1.Whole genome sequence characteristics of 13 dengue virus serotype 2 strains isolated from Guangzhou in 2023
MA Jinzhe ; LIANG Yufeng ; ZHOU Zhijian
China Tropical Medicine 2025;25(1):110-
Objective To analyze the whole genome sequence characteristics of dengue serotype 2 virus (DENV-2) strains isolated from Guangzhou in 2023, providing new foundational data to support local dengue fever prevention and control efforts. Methods Dengue viruses were isolated from serum samples of dengue fever patients using C6/36 cells. Whole genome sequencing of dengue virus was performed using Nanopore sequencing technology (Nanopore platform). The sequencing data were assembled utilizing IPH-NANO v1.0 software, and homology analysis, phylogenetic analysis, and amino acid site variation analysis of the dengue virus genome sequence were conducted using software such as BioEdit7.0.9.0, MEGA11, and iqTree1.6.12. Results A total of 13 DENV-2 isolates were obtained from the serum samples of dengue fever patients in Guangzhou in 2023, all of which were derived from locally infected cases. The whole genome sequences obtained by sequencing and splicing ranged from 10 429-10 439 nt in length. The nucleotide (amino acid) homology among the 13 isolates was 99.7%-99.9% (99.6%-9.9%). A genome-wide phylogenetic analysis showed that the genotypes of all 13 Guangzhou isolates were identified as the Cosmopolitan genotype and were grouped in the same evolutionary clade as isolates from Réunion (French overseas territory), Djibouti, Kenya, and other regions. Compared with the reference sequence (NC001474), a total of 26 amino acid site variations were identified in the C/prM/E protein regions of the 13 isolates, including 4 variations in the C protein region, 8 in the prM protein region, and 14 in the E protein region. Conclusion The DENV-2 strains isolated from Guangzhou in 2023 exhibited high homology, with closely related isolates primarily originating from countries or regions such as Réunion, Djibouti, and Kenya, it highlights the need for further assessment of the risk of dengue fever importation from East Africa, South Asia, and other regions.
2.Reflections on quarantine and supervision policies of imported experimental animals in Japan
Ning DING ; Jiaqi LIANG ; Hongkun FU ; Ying WANG ; Chunxia WANG ; Jiao REN ; Haiqiong YU ; Zhixiong LIN ; Lidan MA ; Daozhong ZHU ; Zhiqiang GAO ; Xiaowei WU ; Yufeng TAO
Chinese Journal of Comparative Medicine 2024;34(6):93-99
Japan is one of the main trading partners for the import and export of experimental animals in China,and its quarantine and supervision policies for the import and export of experimental animals are very detailed and strict.This article takes experimental dogs,cats,and monkeys as examples to provide an in-depth analysis of the quarantine and supervision policies for the main experimental animals exported to Japan.At the same time,it reflects on the current laws and regulations,import and export management method,standards,biosafety,breeding and management status,as well as the import and export business status of experimental animals in China.Suggestions are provided in improving the laws and regulations,import and export management method,ensuring national biosafety,improving the management level of experimental animal breeding,and promoting the import and export trade of experimental animals,in order to provide reference for comprehensively improving the production,use,and breeding management level of experimental animals in China and strengthening the trade between China and Japan.
3.Effect and mechanism of dioscin on renal injury in septic rats
Xiang SHEN ; Shanggang XU ; Yanghui HUANG ; Bin LUO ; Yufeng ZHOU ; Longbin LIANG
China Pharmacy 2024;35(11):1334-1338
OBJECTIVE To investigate the effect of dioscin on renal injury in septic rats and its possible mechanism. METHODS The septic rat model was induced by using cecal ligation and puncture. Sixty model rats were randomly divided into model group (0.5% sodium carboxymethyl cellulose solution), dioscin low-dose, medium-dose and high-dose groups (30, 60, 120 mg/kg) and dexamethasone group (positive control, 10 mg/kg), with 12 rats per group; another 12 rats were selected as the sham operation group (0.5% sodium carboxymethyl cellulose solution). After 15 minutes of modeling, rats in each group were injected with medicine/0.5% sodium carboxymethyl cellulose solution via the tail vein. Twenty-four hours after administration, the levels of creatinine (Cr), blood urea nitrogen (BUN), neutrophil gelatinase associated lipocalin (NGAL), kidney injury molecule-1 (KIM- 1), interleukin 6 (IL-6), IL-1β and tumor necrosis factor-α (TNF-α) in serum and malondialdehyde (MDA) in renal tissue, superoxide dismutase (SOD) activity and the protein expressions of nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), NOD-like receptor protein 3 (NLRP3) were detected; renal histomorphology was observed. RESULTS Compared with model group, pathological injury of renal tissue was improved significantly in dioscin low-dose, medium-dose and high-dose groups; the levels of Cr, BUN, NGAL, KIM-1, IL-6, IL-1β and TNF-α in serum, MDA level and protein expression of NLRP3 in renal tissue were decreased significantly (P<0.05); SOD activity in renal tissue, protein expressions of Nrf2 and HO-1 were increased significantly (P<0.05), in a dose-dependent manner (P<0.05). The pathological damage of renal tissue in the dioscin high-dose group was similar to dexamethasone group, and there was no statistically significant difference in the levels of the above indicators (P>0.05). CONCLUSIONS Dioscin can activate the Nrf2/HO-1 signaling pathway to inhibit NLRP3 inflammasome, and realize the inhibition of inflammatory factors and oxidative stress, so as to protect the kidney injury in sepsis.
4.Effect and mechanism of dioscin on renal injury in septic rats
Xiang SHEN ; Shanggang XU ; Yanghui HUANG ; Bin LUO ; Yufeng ZHOU ; Longbin LIANG
China Pharmacy 2024;35(11):1334-1338
OBJECTIVE To investigate the effect of dioscin on renal injury in septic rats and its possible mechanism. METHODS The septic rat model was induced by using cecal ligation and puncture. Sixty model rats were randomly divided into model group (0.5% sodium carboxymethyl cellulose solution), dioscin low-dose, medium-dose and high-dose groups (30, 60, 120 mg/kg) and dexamethasone group (positive control, 10 mg/kg), with 12 rats per group; another 12 rats were selected as the sham operation group (0.5% sodium carboxymethyl cellulose solution). After 15 minutes of modeling, rats in each group were injected with medicine/0.5% sodium carboxymethyl cellulose solution via the tail vein. Twenty-four hours after administration, the levels of creatinine (Cr), blood urea nitrogen (BUN), neutrophil gelatinase associated lipocalin (NGAL), kidney injury molecule-1 (KIM- 1), interleukin 6 (IL-6), IL-1β and tumor necrosis factor-α (TNF-α) in serum and malondialdehyde (MDA) in renal tissue, superoxide dismutase (SOD) activity and the protein expressions of nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), NOD-like receptor protein 3 (NLRP3) were detected; renal histomorphology was observed. RESULTS Compared with model group, pathological injury of renal tissue was improved significantly in dioscin low-dose, medium-dose and high-dose groups; the levels of Cr, BUN, NGAL, KIM-1, IL-6, IL-1β and TNF-α in serum, MDA level and protein expression of NLRP3 in renal tissue were decreased significantly (P<0.05); SOD activity in renal tissue, protein expressions of Nrf2 and HO-1 were increased significantly (P<0.05), in a dose-dependent manner (P<0.05). The pathological damage of renal tissue in the dioscin high-dose group was similar to dexamethasone group, and there was no statistically significant difference in the levels of the above indicators (P>0.05). CONCLUSIONS Dioscin can activate the Nrf2/HO-1 signaling pathway to inhibit NLRP3 inflammasome, and realize the inhibition of inflammatory factors and oxidative stress, so as to protect the kidney injury in sepsis.
5.Surgical correction of true plus-minus lid syndrome: a case report
Xinxin LI ; Yufeng LIU ; Ning LIU ; Liang HU
Chinese Journal of Plastic Surgery 2024;40(7):743-746
True plus-minus lid syndrome is a rare clinical phenomenon characterized by ptosis on one side of the upper eyelid and uncompensated retraction on the other side. The reported cases in the past were all acquired plus-minus lid syndrome cases. This article presented a 26-year-old female patient with true plus-minus lid syndrome, who was in good health with no history of other underlying diseases, surgery or trauma. Her upper eyelid had been asymmetrical since childhood, showing retraction on the right and mild ptosis on the left, and it was left untreated. In March 2022, the patient was admitted to the Department of Plastic Surgery at the Second Hospital of Nanjing, Nanjing University of Chinese Medicine. The levator muscle of the upper eyelid was shortened on the left side, and the levator muscle of the upper eyelid was released and lengthened on the right side by surgery. After a 17-month follow-up, the bilateral corneal exposure rate was basically symmetrical, and the operative effect was satisfactory. The author also discussed the symptoms, signs, and differential diagnosis of true and pseudo plus-minus lid syndrome by reviewing the literature and inferred the pathogenesis of this case, which is helpful for selecting surgical methods.
6.Surgical correction of true plus-minus lid syndrome: a case report
Xinxin LI ; Yufeng LIU ; Ning LIU ; Liang HU
Chinese Journal of Plastic Surgery 2024;40(7):743-746
True plus-minus lid syndrome is a rare clinical phenomenon characterized by ptosis on one side of the upper eyelid and uncompensated retraction on the other side. The reported cases in the past were all acquired plus-minus lid syndrome cases. This article presented a 26-year-old female patient with true plus-minus lid syndrome, who was in good health with no history of other underlying diseases, surgery or trauma. Her upper eyelid had been asymmetrical since childhood, showing retraction on the right and mild ptosis on the left, and it was left untreated. In March 2022, the patient was admitted to the Department of Plastic Surgery at the Second Hospital of Nanjing, Nanjing University of Chinese Medicine. The levator muscle of the upper eyelid was shortened on the left side, and the levator muscle of the upper eyelid was released and lengthened on the right side by surgery. After a 17-month follow-up, the bilateral corneal exposure rate was basically symmetrical, and the operative effect was satisfactory. The author also discussed the symptoms, signs, and differential diagnosis of true and pseudo plus-minus lid syndrome by reviewing the literature and inferred the pathogenesis of this case, which is helpful for selecting surgical methods.
7.Study on the Effect of Qiai Essential Oil Promotes the Healing of Infectious Skin Wounds by Activating the PI3K/Akt Signa-ling Pathway
Feng LYU ; Yunjing LIANG ; Wei SONG ; Yufeng YAO ; Zongguo HONG
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(8):795-803
OBJECTIVE To observe the effect of Qiai(Artemisia argyi Levl.et Van.var.argyi cv.Qiai)essential oil on the healing of infected skin wounds and explore its mechanism.METHODS 30 Kunming mice were randomly divided into five equal groups:blank control group,substrate group,positive control group(3 mL·kg-1 Mupirocin ointment),low-dose group(each patch containing 7.3 mg of Qiai essential oil),and high-dose group(each patch containing 57.6 mg of Qiai essential oil).After anesthe-sia,a 10 mm full-thickness circular skin wound was created on the back of each mouse.Following hemostasis,100 μL of Staphylococ-cus aureus bacterial solution(1.5×108 CFU·mL-1)was applied to the wound.The wounds in each group were treated in the afore-mentioned manner,the dressing was changed daily,and the healing of the skin wound was observed.On the 12th day post-surgery,the mice were euthanized and the skin around the wounds was collected for HE and Masson staining to observe pathological features;qPCR and Western blotting were used to detect the expression of genes related to wound healing(Col1a1,Col3a1,Fn1,VEGFA),in-flammatory factors(IL-1β,IL-6,TNF-α),and the PI3K/Akt signaling pathway in the wound skin tissue.Further,in NIH-3T3 mouse embryonic fibroblasts,the potential impact of Artemisia argyi volatile oil on PI3K/Akt signaling pathway activity was investigated using the PI3K inhibitor PI-103.RESULTS Animal experiments have shown that compared to the blank control group,the relative wound area in both the low-dose and high-dose groups significantly decreased;the infiltration of inflammatory cells in the wound tis-sues reduced,while the expression of collagen fibers increased;qPCR showed that,compared to the blank control group,the expres-sion of VEGFA,Col3a1,and Col1a1 in the low-dose and high-dose groups significantly increased,while the expression of inflamma-tory factors IL-1β and TNF-α decreased;further Western blotting revealed that compared to the blank control group,both the phos-phorylated PI3K/total PI3K ratio and the phosphorylated Akt/total Akt ratio in the low-dose and high-dose groups significantly in-creased in a dose-dependent manner.Cellular experimental results also confirmed that the volatile oil from Artemisia argyi exerts a reg-ulatory effect on the PI3K/Akt signaling pathway in NIH-3T3 cells.CONCLUSION The essential oil of Qiai can promote wound healing,with its mechanism possibly involving the activation of the PI3K/Akt signaling pathway,suppression of inflammatory factor ex-pression,and promotion of collagen expression.
8.Macrophage ATG16L1 expression suppresses metabolic dysfunction-associated steatohepatitis progression by promoting lipophagy
Qi WANG ; Qingfa BU ; Zibo XU ; Yuan LIANG ; Jinren ZHOU ; Yufeng PAN ; Haoming ZHOU ; Ling LU
Clinical and Molecular Hepatology 2024;30(3):515-538
Background/Aims:
Metabolic dysfunction-associated steatohepatitis (MASH) is an unmet clinical challenge due to the rapid increased occurrence but lacking approved drugs. Autophagy-related protein 16-like 1 (ATG16L1) plays an important role in the process of autophagy, which is indispensable for proper biogenesis of the autophagosome, but its role in modulating macrophage-related inflammation and metabolism during MASH has not been documented. Here, we aimed to elucidate the role of ATG16L1 in the progression of MASH.
Methods:
Expression analysis was performed with liver samples from human and mice. MASH models were induced in myeloid-specific Atg16l1-deficient and myeloid-specific Atg16l1-overexpressed mice by high-fat and high-cholesterol diet or methionine- and choline-deficient diet to explore the function and mechanism of macrophage ATG16L1 in MASH.
Results:
Macrophage-specific Atg16l1 knockout exacerbated MASH and inhibited energy expenditure, whereas macrophage-specific Atg16l1 transgenic overexpression attenuated MASH and promotes energy expenditure. Mechanistically, Atg16l1 knockout inhibited macrophage lipophagy, thereby suppressing macrophage β-oxidation and decreasing the production of 4-hydroxynonenal, which further inhibited stimulator of interferon genes(STING) carbonylation. STING palmitoylation was enhanced, STING trafficking from the endoplasmic reticulum to the Golgi was promoted, and downstream STING signaling was activated, promoting proinflammatory and profibrotic cytokines secretion, resulting in hepatic steatosis and hepatic stellate cells activation. Moreover, Atg16l1-deficiency enhanced macrophage phagosome ability but inhibited lysosome formation, engulfing mtDNA released by pyroptotic hepatocytes. Increased mtDNA promoted cGAS/STING signaling activation. Moreover, pharmacological promotion of ATG16L1 substantially blocked MASH progression.
Conclusions
ATG16L1 suppresses MASH progression by maintaining macrophage lipophagy, restraining liver inflammation, and may be a promising therapeutic target for MASH management.
9.Research progress in the treatment of primary insomnia with electroacupuncture
Liang GUO ; Yufeng BI ; Changchun JI ; Zhangyin SUN ; Hui SUN ; Yuhan XUE
International Journal of Traditional Chinese Medicine 2023;45(3):381-384,F4
Electroacupuncture treatment of primary insomnia is widely used and with confirmed efficacy. The factors influencing the therapeutic effect of electroacupuncture include waveform selection, treatment frequency, stimulation intensity, stimulation time and acupoint selection. The mechanism of electroacupuncture in treating this disease mainly includes regulating the levels of excitatory neurotransmitters, γ-aminobutyric acid, melatonin and interleukin cytokines.
10.Treatment of tibial plateau fractures using modified double reverse traction technique
Hongjun CHEN ; Baijun HU ; Dawei GAO ; Yufeng WU ; Huahui LIANG ; Fukai GAN ; Huankun LI ; Gang QIN
Chinese Journal of Orthopaedic Trauma 2023;25(4):351-355
Objective:To study the clinical efficacy of our modified double reverse traction technique in the treatment of tibial plateau fractures.Methods:A retrospective study was conducted of the 66 patients with tibial plateau fracture who had been treated by our modified double reverse traction technique at Articular Department, Zhongshan Hospital of Traditional Chinese Medicine from March 2019 to December 2021. There were 37 males and 29 females, with an age of (44±14) years. The double reverse traction technique was used in all patients for reduction of the tibial plateau fracture, and their collapsed articular surfaces were restored by our self-designed universal high affinity prying technique, or by fibular graft support, or by the hooping technique to restore the transverse diameter of the tibial plateau which had been widened. The outcomes of fracture reduction at 2 days postoperation were recorded and evaluated by Rasmussen imaging assessment. Complications were also recorded. The visual analog scale (VAS) pain scores and Hospital for Special Surgery (HSS) scores were compared between preoperation, 3 and 6 months postoperation and the rates of fracture healing between 3 and 6 months postoperation.Results:This group of 66 patients was followed up for 12.0(8.0, 16.5) months. The Rasmussen imaging assessment at 2 days postoperation: 60 excellent, 4 good, and 2 fair cases, yielding an excellent to good rate of 97.0% (64/66). At 3 and 6 months postoperation, the VAS scores [1 (0, 1) and 0 (0, 1)] and HSS scores [84 (78, 88) and 91 (85, 95)] were significantly improved compared with those before operation [4 (3, 5) and 36 (29, 39)], and the values at 6 months postoperation were significantly improved compared with those at 3 months postoperation ( P<0.05). The rate of clinical fracture healing at 6 months postoperation (100%, 66/66) was significantly higher than that at 3 months postoperation (77.3%, 51/66) ( P<0.05). Perioperatively, no adverse events were observed like incision infection, deep vein thrombosis of lower limbs, or failure or exposure of internal fixation, except for delayed incision healing in only one patient. Conclusion:In the minimally invasive surgery for tibial plateau fractures, our modified double reverse traction technique can result in excellent fracture reduction by imaging assessment, fine functional recovery of the knee joint and relief of pain.

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