was also noted for the high-dose group, and the duration of high TDG excretion levels was correlated positively with the HD dosage levels. We thus concluded that abnormally high levels of TDG in urine could be used as a clear diagnostic indicator of HD exposure.

OBJECTIVE To prepare the glutathione adducts of divinylsulfone (DVS), which is an important oxidative metabolism product of SM in vivo, and to investigate their reactive capability with DNA in vitro. METHODS The mustard sulfoxide (SMO) and mustard sulfone (SMO2) were prepared by oxidation reaction using HNO3 and KMnO4 as oxidants, respectively. Then, DVS was prepared through dechlorination reaction using CaCO3 under alkaline conditions. Furthermore, the DVS-GSH adduct and DVS-GSH-purine adducts were prepared and identified using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) and nuclear magnetic resonance (NMR). Finally, the adduct reac?tion process of DVS with GSH was monitored using UPLC-MS/MS. RESULTS The DVS-GSH and GSH-DVS-purine adducts were obtained through preparative HPLC and characterized using NMR and high-resolution MS. In aqueous solution, the reactive activity of DVS with GSH was significantly higher than that of SM, and the DVS-GSH adduct had high or reactive activity, which could produce a series of adducts with adenine and guanine in DNA, and the abundance of the adenine adducts was higher than that of the guanine. CONCLUSION DVS-GSH adducts still have high reactive activity with DNA, and more attention should be paid to its potential damage to DNA.

OBJECTIVE: To investigate the behavioral and ultrastructural changes of intrathecal administration of different concentrations of ropivacaine for 12 h. METHODS: Thirty male SD rats were randomly divided into 5 groups (6 rats in each group):group N (control), group A (ropivacaine 0.25%), group B (ropivacaine 0.5%), group C (ropivacaine 0.75%),and group D (ropivacaine 1.0%). A polyurethane microcatheter was inserted into the lumbar subarachnoid space 8 cm according to Yakshos intrathecal administration. The rats in group N received saline 0.12 mL/kg for 8 times at 1.5 h interval through the catheter, and the rats in the other groups received different concentrations of ropivacaine in the same way as in group N. The poster paw withdrawal latency to heat (PWHL) and mechanical stimulation (von Fray filament) (PWML) were measured the day before the intrathecal administration and 12 hours after the first intrathecal administration of ropivacaine. Motor function (MF) was measured after the last intrathecal administration. After the behavior test, the rats were sacrificed and the lumber segments of the spinal cord were immediately removed for electron microscopic examination. RESULTS: A total hind limb paralysis was seen at 30 seconds and intramuscular strain gradually came back 10~60 minutes after the intrathecal administration of ropivacaine in group A, B, C, and D, but not in group N. The recovery time of motor block of group A was the shortest (P<0.05), that of group D was the longest,and that of group B and C was between group A and D. Intrathecal administration of different concentrations of ropivacaine did not affect the percent maximum possible effect (%MPE) of PWHL and PWML. Electron microscopic examination showed that the spinal cords were normal in group N and A, slight edema of mitochondria and endoplasmic reticulum (ER) in group B, loosened fibrous layers in medullary sheath, edema and local degeneration of neuraxis in group C,and shrinkage of nuclear membrane, serious edema of ER, vacuolus change of mitochondria and local demyelination in group D. CONCLUSION Ropivacaine (0.5%, 0.75%, and 1.0%) administered intrathecally for 12 hours causes different degrees of ultrastructural changes in the spinal cord depending on concentrations.
Amides ; administration & dosage ; toxicity ; Anesthetics, Local ; administration & dosage ; toxicity ; Animals ; Behavior, Animal ; drug effects ; Dose-Response Relationship, Drug ; Injections, Spinal ; Male ; Microscopy, Electron ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Ropivacaine ; Spinal Cord ; ultrastructure

Objective:Applying bibliometrics to analyze the research history, hotspots and trends of Traditional Chinese Medicine (TCM) interventions in edema-related diseases, and to provide reference for the revision of diagnostic and therapeutic criteria for edema in TCM.Methods:The literature about edema treated with TCM was retrieved from CNKI, Wanfang, VIP, CBM from 1 st. Jan 1995 to 25 th. May 2022. CiteSpace and VOSviewer softwares were used to draw the knowledge map, and analyze the co-occurrence relationship and clustering characteristics of the institution, author, keywords and mechanism hotspots. Results:Totally 3 198 articles were included. The annual number of documents issued generally shows a spiral rise trend. Liaoning University of Chinese Medicine published the most articles. Core authors published the most articles were Sun Wei (13 articles); the team with the highest cooperation intensity was Yang Hongtao's team; keywords formed 7 clusters. Hotspot mechanisms included metabolic disorders, immune balance, anti-inflammation, calcium and phosphorus metabolism. Keywords in the past 7 years were membranous nephropathy, chronic heart failure, diabetes, lymphedema etc.Conclusions:The attention paid to the intervention of TCM in the field of edema is generally on the rise, and has decreased in the past two years. The research categories focus on the experience of famous doctors, clinical trials, and mechanistic studies, and nephropathy-related edema has been the focus of research; diabetic nephropathy, chronic heart failure, metabolomics, and immunotherapy are expected to be the focus of attention in the next stage.

Objective : investigate the correlation between the ossification of the styloid hyoid ligament and Bell’s facial paralysis and provide a reference for clinical diagnosis and treatment. Methods: A case of ossification of the bilateral stylohyoid ligament with Bell's facial palsy caused by ossification of the bilateral stylohyoid ligament was diagnosed by clinical manifestations, differential diagnosis and imaging examination. The surgical plan was determined, and combined surgical resection of the ossified area of the styloid hyoid ligament and the greater horn of the hyoid was performed. Postoperative cefoxitin sodium anti-inflammatory treatment, methylprednisolone hormone treatment, acyclovir antiviral treatment, mecobalamin nutritional neurotherapy, and the relevant literature were analyzed. Results: The patient experienced pain when swallowing before surgery, disappearance of right frontal ridges, incomplete eyelid closure, and ptosis of mouth corners. An MRI scan of the brain excluded intracranial space-occupying lesions and resulted in the diagnosis of Bell’s facial paralysis. High-resolution CT of the styloid process confirmed ossification of the styloid hyoid ligament. Styloid process shortening and partial hyoid resection were performed under general anesthesia. Half a month after discharge, the symptoms of sore throat and pain in swallowing disappeared, facial nerve function recovered well, right eyelid closure function recovered well, and right mouth droop improved. The facial nerve function basically returned to normal after 1 month of follow-up. A review of the relevant literature showed that ossification of the stylohyoid ligament to form pseudojoint dilation can locally stimulate the peripheral facial nerve and lead to facial paralysis symptoms. Conclusion Ossification of the styloid hyoid ligament is usually characterized by pharyngeal pain, which can be confirmed by imaging examination. Ossification of the styloid hyoid ligament with facial paralysis is rare in the clinic, so it is necessary to make a clear diagnosis and treat the symptoms.

Objective:To analyze the efficacy evaluation criteria of the existing TCM treatment for edema RCT research, and to provide reference for the construction of unified standards.Methods:The batabases CNKI, WanFang Data, VIP, CBM, Pubmed and Web of Science were retrieved. The randomized controlled trials for the treatment of edema of TCM research, from September 1, 1993 to July 31, 2022, were screened and included. The content of efficacy evaluation, performed statistics on evaluation standard, the curative effect evaluation indexes, as well as standard composition, usage were extracted. We analyzed the characteristics, application and problems of the existing efficacy evaluation criterion.Results:A total of 123 Chinese articles were included. The included literature involved nephrogenic edema, cardiogenic edema, idiopathic edema, apoplexy limb edema and other edema. In recent years, randomized controlled trials on the treatment of edema by TCM have mainly used four efficacy evaluation criteria. Of which the Guidelines for Clinical Research on New Chinese Medicines (Trial) in 2002 had the highest utilization rate of 29.27%. Secondly, the utilization rate of Standard for Diagnosis and Curative Effect of TCM Diseases and Syndromes was 21.14%. The rest of the criteria were used by less than 6%. While 39.02% of the literature did not use the standards or used self-designed standards. Among the composition of efficacy evaluation indices, the application rate of TCM syndrome or symptom efficacy index was the highest (91.87%), the utilization rate of the Minnesota Living with Heart Failure Questionnaire Indicators was only 4.88%; biochemical tests accounted for a large proportion of Western medical indicators, while the measurement of edema severity was rarely applied. Conclusions:At present, the evaluation criteria of edema curative effect are diversified and insufficiently popularized, which need to be further screened and improved. It is suggested to construct a TCM edema efficacy evaluation model based on the characteristics of edema syndrome, comprehensively evaluate the efficacy from multiple dimensions such as TCM syndromes, western medicine indicators, and quality of life, and improve the scientific indicators.

Objective To investigate the effect of Huatan Qushi Huoxue prescription on lipopolysaccharide (LPS)-induced pyroptosis of RAW264.7 cells and its mechanism. Methods An in vitro cell model of LPS-induced activated RAW264.7 was established and divided into blank group, model group, high-, middle-, and low-dose Huatan Qushi Huoxue prescription groups, and control group. The corresponding drug-containing serum intervention was performed for 24 hours. A scanning electron microscope was used to observe cell morphology, and immunofluorescence assay was used to perform quantitative localization of GSDMD-N. Results The cells in the blank group were round and regular in shape with smooth surface, and those in the control group were swollen, with folds on the surface and gaps in the capsule, which were consistent with the morphology of cell pyroptosis. The cells in the control group had bubbles on the surface with obvious pseudopodia and pores in cell membrane, and those in the high-dose group were not swollen and had a rough surface with pseudopodia, with no obvious pores in cell membrane. The cells in the low- and middle-dose groups were swollen and had a rough surface of cell membrane with pores and pseudopodia. Immunofluorescence assay showed that compared with the blank group, the model group had a significant increase in the positive staining intensity of GSDMD-N, and compared with the model group, the control group and the Traditional Chinese medicine group had a reduction in the positive staining intensity of GSDMD-N. Conclusion Huatan Qushi Huoxue prescription can improve the pyroptosis of macrophages and reduce the expression of GSDMD-N.

Objective To investigate the effect of Huatan Qushi Huoxue prescription on the ultrastructure of hepatocyte mitochondria in a rat model of nonalcoholic steatohepatitis (NASH). Methods A total of 48 male Sprague-Dawley rats were randomly divided into blank group, model group, Yishanfu group, and Huatan Qushi Huoxue prescription group, with 12 rats in each group. The rats in the model group and the drug groups were administered and modeled since week 2; the rats in the blank group were given normal diet, and those in the other three groups were given high-fat diet. Based on dose conversion between human and animal, the equivalent dose of Huatan Qushi Huoxue prescription was 1.26 g/100 g body weight, and the equivalent dose of polyene phosphatidylcholine capsules (Yishanfu) was 0.014 18 g/100 g body weight. The rats in the model group were given 0.9% sodium chloride by gavage, those in the Yishanfu group were given polyene phosphatidylcholine suspension by gavage, and those in the traditional Chinese medicine group were given the granules of Huatan Qushi Huoxue prescription by gavage, once a day for 10 consecutive weeks. A transmission electron microscope was used to observe liver ultrastructure and perform a quantitative analysis. A one-way analysis of variance was used for comparison of continuous data between multiple groups; for further pairwise comparison, the least significant difference t -test was used for data with homogeneity of variance, and the Dunnett's T3 was used for data with heterogeneity of variance. Results The model group had a large number of lipid droplets accumulated in hepatocytes, changes in mitochondrial morphology and structure, and reductions in the number of mitochondria and endoplasmic reticulum. The Huatan Qushi Huoxue prescription group had a significant reduction in lipid droplets in hepatocytes and significant increases in the number of mitochondria and endoplasmic reticulum compared with the model group, with intact mitochondrial membrane and structure. The Yishanfu group had a reduction in lipid droplets in hepatocytes, an increase in the number of mitochondria, and a reduction in the number of endoplasmic reticulum, with relatively intact mitochondrial membrane and structure. The quantitative analysis showed that compared with the blank group, the model group had a significant increase in the area of lipid droplets and a significant reduction in mitochondria, with a significant difference in mitochondrial density between the two groups (all P < 0.01); after drug intervention, the Yishanfu group had a significant reduction in the area of lipid droplets and a significant increase in the number of mitochondria, with a significant difference in mitochondrial density between the Yishanfu group and the model group (all P < 0.01); compared with the Yishanfu group, the traditional Chinese medicine group had a significantly greater reduction in the area of lipid droplets and a significant increase in the number of mitochondria, with a significant difference in mitochondrial density between the two groups (all P < 0.05). Conclusion Huatan Qushi Huoxue prescription can improve lipid accumulation, increase mitochondrial density, and protect mitochondrial structure and function, with a better clinical effect than Yishanfu.

Pancreatogenic diabetes is a type of diabetes mellitus secondary to exocrine pancreatic disease， and it was officially proposed by the American Diabetes Association in 2014， with chronic pancreatitis as the most common etiology， followed by pancreatic cancer. At present， the misdiagnosis rate of this disease is extremely high， and patients with pancreatogenic diabetes have a higher risk of death and readmission than patients with type 2 diabetes. Therefore， it is of great significance to fully understand， correctly identify， and diagnose pancreatogenic diabetes in the early state， so as to reduce the disability rate and mortality rate of this disease. This article reviews the advances in the possible pathogenesis， diagnosis， treatment， and management of pancreatic diabetes secondary to pancreatitis and pancreatic cancer.

Ferroptosis is a type of iron-dependent cell death driven by lipid peroxidation, and its mechanism is associated with iron homeostasis imbalance, lipid peroxidation, and slC7A11-GSH-GPX4 antioxidant system. Ferroptosis plays a key role in the development and progression of nonalcoholic fatty liver disease (NAFLD)/nonalcoholic steatohepatitis (NASH), and inhibition of ferroptosis can almost completely inhibit the development of NASH. This article reviews the research advances in the mechanism of ferroptosis and its role in NAFLD/NASH and proposes the research strategies and technical means for ferroptosis, so as to provide a reference for research on the mechanism of NAFLD/NASH.