ObjectiveTo investigate the effects of modified Buyang Huanwu Tang on cerebral ischemia-reperfusion injury （CI/RI） in mice via the PTEN-induced putative kinase 1/E3 ubiquitin ligase （PINK1/Parkin） signaling pathway-mediated mitophagy， and to explore the underlying mechanism by which modified Buyang Huanwu Tang improves CI/RI. MethodsSeventy-two male C57BL/6J mice were randomly divided into six groups （n = 12 per group）： Sham-operated group， middle cerebral artery occlusion/reperfusion （MCAO/R） model group， low-， medium-， and high-dose modified Buyang Huanwu Tang groups （8.84， 17.68， 35.36 g·kg^-1·d^-1）， and an aspirin group （13.00 mg·kg^-1·d^-1）. Neurological deficit scores were assessed using the Zea-Longa method. Cerebral infarct volume ratio was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Histopathological changes and neuronal injury in brain tissues were observed using hematoxylin-eosin （HE） staining and Nissl staining. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） assay. Mitochondrial ultrastructure in brain tissue was observed by transmission electron microscopy （TEM）. Serum levels of superoxide dismutase （SOD） and malondialdehyde （MDA） were determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expression levels of PINK1， Parkin， microtubule-associated protein 1 light chain 3B （LC3B， LC3Ⅱ/Ⅰ）， and p62 in brain tissues were detected by real-time quantitative reverse transcription PCR （Real-time PCR） and Western blot， respectively. ResultsCompared with the sham-operated group， the MCAO/R model group showed significantly increased neurological deficit scores and cerebral infarct volume ratios （P<0.01）. Severe cortical injury on the infarct side was observed， characterized by decreased neuronal density， cytoplasmic vacuolation， nuclear pyknosis， a marked reduction in Nissl bodies， dissolution of Nissl bodies in the cytoplasm of some pyramidal neurons， and blurred cellular boundaries. The number of TUNEL-positive cells increased significantly （P<0.01）. Mitochondria exhibited cristae membrane rupture and matrix vacuolation， with rupture of the outer mitochondrial membrane and formation of autophagosomes， the number of which increased significantly. Serum SOD activity decreased significantly （P<0.01）， while MDA content increased significantly （P<0.01）. In infarcted brain tissues of model mice， the relative mRNA expression and protein levels of PINK1， Parkin and LC3B were significantly increased （P<0.05， P<0.01）， whereas p62 mRNA and protein expression were significantly decreased （P<0.05， P<0.01）， showing statistical significance. Compared with the model group， all treatment groups showed significantly decreased neurological deficit scores and cerebral infarct volume ratios （P<0.01）. Neuronal density increased significantly， cytoplasmic vacuolation was alleviated， nuclear morphology tended to be more regular and clearer， Nissl body density increased significantly with reduced dissolution and improved contour clarity. The mitochondrial cristae structure was partially restored， with some mitochondria showing autophagosome encapsulation， and the degree of mitochondrial damage was alleviated. Serum SOD activity increased significantly （P<0.01）， while MDA content decreased significantly. The mRNA and protein expression levels of PINK1， Parkin， and LC3Ⅱ/Ⅰ were significantly increased （P<0.05， P<0.01）， while p62 mRNA and protein expression in the low- and medium-dose modified Buyang Huanwu Tang groups were significantly decreased （P<0.05， P<0.01）， showing statistical significance. ConclusionModified Buyang Huanwu Tang can upregulate the protein expression levels of PINK1， Parkin， and LC3Ⅱ/Ⅰ and downregulate p62 protein expression， suggesting that it may improve CI/RI by regulating the expression of proteins related to the PINK1/Parkin signaling pathway. Regulation of the mitophagy pathway may be one of the mechanisms by which modified Buyang Huanwu Tang alleviates CI/RI in mice.

Objective To investigate the epidemiological characteristics of cross-county imported dengue fever cases within Yunnan province in 2023, so as to provide insights into formulation of preventive and control measures for intra-provincial spread of dengue fever. Methods All data pertaining cross-county imported dengue fever cases within Yunnan Province in 2023 were collected, and the temporal, regional and population distributions of the cases were descriptively analyzed. Results A total of 1 664 intra-provincial cross-county imported dengue fever cases were reported in 95 counties (cities, districts) cross 16 profectures (cities) in Yunnan Province in 2023, accounting for 12.34% of total cases in the province. Cross-county imported dengue fever cases were predominantly reported during the period between August and October (1 516 cases, 91.11% of total cases), and peaked in September (659 cases), with a single-day peak on October 8 (36 cases). During the period from September 4 to 10, five counties (cities) with local dengue fever epidemics, including Jinghong City of Xishuangbanna Dai Autonomous Prefecture, Gengma Dai and Wa Autonomous County of Lincang City, Ruili City of Dehong Dai and Jingpo Autonomous Prefecture, Mengla Coun ty of Xishuangbanna Dai Autonomous Prefecture, and Zhenkang County of Lincang City, exported 165 cross-county imported dengue fever cases to the rest of the province. Among the 1 644 intra-provincial cross-county imported dengue fever cases, the male to female ratio was 1.40∶1.00, and 1 329 cases were at ages of 15 to 55 years (79.87%), with farmers as the predominant occupation (886 cases, 53.25%). The top 5 counties (cities/districts) reporting the highest number of intra-provincial cross-county imported dengue fever cases included Simao District (266 cases) and Lancang Lahu Autonomous County (118 cases) of Pu’er City, Mengla County (91 cases) and Menghai County (91 cases) of Xishuangbanna Dai Autonomous Prefecture, and Mangshi City (73 cases) of Dehong Dai and Jingpo Autonomous Prefecture, which accounting for 38.40% of total imported cases. These intra-provincial cross-county imported dengue fever cases originated from 7 counties (cities/districts) in 4 prefectures (cities), including 1 261 cases (76.70%) from Jinghong City of Xishuangbanna Dai Autonomous Prefecture, 224 cases (13.63%) from Ruili City of Dehong Dai and Jingpo Autonomous Prefecture, 103 cases (6.27%) from Gengma Dai and Wa Autonomous County of Lincang City, 31 cases (1.89%) from Mengla County of Xishuangbanna Dai Autonomous Prefecture, 30 cases (1.82%) from Zhenkang County of Lincang City, 10 cases (0.61%) from Cangyuan Wa Autonomous County of Lincang City, and 5 cases (0.30%) from Mohan-Boten Economic Cooperation Zone of Kunming City. In addition, local dengue fever epidemics following intra-provincial cross-county importation of dengue fevers cases in Simao District, Jinggu Dai and Yi Autonomous County, Mangshi City, Longchuan County, and Cangyuan Wa Autonomous County. Conclusions Farmers and students are high-risk populations for intra-provincial cross-county imported dengue fever cases in Yunnan Province, and health education pertaining personal protection against dengue fever should be strengthened among these high-risk populations by governments at all levels. There is a high risk of local out-break of dengue fever following continuous introduction of intra-provincial cross-county imported cases. Standardized management of intra-provincial cross-county imported dengue fever cases should be reinforced to reduce the risk of local epidemics.

Objective To explore the effect of the knockdown of transmembrane 9 superfamily protein member 2 (TM9SF2) on the replication of vesicular stomatitis virus (VSV), and investigate its role in the mechanism of antiviral innate immunity. Methods Small interfering RNA (siRNA) was used to knock down the TM9SF2 gene in human non-small cell lung cancer A549 cells. The CCK-8 method was used to assess cell proliferation. A VSV-green fluorescent protein (VSV-GFP) infected cell model was established. The plaque assay was used to measure the viral titer in the supernatant. RT-qPCR and Western blotting were employed to quantify the mRNA and protein levels of VSV genome replication in A549 cells following VSV infection, as well as the expression of interferon β (IFN-β) mRNA and interferon regulatory factor 3 (IRF3) protein phosphorylation following polyinosinic-polycytidylic acid (poly(I:C)) stimulation. Results Compared to the negative control, the knockdown of TM9SF2 exhibited a significant effect, with no observed impact on A549 cell proliferation. The VSV-GFP infected A549 cell model was successfully established. After viral stimulation, fluorescence intensity was reduced following TM9SF2 knockdown, and the mRNA and protein levels of VSV were significantly downregulated. The viral titer of VSV was decreased. After poly(I:C) stimulation, TM9SF2 knockdown significantly upregulated the mRNA level of IFN-β and the phosphorylation level of IRF3 protein. Conclusion The knockdown of TM9SF2 inhibits the replication of vesicular stomatitis virus, and positively regulates the type I interferon signaling pathway, thus enhancing the host's antiviral innate immune response.
Humans ; Virus Replication/genetics* ; Signal Transduction ; Membrane Proteins/metabolism* ; A549 Cells ; Vesiculovirus/physiology* ; Interferon-beta/metabolism* ; Interferon Regulatory Factor-3/genetics* ; Interferon Type I/metabolism* ; Vesicular Stomatitis/immunology* ; Gene Knockdown Techniques ; Vesicular stomatitis Indiana virus/physiology* ; RNA, Small Interfering/genetics*

Flap surgery is a complex surgical procedure that has become an effective method for the treatment of many diseases and traumas.Flap survival is closely related to a variety of factors including cellular autophagy,oxidative stress,inflammatory response,mesenchymal stem cells(MSCs)function,and vascular regeneration.Cellular autophagy maintains intracellular homeostasis and plays a key role in reducing oxidative stress and inflammation and promoting injury repair.Excessive oxidative stress and inflammatory responses pose a threat to flaps,affecting their survival and successful transplantation.Endothelial cells are involved in vascu-lar regeneration through proliferation,migration,and production of angiogenic factors,and vascular endothelial growth factor directly promotes blood vessel formation and maintains endothelial cell function.MSCs play an important role in promoting flap survival and tissue repair due to their unique biological properties and multiple mechanisms of action.The multiple roles played by cellular autophagy,oxidative stress,inflammatory response,MSCs function,and vascular regeneration in influencing postoperative flap survival are hereby elaborated.The aim is to provide a basis for the clinical application of regulating the above factors to improve postoperative flap survival,improve the success rate of flap surgery,reduce complications,and bring more hope for the recovery and quality of life of patients.

[Objective]To investigate the protective effect and mechanism of Ganoderma lucidum extract(GLE)on liver cirrhosis in mice.[Methods]Ten male C57BL/6 mice were randomly selected as control group,the remaining forty mice were intraperitoneally injected with carbon tetrachloride olive oil suspension to induce liver cirrhosis model.They were randomly divided into model group and GLE low(50 mg/kg·d),medium(100 mg/kg·d)and high(200 mg/kg·d)dose groups,while the control group and model group received 0.9％sodium chloride solution gastric irrigation,and the control group mice were given the same volume of olive oil solution twice a week.Liver index was calculated.The activities of alanine aminotransferase(ALT),aspartate transaminase(AST)and the levels of total cholesterol(TC),total bilirubin(TB)and creatinine(Cr)in serum of mice were detected by automatic biochemical analyzer.Hematoxylin eosin(HE)staining was used to observe the histopathological changes of liver,and Masson staining was used to observe the degree of liver fibrosis.Terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL)staining was used to observe the apoptosis of hepatocytes.The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6,malondialdehyde(MDA)and activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in serum were detected by enzyme linked immunosorbent assay(ELISA).The relative expression of nuclear factor E2 related factor 2(Nrf2)and nuclear Nrf2,heme oxygenase-1(HO-1)and NAD(P)H:quinone oxidoreductase 1(NQO1),α-smooth muscle actin(α-SMA),Collagen Ⅰ and E-cadherin protein in liver tissue were detected by Western blot.[Results]Compared with control group,the liver had significant damage,the liver index,serum ALT,AST activities,TC,TB and Cr levels,liver fibrosis degree,hepatocyte apoptosis index,the levels of serum TNF-α,IL-1 β,IL-6 and MDA,the relative expression of α-SMA and Collagen I protein increased(P＜0.05),while the activity levels of serum SOD and GSH-Px,and the relative expression of total Nrf2 and nuclear Nrf2,HO-1,NQO1 and E-cadherin protein in liver tissue decreased in model group(P＜0.05).Compared with model group,liver injury gradually reduced,the liver index,serum ALT,AST activities,TC,TB and Cr levels,liver fibrosis degree,hepatocyte apoptosis index,the levels of serum TNF-α,IL-1β,IL-6,MDA and the relative expression of α-SMA,Collagen I protein decreased(P＜0.05),while the activity levels of serum SOD and GSH-Px,and the relative expression of Nrf2 and nuclear Nrf2,HO-1,NQO1 and E-cadherin protein in liver tissue increased in GLE low,medium and high dose groups(P＜0.05).[Conclusion]GLE can alleviate the histopathological damage and improve liver function in cirrhotic mice.This may be related to the decreased level of oxidative stress and inflammatory reaction after activation of Nrf2/ARE signaling pathway,which may interfere with liver fibrosis.

Objective:To investigate the early auditory discrimination of vowels, consonants and lexical tones in prelingually-deafened children with cochlear implants (CI) using auditory event-related potentials.Methods:Nineteen prelingually-deafened CI children and 19 normal hearing (NH) children were recruited in this study. A multi-deviant oddball paradigm was constructed using the monosyllable/ta1/as the standard stimulus and monosyllables/tu1/,/te1/, /da1/,/ra1/,/ta4/and/ta2/as the deviant stimuli. The event-related potentials evoked by vowel, consonant and lexical tone contrasts were recorded and analyzed in the two groups.Results:NH children showed robust mismatch negativities (MMNs) to vowel, consonant and lexical tone contrasts ( P<0.05), whereas CI children only showed positive mismatch responses (pMMRs) and P3a responses to the vowel ( P<0.05) and consonant contrasts ( P<0.05) and no significant event-related potential to the lexical tone contrasts ( P>0.05). The longer pMMR and P3a peak latencies ( P<0.01) but similar amplitudes ( P>0.05) were found in CI children than in NH children. CI children showed weaker phase synchronization of θ oscillations than NH children ( P<0.05). The duration of CI use was positively correlated with the scores of Categories of Auditory Performance (CAP) ( P=0.004), Speech Intelligibility Rate (SIR) ( P=0.044) and Meaningful Auditory Integration Scale (MAIS) ( P=0.001) in CI children. Conclusions:Prelingually-deafened CI children can process vowels and consonants at an early stage. However, their ability of processing speech, especially lexical tones, is still more immature compared with their NH peers. The event-related potentials could be objective electrophysiological indicators reflecting the maturity of CI children′s auditory speech functions. Long-term CI use is beneficial for prelingually-deafened children to improve auditory and speech performance.

Flap surgery is a complex surgical procedure that has become an effective method for the treatment of many diseases and traumas. Flap survival is closely related to a variety of factors including cellular autophagy, oxidative stress, inflammatory response, mesenchymal stem cells (MSCs) function, and vascular regeneration. Cellular autophagy maintains intracellular homeostasis and plays a key role in reducing oxidative stress and inflammation and promoting injury repair. Excessive oxidative stress and inflammatory responses pose a threat to flaps, affecting their survival and successful transplantation. Endothelial cells are involved in vascular regeneration through proliferation, migration, and production of angiogenic factors, and vascular endothelial growth factor directly promotes blood vessel formation and maintains endothelial cell function.MSCs play an important role in promoting flap survival and tissue repair due to their unique biological properties and multiple mechanisms of action. The multiple roles played by cellular autophagy, oxidative stress, inflammatory response, MSCs function, and vascular regeneration in influencing postoperative flap survival are hereby elaborated. The aim is to provide a basis for the clinical application of regulating the above factors to improve postoperative flap survival, improve the success rate of flap surgery, reduce complications, and bring more hope for the recovery and quality of life of patients.

OBJECTIVE To establish the fingerprint of Guiqi tongmai mixture and evaluate its quality in combination with chemical pattern recognition . METHODS Using narirutin as reference peak ，HPLC fingerprint of Guiqi tongmai mixture was established with Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 edition）. Common peaks identification and similarity evaluation were conducted . Hierarchical cluster analysis （HCA），principle component analysis （PCA）and orthogonal partial least square -discriminate analysis （OPLS-DA）were performed ，and the value of variable importance in the projection （VIP） greater than 1 was taken as the standard to screen the differential markers affecting the quality of Guiqi tongmai mixture . RESULTS Nineteen common peaks in the HPLC fingerprints of thirteen batches of Guiqi tongmai mixture were calibrated ，and the similarities of them were 0.928-0.990；eight common peaks were identified ，including the peak 7（hydroxysafflor yellow A ），peak 8（albiflorin），peak 9（paeoniflorin），peak 10（calycosin-7-O-glucoside），peak 12（liquiritin），peak 14（ammonium glycyrrhizinate ）， peak 16（narirutin）and peak 19（hesperidin）. According to the results of HCA ，thirteen batches of samples could be clustered into three categories ：S5，S9 and S 11-S13 were clustered into one category ；S4 and S 8 were clustered into one category ；S1-S3，S6- S7 and S 10 were clustered into one category . The results of PCA indicated that the accumulative variance contribution rate of the principle components 1-7 was 92.115%. The results of OPLS -DA showed that VIP values of narirutin ，paeoniflorin，liquiritin， ammonium glycyrrhizinate ，the components corresponding to peak 11 and peak 1 were greater than 1. CONCLUSIONS HPLC fingerprint of Guiqi tongmai mixture is established ，and it could be used for quality evaluation of Guiqi tongmai mixture by combining with chemical pattern recognition techniques . Six components such as narirutin are differential markers affecti ng the quality of Guiqi tongmai mixture .

Liangmianzhen(Zanthoxyli Radix) has long been used as medicine. The current medicinal parts are different from those in the ancient. As recorded in the Chinese Pharmacopeia, the medicinal part is root. However, in ancient works, the medicinal parts include root, stem, leaf, and fruit. In an attempt to find the historical basis that stem is a reasonable medicinal part, the herbalogical study was carried out on this medicinal based on the formal names, synonyms, original plant, medicinal parts, habitat of the medicinal plant, producing area, processing and preparation methods, efficacy, and indications recorded in ancient Chinese materia medica and local gazetteers. The results showed that Liangmianzhen was firstly recorded as a medicinal in Shennong's Classic of Materia Medica with the formal name of "Manjiao". "Manjiao" was adopted from the Han Dynasty to the Qing Dynasty when it was changed to "Rudijinniu", the name originating from the folk in the south of the Five Ridges. Now, the formal name is "Liangmianzhen", which was firstly recorded in Wuxuan County Gazetteer in 1914 and then as a synonym in the Updated Records of Picking Herbs in the South of the Five Ridges. According to the formal names, synonyms, and the descriptions of the original plant, the medicinal plants of Liang-mianzhen have the characteristics of shrub-like young seedlings, vine adult seedlings, corymbiform thyrsus, stems with thorns, amphitropous golden-yellow roots with horn-like branches, and thorns on both sides of the leaves. Thus, "Manjiao", "Rudijinniu", and "Liangmianzhen" were from the same species of Zanthoxylum nitidum(Rutaceae), which was also verified based on the growth environment, habitat, processing and preparation methods, efficacy, and indications. In ancient times, the stem and root were the main medicinal parts and leaves and fruits were also used. However, in the Chinese Pharmacopeia, root is recorded as the only medicinal part, which is obviously inconsistent with the records in the ancient classics. In light of the limited medicinal resources for Liang-mianzhen, other medicinal parts of Z. nitidum is recommended. This study clarified the medicinal parts of Z. nitidum in history. It is recommended that the stem be added to the medicinal parts of Z. nitidum in the next edition of Chinese Pharmacopeia.
China ; Drugs, Chinese Herbal ; Fruit ; Materia Medica ; Medicine, Chinese Traditional ; Plants, Medicinal

The aim of this study was to investigate the effect of transmembrane 9 superfamily protein member 2 (TM9SF2) in proliferation and migration of triple negative breast cancer cell line MDA-MB-231.The expression of TM9SF2 in triple negative breast cancer cell line MDA-MB-231 and nontumorigenic mammary epithelial cell line MCF-10A were measured by Western blot. MDA-MB-231 cells were treated with siRNA-TM9SF2 to knock-down the expression of TM9SF2. The effect of silencing TM9SF2 was measured with Western blot.The proliferation of cells was tested by MTS,and the migration was measured with Transwell and wound-healing assay.Proteins related to proliferation (PI3K,AKT,SRC and ERK) and migration (Snail,Slug and N-cadherin) were measured with Western blot.Protein expressions of TM9SF2 was better improved in triple negative breast cancer MDA-MB-231 cell line than MCF-10A.Compared with the control group, the siRNA-TM9SF2 infected group had lower expressions of PI3K, Snail, Slug and N-cadherin, and at the same time phosphorylation of AKT was decreased. The results suggest TM9SF2 can promote the proliferation and metastasis of triple negative breast cancer MDA-MB-231 cell line.