1.Mechanism of Huanglian Jiedu Decoction in treatment of type 2 diabetes mellitus based on intestinal flora.
Xue HAN ; Qiu-Mei TANG ; Wei WANG ; Guang-Yong YANG ; Wei-Yi TIAN ; Wen-Jia WANG ; Ping WANG ; Xiao-Hua TU ; Guang-Zhi HE
China Journal of Chinese Materia Medica 2025;50(1):197-208
The effect of Huanglian Jiedu Decoction on the intestinal flora of type 2 diabetes mellitus(T2DM) was investigated using 16S rRNA sequencing technology. Sixty rats were randomly divided into a normal group(10 rats) and a modeling group(50 rats). After one week of adaptive feeding, a high-fat diet + streptozotocin was given for modeling, and fasting blood glucose >16.7 mmol·L~(-1) was considered a sign of successful modeling. The modeling group was randomly divided into the model group, high-, medium-, and low-dose groups of Huanglian Jiedu Decoction, and metformin group. After seven days of intragastric treatment, the feces, colon, and pancreatic tissue of each group of rats were collected, and the pathological changes of the colon and pancreatic tissue of each group were observed by hematoxylin-eosin staining. The changes in the intestinal flora structure of each group were observed by the 16S rRNA sequencing method. The results showed that compared with the model group, the high-, medium-, and low-dose of Huanglian Jiedu Decoction reduced fasting blood glucose levels to different degrees and showed no significant changes in body weight. The number of islet cells increased, and intestinal mucosal damage attenuated. Alpha diversity analysis revealed that Huanglian Jiedu Decoction reduced the abundance and diversity of intestinal flora in rats with T2DM; at the phylum level, low-and mediam-dose of Huanglian Jiedu Decoction reduced the abundance of Bacteroidota, Proteobacteria, and Desulfobacterota and increased the abundance of Firmicute and Bacteroidota/Firmicutes, while the high-dose of Huanglian Jiedu Decoction increased the relative abundance of Proteobacteria and Bacteroidota/Firmicutes ratio, and decreaseal the relative; abundance of Firmicute; at the genus level, Huanglian Jiedu Decoction increased the relative abundance of Allobaculum, Blautia, and Lactobacillus; LEfse analysis revealed that the biomarker of low-and medium-dose groups of Huanglian Jiedu Decoction was Lactobacillus, and the structure of the intestinal flora of the low-dose group of Huanglian Jiedu Decoction was highly similar to that of the metformin group. PICRUSt2 function prediction revealed that Huanglian Jiedu Decoction mainly affected carbohydrate and amino acid metabolic pathways. It suggested that Huanglian Jiedu Decoction could reduce fasting blood glucose and increase the number of islet cells in rats with T2DM, and its mechanism of action may be related to increasing the abundance of short-chain fatty acid-producing strains and Lactobacillus and affecting carbohydrate and amino acid metabolic pathways.
Animals
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Drugs, Chinese Herbal/administration & dosage*
;
Diabetes Mellitus, Type 2/metabolism*
;
Gastrointestinal Microbiome/drug effects*
;
Rats
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Male
;
Rats, Sprague-Dawley
;
Humans
;
Bacteria/drug effects*
;
Blood Glucose/metabolism*
2.Effect and mechanism of alkaloids from Portulacae Herba on ulcerative colitis in mice based on TLR4/MyD88/NF-κB signaling pathway.
Jia-Hui ZHENG ; Ying-Ying SONG ; Tian-Ci ZHANG ; Wen-Ting WANG ; Zhi-Ping YANG ; Jin-Xia AI
China Journal of Chinese Materia Medica 2025;50(4):874-881
This study investigated the functions and regulatory mechanism of Portulacae Herba and its chemical components on the Toll-like receptor 4(TLR4)/myeloid differentiation primary response 88(MyD88)/nuclear factor kappa B(NF-κB) inflammatory signaling pathway in the colon tissue of mice with dextran sodium sulfate(DSS)-induced ulcerative colitis(UC). A total of 35 mice were randomly divided into groups, including a blank group, a model group, a mesalazine group(0. 5 g·kg~(-1)), and low, medium,and high dose alkaloids from Portulacae Herba groups(9, 18, 36 mg·kg~(-1)), and a combination treatment group, with 5 mice in each group. The blank group was given purified water, while the other groups were continuously given a 3% DSS solution for 7 days to induce the UC model. From day 8 onwards, the treatment group received oral gavage according to the prescribed doses for 14 days. The overall condition, body weight, stool characteristics, and presence of blood in the stool were recorded daily. After the experiment, the disease activity index(DAI) was assessed for each group, and colon length was measured. Histopathological changes in colon tissue were examined using hematoxylin-eosin(HE) staining. The levels of pro-inflammatory cytokines, tumor necrosis factor-α(TNF-α),and interleukin-1β( IL-1β) in serum were measured by enzyme-linked immunosorbent assay( ELISA). The protein and m RNA expression of TLR4, MyD88, and NF-κB in colon tissue were measured using Western blot and quantitative real-time PCR(qPCR).Compared to the blank group, the model group showed a significant decrease in body weight, a notable increase in DAI scores, a significant shortening of colon length, and evident histopathological damage. The levels of inflammatory cytokines TNF-α and IL-1β in the serum were significantly elevated, and the protein and m RNA expression of TLR4, MyD88, and NF-κB in colon tissue were significantly up-regulated. In contrast, the alkaloids from Portulacae Herba treatment groups significantly improved symptoms and reduced body weight loss in mice, decreased DAI scores, alleviated colon shortening, lowered serum levels of TNF-α and IL-1β,significantly down-regulated the expression levels of TLR4, MyD88, and NF-κB proteins and genes in colon tissue, as well as reduced histopathological damage. Therefore, the study suggests that alkaloids from Portulacae Herba can alleviate intestinal inflammation damage in DSS-induced UC mice, with its mechanism involving the TLR4/MyD88/NF-κB signaling pathway.
Animals
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Colitis, Ulcerative/immunology*
;
Toll-Like Receptor 4/immunology*
;
Myeloid Differentiation Factor 88/metabolism*
;
Mice
;
NF-kappa B/metabolism*
;
Signal Transduction/drug effects*
;
Male
;
Alkaloids/administration & dosage*
;
Drugs, Chinese Herbal/administration & dosage*
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Humans
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Female
;
Colon/metabolism*
;
Disease Models, Animal
3.Huotan Jiedu Tongluo Decoction inhibits ferroptosis by regulating Nrf2/GPX4 pathway to ameliorate atherosclerotic lesions in ApoE~(-/-) mice.
Di GAO ; Teng-Hui TIAN ; Ke-Ying YU ; Xiao SHAO ; Wen XUE ; Zhi-Xuan ZHAO ; Yue DENG
China Journal of Chinese Materia Medica 2025;50(7):1908-1919
The purpose of this study was to clarify the effect of Huotan Jiedu Tongluo Decoction on atherosclerosis(AS) injury in ApoE~(-/-) mice by regulating the ferroptosis pathway. Seventy-five ApoE~(-/-) mice were randomly divided into model group, low-, medium-, and high-dose of Huotan Jiedu Tongluo Decoction groups, and evolocumab group(n=15), and 15 C57BL/6J mice were selected as the blank group. Mice in the blank group were fed with a normal diet, and those in the other groups were fed with a high-fat diet to induce AS. From the 9th week, mice in Huotan Jiedu Tongluo Decoction groups were administrated with Huotan Jiedu Tongluo Decoction at corresponding doses by gavage, and those in the blank group and the model group were given an equal volume of distilled water. Mice in the evolocumab group were treated with evolocumab 18.2 mg·kg~(-1 )by subcutaneous injection every 2 weeks. After 8 weeks of continuous intervention, oil red O staining and hematoxylin-eosin(HE) staining were employed to observe the lipid deposition and plaque formation in the aortic root. Masson staining was used to evaluate the collagen content in the aortic root. The serum levels of total cholesterol(TC), triglycerides(TG), high-density lipoprotein cholesterol(HDL-C), and low-density lipoprotein cholesterol(LDL-C) were determined by biochemical kits. The levels of Fe~(2+), superoxide dismutase(SOD), malondialdehyde(MDA), and glutathione(GSH) in the aorta were measured by colorimetry. The protein and mRNA levels of nuclear factor erythroid 2-related factor 2(Nrf2), glutathione peroxidase 4(GPX4), solute carrier family 7 member 11(SLC7A11), and acyl-CoA synthetase long chain family member 4(ACSL4) in the aorta were detected by Western blot and RT-qPCR, respectively. The expression of Nrf2, GPX4, and SLC7A11 was localized by immunofluorescence. The results showed that low-, medium-, and high-dose Huotan Jiedu Tongluo Decoction reduced the plaque formation of aortic root and increased the collagen content in AS mice. At the same time, Huotan Jiedu Tongluo Decoction improved the lipid metabolism by lowering the levels of TC, LDL-C, and TG and elevating the level of HDL-C in the serum. Huotan Jiedu Tongluo Decoction enhanced the antioxidant capacity by elevating the levels of GSH and SOD and lowering the level of MDA in the aorta and inhibiting the accumulation of Fe~(2+) in the aorta. In addition, Huotan Jiedu Tongluo Decoction up-regulated the protein and mRNA levels of Nrf2, GPX4, and SLC7A11, while down-regulating the protein and mRNA levels of ACSL4. In summary, Huotan Jiedu Tongluo Decoction can effectively alleviate AS lesions in ApoE~(-/-) mice by activating the Nrf2/GPX4 pathway, reducing lipid peroxidation, and inhibiting ferroptosis.
Animals
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Ferroptosis/drug effects*
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Atherosclerosis/metabolism*
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Drugs, Chinese Herbal/administration & dosage*
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NF-E2-Related Factor 2/genetics*
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Mice
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Mice, Inbred C57BL
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Apolipoproteins E/metabolism*
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Male
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Phospholipid Hydroperoxide Glutathione Peroxidase/genetics*
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Signal Transduction/drug effects*
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Humans
;
Mice, Knockout
4.Application of Assessment Scales in Palliative Care for Glioma: A Systematic Review.
Zhi-Yuan XIAO ; Tian-Rui YANG ; Ya-Ning CAO ; Wen-Lin CHEN ; Jun-Lin LI ; Ting-Yu LIANG ; Ya-Ning WANG ; Yue-Kun WANG ; Xiao-Peng GUO ; Yi ZHANG ; Yu WANG ; Xiao-Hong NING ; Wen-Bin MA
Chinese Medical Sciences Journal 2025;40(3):211-218
BACKGROUND AND OBJECTIVE: Patients with glioma experience a high symptom burden and have diverse palliative care needs. However, the assessment scales used in palliative care remain non-standardized and highly heterogeneous. To evaluate the application patterns of the current scales used in palliative care for glioma, we aim to identify gaps and assess the need for disease-specific scales in glioma palliative care. METHODS: We conducted a systematic search of five databases including PubMed, Web of Science, Medline, EMBASE, and CINAHL for quantitative studies that reported scale-based assessments in glioma palliative care. We extracted data on scale characteristics, domains, frequency, and psychometric properties. Quality assessments were performed using the Cochrane ROB 2.0 and ROBINS-I tools. RESULTS: Of the 3,405 records initially identified, 72 studies were included. These studies contained 75 distinct scales that were used 193 times. Mood (21.7%), quality of life (24.4%), and supportive care needs (5.2%) assessments were the most frequently assessed items, exceeding half of all scale applications. Among the various assessment dimensions, the Distress Thermometer (DT) was the most frequently used tool for assessing mood, while the Short Form-36 Health Survey Questionnaire (SF-36) was the most frequently used tool for assessing quality of life. The Mini Mental Status Examination (MMSE) was the most common tool for cognitive assessment. Performance status (5.2%) and social support (6.8%) were underrepresented. Only three brain tumor-specific scales were identified. Caregiver-focused scales were limited and predominantly burden-oriented. CONCLUSIONS: There are significant heterogeneity, domain imbalances, and validation gaps in the current use of assessment scales for patients with glioma receiving palliative care. The scale selected for use should be comprehensive and user-friendly.
Humans
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Glioma/psychology*
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Palliative Care/methods*
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Quality of Life
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Psychometrics
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Brain Neoplasms/psychology*
5.Tibial transverse transport combined with antibiotic-loaded bone cement for the treatment of diabetic foot ulcer.
Wen CHEN ; Li TIAN ; Tao LIU ; Zhi-Jun HE ; Yan LI ; Jin-Peng LI
China Journal of Orthopaedics and Traumatology 2025;38(9):878-883
OBJECTIVE:
To compare clinical efficacy of tibial transverse transport(TTT) combined with antibiotic-loaded bone cement (ABC) and TTT in treating diabetic foot ulcer (DFU).
METHODS:
A retrospective analysis was conducted on 60 patients with DFU treated from January 2019 to January 2023. They were divided into bone cement group and bone transfer group according to different treatment methods, with 30 patients in each group. There were 20 males and 10 females in bone cement group, aged from 61 to 76 years old with an average of (68.15±4.85) years old;the course of ulcer disease ranged from 7 to 28 months with an average of (15.28±5.52) months;16 patients were grade 3 and 14 patients were grade 4 according to Wagner classification; TTT combined with ABC treatment was performed. There were 22 males and 8 females in bone transfer group, aged from 60 to 75 years old with average of (67.85±4.62) years old;the course of ulcer disease ranged from 6 to 29 months with an average of (14.35±5.21) months;17 patients were grade 3 and 13 patients were grade 4 according to Wagner classification;TTT was performed. The control time of wound infection, duration of antibiotic use, frequency of debridement, weight-bearing time of the affected limb, healing time of ulcer surface and recurrence of infection were compared between two groups. Visual analogue scale (VAS) and ankle brachial index (ABI) between two groups was compared before operation and 2 and 6 months after operation.
RESULTS:
Sixty patients were followed up for 12 to 24 months with average of (17.24±4.42) months. The control time of wound infection, duration of antibiotic use, frequency of debridement, weight-bearing time of the affected limb, and healing time of ulcer surface in bone cement group were (11.02±2.14) days, (12.7±3.5) days, (1.2±0.4) times, (90.02±2.75) days, and (2.32±3.45) months, respectively;while in bone transfer groups were (20.14±3.15) days, (20.4±4.5) days, (2.2±0.8) times, (106.64±8.35) days, and (4.53±3.12) months respectively; bone cement group was superior to bone transfer group, and the differences were statistically significant(P<0.05). Comparisons of VAS and ABI before and after treatment between two groups showed preoperative VAS and ABI in bone cement group were (6.71±0.73) points and (0.25±0.04) respectively, and those in bone transfer group were (6.87±0.17) points and (0.27±0.03) respectively. At 2 months after operation, VAS and ABI in bone cement group were (3.71±0.47) points and (0.61±0.03) respectively, and those in bone transfer group were (3.79±0.70) points and (0.59±0.05) respectively;postoperative VAS and ABI at 6 months in bone cement group were (2.26±0.13) points and (0.80±0.05) respectively, and those in bone transfer group were (2.57±0.17) points and (0.79±0.04) respectively;postoperative VAS and ABI between groups were improved at each time points compared with those of before operation (P<0.05). In bone cement group, there were 2 patients with ulcer recurrence and 1 patient with gangrene;while in bone transfer group, 5 patients with recurrence of infection, 2 patients with recurrence of ulcer and 1 patient with gangrene;the recurrence rate of infection in bone cement group were lower than that in bone transfer group (P<0.05).
CONCLUSION
The combination of TTT and ABC in treating DFU has a good therapeutic effect, which could be shorten the infection control time, ulcer healing time and antibiotic use time, effectively relieve pain, reduce the recurrence rate of infection and improve the quality of life of patients.
Humans
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Male
;
Female
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Aged
;
Bone Cements/therapeutic use*
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Middle Aged
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Anti-Bacterial Agents/therapeutic use*
;
Diabetic Foot/therapy*
;
Retrospective Studies
;
Tibia/surgery*
6."A diamond-shaped" penoplasty technique with or without concurrent suprapubic liposuction for adult-acquired buried penis: clinical outcomes and patient satisfaction rates.
Jing WANG ; Jian NI ; Yang XU ; Wen YU ; Zhi-Peng XU ; Yu-Tian DAI ; Yi-Qiong YANG ; Xiao-Zhi ZHAO
Asian Journal of Andrology 2025;27(1):72-75
Various techniques have been described for reconstructing the skin of the penile shaft; however, no universally accepted standard exists for correcting buried penis in adults. We aimed to describe a new technique for correcting an adult-acquired buried penis through a diamond-shaped incision at the penopubic junction. We retrospectively analyzed data from patients treated with our technique between March 2019 and June 2023 in the Department of Andrology, Nanjing Drum Tower Hospital (Nanjing, China). Forty-two adult males with buried penises, with a mean (±standard deviation [s.d.]) age of 26.6 (±6.6) years, underwent surgery. All patients were obese, with an average (±s.d.) body mass index of 35.56 (±3.23) kg m -2 . In addition to phalloplasty, 32 patients concurrently underwent circumcision, and 28 underwent suprapubic liposuction. The mean (±s.d.) duration of the operation was 98.02 (±13.28) min. The mean (±s.d.) duration of follow-up was 6.71 (±3.43) months. The length in the flaccid unstretched state postoperatively was significantly greater than that preoperatively (mean ± s.d: 5.55±1.19 cm vs 1.94±0.59 cm, P < 0.01). Only minor complications, such as wound disruption (7.1%) and infection (4.8%), were observed. The mean (±s.d.) score of patient satisfaction was 4.02 (±0.84) on a scale of 5. This technique provides excellent cosmetic and functional outcomes with a minimal risk of complications. However, additional clinical studies are needed to evaluate the long-term effects of this procedure.
Humans
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Male
;
Patient Satisfaction
;
Adult
;
Lipectomy/methods*
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Penis/surgery*
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Retrospective Studies
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Treatment Outcome
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Plastic Surgery Procedures/methods*
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Young Adult
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Penile Diseases/surgery*
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Urologic Surgical Procedures, Male/methods*
7.Clinical and Laboratory Characteristics of Streptococcus mitis Causing Bloodstream Infection in Children with Hematological Disease.
Yu-Long FAN ; Guo-Qing ZHU ; Zhi-Ying TIAN ; Yan-Xia LYU ; Zhao WANG ; Ye GUO ; Wen-Yu YANG ; Qing-Song LIN ; Xiao-Juan CHEN
Journal of Experimental Hematology 2025;33(1):286-291
OBJECTIVE:
To investigate the risk factors, clinical characteristics, and bacterial resistance of bloodstream infections caused by Streptococcus mitis in children with hematological disease, so as to provide a reference for infection control.
METHODS:
The clinical information and laboratory findings of pediatric patients complicated with blood cultures positive for Streptococcus mitis from January 2018 to December 2020 in the Institute of Hematology & Blood Diseases Hospital were searched and collected. The clinical characteristics, susceptibility factors, and antibiotic resistance of the children were retrospectively analyzed.
RESULTS:
Data analysis from 2018 to 2020 showed that the proportion of Streptococcus mitis isolated from bloodstream infections in children (≤14 years old) with hematological diseases was the highest (19.91%) and significantly higher than other bacteria, accounting for 38.64% of Gram-positive cocci, and presented as an increasing trend year by year. A total of 427 children tested positive blood cultures, including 85 children with bloodstream infections caused by Streptococcus mitis who tested after fever. Most children experienced a recurrent high fever in the early and middle stages (≤6 d) of neutropenia and persistent fever for more than 3 days. After adjusting the antibiotics according to the preliminary drug susceptibility results, the body temperature of most children (63.5%) returned to normal within 4 days. The 85 children were mainly diagnosed with acute myeloid leukemia (AML), accounting for 84.7%. The proportion of children in the neutropenia stage was 97.7%. The incidence of oral mucosal damage, lung infection, and gastrointestinal injury symptoms was 40%, 31.8%, and 27.1%, respectively. The ratio of elevated C-reactive protein (CRP) and procalcitonin was 65.9% and 9.4%, respectively. All isolated strains of Streptococcus mitis were not resistant to vancomycin and linezolid, and the resistance rate to penicillin, cefotaxime, levofloxacin, and quinupristin-dalfopristin was 10.6%, 8.2%, 9.4%, and 14.1%, respectively. None of children died due to bloodstream infection caused by Streptococcus mitis.
CONCLUSION
The infection rate of Streptococcus mitis is increasing year by year in children with hematological diseases, especially in children with AML. Among them, neutropenia and oral mucosal damage after chemotherapy are high-risk infection factors. The common clinical symptoms include persistent high fever, oral mucosal damage, and elevated CRP. Penicillin and cephalosporins have good sensitivity. Linezolid, as a highly sensitive antibiotic, can effectively control infection and shorten the course of disease.
Humans
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Child
;
Streptococcal Infections/microbiology*
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Retrospective Studies
;
Hematologic Diseases/complications*
;
Streptococcus mitis
;
Drug Resistance, Bacterial
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Risk Factors
;
Microbial Sensitivity Tests
;
Anti-Bacterial Agents
;
Female
;
Male
;
Bacteremia/microbiology*
;
Child, Preschool
;
Adolescent
8.Diagnosis and clinical characteristics of atypical severe pneumonia caused by Chlamydia abortus
Hai-Wen ZENG ; Qiao-Li CHEN ; Zhi-Rong DING ; Wei-Wen CHEN ; Tian-Lai LIN
Chinese Journal of Infection Control 2024;23(3):358-363
Objective To explore the diagnosis and clinical characteristics of atypical severe pneumonia caused by Chlamydia abortus(C.abortus).Methods Clinical data of 4 patients diagnosed with atypical severe pneumonia caused by C.abortus in a hospital from January 2021 to November 2022 were collected.Clinical characteristics,dia-gnosis and treatment,and precautions of the disease were comprehensively analyzed.Results All 4 patients were male,aged 63-73 years old,with acute onset,high fever,cough and expectoration.Three patients had a history of contact with poultry,one patient had a history of contact with abortion goat.The interval between the emerging of clinical symptoms and the onset of acute respiratory failure in 4 patients was 1-6 days,and the oxygenation index(PaO2/FiO2)at admission was less than 200 mmHg,which gradually decreased with the progression of the disease,active support with a ventilator was necessary.Two patients had an increase in white blood cell count,4 had an in-crease in neutrophil percentage,3 had a mild decrease in platelet count.Among 4 patients,2,2,3 and 4 patients showed elevated levels of aspartate aminotransferase,alanine aminotransferase,creatine kinase,and serum creati-nine respectively,2 patients had mild hyponatremia,4 patients showed significant increase in C-reactive protein,procalcitonin,and interleukin-6 levels.Four patients'chest CT findings showed main involvement of single or mul-tiple lung lobes,with exudation and consolidation,and later involvement of multiple lobes of lung.The metageno-mic next-generation sequencing of bronchoalveolar lavage fluid detected the DNA sequence of C.abortus.Based on the clinical manifestations,contact history,chest CT,and metagenomic next-generation sequencing results of 4 pa-tients,the diagnosis was C.abortus.atypical severe pneumonia.After timely adjustment of the treatment of anti-in-fection regimen based on doxycycline,the patients'condition improved and were discharged.Conclusion C.abor-tus may also cause human pneumonia,which can lead to serious clinical outcome after infection.Patient had a histo-ry of animal contact should be alert to such diseases.Metagenomic next-generation sequencing can detect C.abortus.
9.Toxicokinetics of MDMA and Its Metabolite MDA in Rats
Wei-Guang YU ; Qiang HE ; Zheng-Di WANG ; Cheng-Jun TIAN ; Jin-Kai WANG ; Qian ZHENG ; Fei REN ; Chao ZHANG ; You-Mei WANG ; Peng XU ; Zhi-Wen WEI ; Ke-Ming YUN
Journal of Forensic Medicine 2024;40(1):37-42
Objective To investigate the toxicokinetic differences of 3,4-methylenedioxy-N-methylamphetamine(MDMA)and its metabolite 4,5-methylene dioxy amphetamine(MDA)in rats af-ter single and continuous administration of MDMA,providing reference data for the forensic identifica-tion of MDMA.Methods A total of 24 rats in the single administration group were randomly divided into 5,10 and 20 mg/kg experimental groups and the control group,with 6 rats in each group.The ex-perimental group was given intraperitoneal injection of MDMA,and the control group was given intraperi-toneal injection of the same volume of normal saline as the experimental group.The amount of 0.5 mL blood was collected from the medial canthus 5 min,30 min,1 h,1.5 h,2 h,4 h,6 h,8 h,10 h,12 h after administration.In the continuous administration group,24 rats were randomly divided into the experi-mental group(18 rats)and the control group(6 rats).The experimental group was given MDMA 7 d by continuous intraperitoneal injection in increments of 5,7,9,11,13,15,17 mg/kg per day,respectively,while the control group was given the same volume of normal saline as the experimental group by in-traperitoneal injection.On the eighth day,the experimental rats were randomly divided into 5,10 and 20 mg/kg dose groups,with 6 rats in each group.MDMA was injected intraperitoneally,and the con-trol group was injected intraperitoneally with the same volume of normal saline as the experimental group.On the eighth day,0.5 mL of blood was taken from the medial canthus 5 min,30 min,1 h,1.5 h,2 h,4 h,6 h,8 h,10 h,12 h after administration.Liquid chromatography-triple quadrupole tandem mass spectrometry was used to detect MDMA and MDA levels,and statistical software was employed for data analysis.Results In the single-administration group,peak concentrations of MDMA and MDA were reached at 5 min and 1 h after administration,respectively,with the largest detection time limit of 12 h.In the continuous administration group,peak concentrations were reached at 30 min and 1.5 h af-ter administration,respectively,with the largest detection time limit of 10 h.Nonlinear fitting equations for the concentration ratio of MDMA and MDA in plasma and administration time in the single-administration group and continuous administration group were as follows:T=10.362C-1.183,R2=0.974 6;T=7.397 3C-0.694,R2=0.961 5(T:injection time;C:concentration ratio of MDMA to MDA in plasma).Conclusions The toxicokinetic data of MDMA and its metabolite MDA in rats,obtained through single and continuous administration,including peak concentration,peak time,detection time limit,and the relationship between concentration ratio and administration time,provide a theoretical and data foundation for relevant forensic identification.
10.Glutathione Detection Method Based on Electron Paramagnetic Resonance Spectroscopy
Zhi-Wen WANG ; Jian KUANG ; Ao-Kun LIU ; Ruo-Tong WEI ; Lu YU ; Chang-Lin TIAN
Progress in Biochemistry and Biophysics 2024;51(11):3034-3045
ObjectiveGlutathione (γ-glutamyl-L-cysteinylglycine, GSH) is the most abundant non-protein compound containing sulfhydryl (―SH) groups in cells. It serves as a source of reducing equivalents, effectively neutralizing harmful reactive substances, and playing a crucial role in maintaining cellular redox balance. Therefore, sensitive detection and accurate measurement of GSH levels in tissues are of great importance. In this work, we presents a novel method for GSH detection utilizing electron paramagnetic resonance (EPR) spectroscopy. MethodsInitially, ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate acid)) solution was mixed with K2S2O8 solution and reacted in the dark for 12 to 16 h to prepare ABTS·+ solution, which was then quantified using UV-Vis spectroscopy. Subsequently, the concentration of glutathione (GSH) was determined based on the changes in the EPR signal of ABTS·+. On this basis, the optimal reaction time and temperature were explored to establish a standard equation correlating the EPR signal intensity of ABTS·+ with GSH concentration. Finally, the derived standard curve was employed to quantitatively analyze the GSH concentration in whole blood from C57BL/6J mice, and the results were compared with those reported in the literature to verify the accuracy of the method. ResultsThe experimental results demonstrate that this method has a linear detection range from50 nmol/L to 15 μmol/L for GSH, spanning two orders of magnitude, with a limit of detection (LOD) at0.50 nmol/L. The measured GSH content in mouse whole blood is (10 660±706) nmol/g Hb, which agrees with the value of (11 200±237) nmol/g Hb as previously reported. Furthermore, a similar method was developed for detection of glutathione disulfide (GSSG) at higher reaction temperature. ConclusionThis article presents a novel assay for the rapid detection of GSH using the intensity of EPR signal from ABTS·+ as indicator. This method demonstrates enhanced detection sensitivity and a broader linear range compared to conventional colorimetric methods. Furthermore, we have extended the application of this method to detect GSH content in blood samples efficiently and accurately, offering valuable information for assessing tissue redox balance, thus holding significant potentials.

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