The genomic island(GI)characteristics and virulence factor differences of clinical isolates of Burkholderia pseudomallei in Hainan Province,China were analyzed to provide a scientific basis for diagnosis and treatment of melioidosis.In total,52 B.pseudomallei isolates were collected for detection of virulence-related GIs by PCR.The whole genome sequence annotation format file was submitted on Islandviwer 4 platform,and the genomes of the same species and close relatives were added for comparison.Two algorithms,SIGI-HMM and IslandPath-DIMOB,were integrated to predict GIs and sequence a-lignments were conducted to identify specific GIs and differences in virulence factors.The genomes of 52 clinical strains could be divided into three branches based on evolutionary distance,with 82.69％(43/52)of strains concentrated in branch 1.In to-tal,828 GIs were identified among the 52 B.pseudomallei genomes,which formed 157 GI clusters based on sequence similari-ty.GIs accounted for 2.05％-6.38％of the genome content.While GI clusters 1 and 2 were present in all strains,a total of 84(53.50％)GI clusters only clustered within a single genome isolate.Of 10 GI likely specific clusters,five were from the same genus,two from another genus,and three with uncertain origins.Moreover,25 GI clusters were associated with virulence,which included eight shared by B.pseudomallei BP76 and BP169,which had the highest number of virulence-associated GIs among all isolates.O the 52 B.pseudomallei isolates,variations were identified in the virulence genes fhaB1,fhaB2,BPSL1661,cheY1,wzM,tssH-5/clpV,tssA-5,boaA,and boaB.Comparisons of these findings with clinical isolates from Thailand and Australia showed that B.pseudomallei isolates from Hainan had significant differences in the sequences of boaA,boaB,cheY1,and chbp.Additionally,fhaB1,fhaB3,and bimA displayed significant variations specifically within the Australian isolates.B.pseudomallei GI was conserved and specific to Hainan.The identification of specific GI and virulence factors was useful to clarify the source of horizontal gene transfer and differences in virulence at the molecular level.

Objective:To investigate the effect of GRIM-19 on the resistance of carcinoma cells to the chemotherapeutic agent docetaxel in the treatment of PCa.Methods:Using siRNA technology to interfere with the gene expression in PCa cells,we estab-lished a model of GRIM-19 overexpression/knockdown in PCa cells.We investigated the effect of different expression levels of GRIM-19 on docetaxel-induced death of the PCa cells by qPCR,Western blot and flow cytometry,and assessed the value of GRIM-19 in re-ducing the chemotherapy-resistance of PCa cells.Results:GRIM-19 was down-regulated in PCa tissues and cells.Knockout of GRIM-19 significantly decreased the expression of siGRIM19 in the PC-3 and LNCaP cells,and reduced their death rate when treated with docetaxel compared with the control group.The expressions of GRIM-19 mRNA and protein were remarkably upregulated after transfection with GRIM-19,and the overexpressed GRIM-19 promoted the death of the PC-3 and LNCaP cells treated with docetaxel in a dose-dependent manner.Flow cytometry analysis showed a lower apoptosis rate of PC-3-R cells than that of PC-3 cells at different time points of docetaxel-induction at different doses.Conclusion:GRIM-19 is a PCa suppressor gene with a significant facilitating effect on the apoptosis of PCa cells,and the overexpression of GRIM-19 promotes docetaxel-induced PCa cell death and improves the sensitivity of chemotherapy.

Objective:To study the mechanism of icariin inhibiting the proliferation of human PCa PC-3 cells based on cell metabolomics technology.Methods:We determined the proliferation activity of human PC-3 cells by methyl thiazolyl tetrazolium(MTT)assay,and compared the proliferation of the PC-3 cells among the control,5-fluorouracil and icariin intervention groups.Using the Bligh Dyer method,we extracted endogenous metabolites from the cells,analyzed the metabolic profile by ultra-high pressure liquid chromatography tandem quadrupole time-of-flight mass spectrometry,identified the differential metabolites by principal component anal-ysis and orthogonal partial least-squares discrimination analysis,and enriched the metabolic pathways based on the MetaboAnalyst data-base.Results:Icariin significantly inhibited the proliferation of human PCa PC-3 cells.A total of 89 differential metabolites were i-dentified,mainly including amino acids,phosphatidylcholine,phosphatidylethanolamine,lysophosphatidylcholine,and lysophosphati-dylethanolamine,all with the tendency to return to the normal level after icariin intervention.Icariin significantly downregulated the metabolic levels of the glycerophospholipid metabolites phosphatidylcholine,phosphatidylethanolamine,lysophosphatidylcholine and ly-sophosphatidylethanolamine,and upregulated those of amino acid metabolites tryptophan,leucine,and proline in the PC-3 cells.Conclusion:Icariin inhibits the proliferation of human PCa PC-3 cells,which may be closely related to its regulatory effect on lipid metabolism(glycerophospholipid metabolism)and amino acid metabolism.

AIM To investigate the mechanism of triterpenoids quillaic acid and gypsogenin-3-O-glucuronide from Psammosilene tunicoides on tunicamycin-induced rheumatoid arthritis fibroblasts-like synoviocytes(RA-FLS)via the endoplasmic reticulum pathway.METHODS The research objects of tunicamycin-induced RA-FLS intervened with quillaic acid and gypsogenin-3-O-glucuronide had their cell proliferation activity detected;their level of tumor nerosis factor-α(TNF-α)detected by ELISA;their apoptosis detected by flow cytometry;their cell migration ability detected by Transwell experiment;their expressions of transcription activator 6(ATF-6),glucose regulatory protein 78(GRP78),C/EBP homologous protein(CHOP),cysteine protease protein-12(caspase-12)and anti-apoptosis Bcl-2 protein detected by Western blot;and their mRNA expressions of ATF-6,GRP78 and CHOP detected by RT-qPCR.RESULTS Compared with the model group,each group intervened with quillaic acid or gypsogenin-3-O-glucuronide displayed decreased levels of TNF-α(P＜0.01);weakened cell proliferation and migration ability(P＜0.01);increased apoptosis rate(P＜0.01);decreased protein expressions of ATF-6 and Bcl-2(P＜0.05,P＜0.01);and increased protein expressions of CHOP and caspase-12(P＜0.05,P＜0.01).In addition,decreased GRP78 protein expression in the low and medium dose groups(P＜0.05,P＜0.01);decreased mRNA expression of ATF-6,GRP78(P＜0.01)and increased CHOP mRNA expression(P＜0.01)in the medium dose groups of quillaic acid and gypsogenin-3-O-glucuronide were observed as well.CONCLUSION Quillaic acid and gypsogenin-3-O-glucuronide may play a protective role in rheumatoid arthritis by inhibiting the proliferation and migration of RA-FLS,inducing apoptosis and reducing the secretion of related inflammatory factors via endoplasmic reticulum signal pathway.

AIM To evaluate the quality of Yanyangke Mixture.METHODS The HPLC fingerprints were established,after which cluster analysis,principal component analysis and partial least squares discriminant analysis were performed.The contents of liquiritin,rosmarinic acid,sheganoside,irisgenin,honokiol,monoammonium glycyrrhizinate,irisflorentin,isoliquiritin and magnolol were determined,the analysis was performed on a 35 ℃ thermostatic Agilent ZORBAX SB-C18 column(5 μm,250 mmx4.6 mm),with the mobile phase comprising of 0.1％phosphoric acid-acetonitrile flowing at 1 mL/min in a gradient elution manner,and multi-wavelength detection was adopted.RESULTS There were ten common peaks in the fingerprints for twelve batches of samples with the similarities of more than 0.9.Various batches of samples were clustered into three types,three principal components displayed the acumulative variance contribution rate of 87.448％,peaks 5、14(honokiol),3(liquiritin),11(monoammonium glycyrrhizinate)and 15(asarinin)were quality markers.Nine constituents showed good linear relationships within their own ranges(r＞0.999 0),whose average recoveries were 98.5％-103.6％with the RSDs of 0.92％-1.7％.CONCLUSION This stable and reliable method can provide a basis for the quality control of Yanyangke Mixture.

Objective:To analyze the etiology and risk factors of Zang nationality cerebral infarction patients of different ages in Xizang Autonomous Region, so as to provide basis for more targeted diagnosis, treatment and prevention of cerebral infarction in this region.Methods:The clinical data of 500 Zang nationality cerebral infarction patients hospitalized in Xizang Autonomous Region People′s Hospital from January 2019 to December 2023 were retrospectively analyzed. According to age, they were divided into young and middle-aged group (18-59 years old) and elderly group (60-75 years old). Baseline data, laboratory data and imaging results of patients in each group were collected and retrospectively analyzed.Results:The proportion of males in the young and middle-aged group ( n=267) [188 (70.41%)] was higher than that in the elderly group ( n=233) [130 (55.79%), χ 2=11.485, P=0.001]. The proportion of smokers [131 (49.06%) vs 74 (31.76%), χ 2=15.401], drinkers [121 (45.32%) vs 84 (36.05%), χ 2=4.417], high altitude polycythemia (HAPC)[ 51 (19.10%) vs 23 (9.87%), χ 2=8.406], hyperuricemia (HUA)[ 61 (28.90%) vs 34 (19.32%), χ 2=4.766], increased hemoglobin [152 (56.93%) vs 97 (41.63%), χ 2=6.677], hypertriglyceridemia [47 (17.60%) vs 18 (7.73%), χ 2=10.734], hypercholesterolemia [12 (4.94%) vs 3 (1.29%), χ 2=4.397], hyperlipidemia [79 (29.59%) vs 43 (18.45%), χ 2=8.360] in the young and middle-aged group was higher than that in the elderly group (all P<0.05). The proportion of hypertension [108 (40.44%) vs 158 (67.81%), χ 2=37.413], atrial fibrillation [5 (1.87%) vs 20 (8.58%), χ 2=11.797], hyperhomocysteinemia (HHcy)[159 (59.55%) vs 168 (72.10%), χ 2=8.664], abnormal creatinine [18 (6.74%) vs 29 (12.45%), χ 2=4.755], atherosclerosis [113 (42.32%) vs 145 (62.23%), χ 2=19.748], heart disease [135 (50.56%) vs 150 (64.38%), χ 2=9.690] in the young and middle-aged group was lower than that in the elderly group (all P<0.05). Multivariate Logistic regression analysis showed that hypertension ( OR=2.865, 95% CI 1.742-4.710) and HHcy ( OR=1.968, 95% CI 1.177-3.290) were risk factors of cerebral infarction in the elderly group. Smoking ( OR=1.848, 95% CI 1.017-3.360), HAPC ( OR=1.993, 95% CI 1.991-4.011), HUA ( OR=1.863, 95% CI 1.015-3.419) and living at the extremely high altitude ( OR=2.405, 95% CI 1.207-4.791) were risk factors of cerebral infarction in the young and middle-aged group. According to the TOAST etiological classification, the causes of stroke of other determined etiology and stroke of other undetermined etiology were complex and diverse, which were more common in the young and middle-aged group, while cardiac embolism was more common in the elderly group. Conclusions:There are significant differences in the risk factors and etiology of cerebral infarction in different age groups in Xizang Autonomous Region. The occurrence of cerebral infarction in elderly patients is significantly related to hypertension and HHcy, while the occurrence of cerebral infarction in young and middle-aged patients is significantly related to smoking, HAPC, HUA, and living in extremely high altitude areas. In clinical practice, diagnosis and treatment of cerebral infarction patients in different age groups should have different focuses.

Objective:To investigate the application value of transanal intersphincteric resec-tion under direct vision in the Jackknife position in the anal preserving surgery for ultra-low rectal cancer.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 15 patients with ultra-low rectal cancer who underwent transanal intersphincteric resection under direct vision in the Jackknife position, combined with laparoscopic total mesorectal excision in Huizhou Municipal Central Hospital from September 2021 to November 2022 were collected. There were 9 males and 6 females, aged (63±9)years. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Surgical and postoperative outcomes. All the 15 patients underwent operations successfully, without conversion to open abdo-minal operation. There were 5 cases of partial ISR and 10 cases of subtotal ISR. The operation time and volume of intraoperative blood loss of 15 patients were (260±30)minutes and 20(range, 10-30)mL, respectively. The distance from anastomosis to anal margin was (1.6±0.8)cm. The duration of post-operative hospital stay was 10(range, 8-13)days, and all 15 patients underwent colonic and anasto-mosis with staplers and protective ileostomy at the terminal ileum. Three patients had postoperative complications within 30 days after surgery, of whom 1 case with grade A anastomotic leakage was cured after conservative treatment and 2 cases with anastomotic membranous stenosis were cured by anal enlargement. (2) Postoperative pathological examination. The number of lymph nodes dissected of 15 patients was 18 ±6, and the distance between the tumor and distal resection margin was 1.3(range, 1.0-2.0)cm. The surgical specimens of all 15 patients showed complete mesorectum and negative for proximal, distal and circumferential margins. Results of postoperative pathological examination showed that there was 1 case in stage pT1N0M0, 9 cases in stage pT2N0M0, 1 case in stage pT2N1M0, 1 case in stage ypT0N0M0, 2 cases in stage ypT2N0M0, 1 case in ypT3N1M0 stage. The histological subtype showed 11 cases of moderately differentiated adenocarcinoma and 4 cases of well-differentiated adenocarcinoma. (3) Follow-up. All 15 patients were followed up for 15(range, 12-24)months. No local recurrence and distant metastasis of the tumor was found, and no tumor-related death occurred. All 15 patients underwent stoma closure. The postoperative anal function assessment of 15 patients showed no disorder in 5 cases, mild disorder in 8 cases and severe disorder in 2 cases.Conclusion:Transanal intersphincteric resection under direct vision in the Jackknife position in the anal preserving surgery for ultra-low rectal cancer is safe and feasible.

One patient was admitted to a hospital due to"sepsis,chronic kidney disease,type 2 diabetes,shock,and cerebral infarction".Patient's blood specimen was taken for clinical examination.Aerobic and anaerobic culture results of catheter blood and venous blood were both positive.The pathogen was identified as Staphylococcus pas-teuri by VITEK MS,and the patient was diagnosed as catheter-related bloodstream infection caused by Staphylo-coccus pasteuri.Clinical empirical use of piperacillin for anti-infection treatment was ineffective,and vancomycin was eventually used for treatment based on in vitro antimicrobial susceptibility testing.Patient's condition improved after removing the venous catheter.There are currently no reported cases of Staphylococcus pasteuri in China.Ear-ly identification of pathogen and adjustment of treatment plans based on antimicrobial susceptibility testing results are crucial for effective treatment of this case.

ObjectivesTo understand the prevalence of hyperhomocysteinemia （HHcy） in the Tibetan population in Northwest Xizang， and its association with lipids and blood uric acid， and to explore the prevention and treatment strategies for chronic diseases such as HHcy in Northwest Xizang. MethodsIn this survey， questionnaires， physical examinations， and biochemical tests were conducted on 3432 Tibetan residents aged 18 years and older who had resided in Northwest Xizang （Ngari Prefecture， Nagqu City） for more than 6 months using a multistage stratified whole cluster random sampling method. ResultsThe prevalence of HHcy among Tibetan residents in Northwest Xizang was 75.7%， much higher than that of the average population in China （37.2%）. Blood uric acid、high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were risk factors for HHcy. ConclusionThe prevalence of HHcy is higher in the Tibetan population in northwest Xizang. Therefore， the local governments should urge people to establish a healthy lifestyle and enhance early intervention for HHcy by improving diet and lifestyle， thereby reducing the risk of cardiovascular， cerebrovascular and other related diseases.

OBJECTIVE: To investigate whether electroacupuncture (EA) at sensitized acupoints could reduce sympathetic-sensory coupling (SSC) and neurogenic inflammatory response by interfering with 5-hydroxytryptamine (5-HT)ergic neural pathways to relieve colitis and somatic referred pain, and explore the underlying mechanisms. METHODS: Rats were treated with 5% dextran sodium sulfate (DSS) solution for 7 days to establish a colitis model. Twelve rats were randomly divided into the control and model groups according to a random number table (n=6). According to the "Research on Rat Acupoint Atlas", sensitized acupoints and non-sensitized acupoints were determined. Rats were randomly divided into the control, model, Zusanli-EA (ST 36), Dachangshu-EA (BL 25), and Xinshu (BL 15) groups (n=6), as well as the control, model, EA, and EA + GR113808 (a 5-HT inhibitor) groups (n=6). The rats in the control group received no treatment. Acupuncture was administered on 2 days after modeling using the stimulation pavameters: 1 mA, 2 Hz, for 30 min, with sparse and dense waves, for 14 consecutive days. GR113808 was injected into the tail vein at 5 mg/kg before EA for 10 min for 7 consecutive days. Mechanical sensitivity was assessed with von Frey filaments. Body weight and disease activity index (DAI) scores of rats were determined. Hematoxylin and eosin staining was performed to observe colon histopathology. SSC was analyzed by immunofluorescence staining. Immunohistochemical staining was performed to detect 5-HT and substance P (SP) expressions. The calcitonin gene-related peptide (CGRP) in skin tissue and tyrosine hydroxylase (TH) protein levels in DRG were detected by Western blot. The levels of hyaluronic acid (HA), bradykinin (BK), prostaglandin I2 (PGI2) in skin tissue, 5-HT, tryptophan hydroxylase 1 (TPH1), serotonin transporters (SERT), 5-HT 3 receptor (5-HT3R), and 5-HT 4 receptor (5-HT4R) in colon tissue were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: BL 25 and ST 36 acupoints were determined as sensitized acupoints, and BL 15 acupoint was used as a non-sensitized acupoint. EA at sensitized acupoints improved the DAI score, increased mechanical withdrawal thresholds, and alleviated colonic pathological damage of rats. EA at sensitized acupoints reduced SSC structures and decreased TH and CGRP expression levels (P<0.05). Furthermore, EA at sensitized acupoints reduced BK, PGI2, 5-HT, 5-HT3R and TPH1 levels, and increased HA, 5-HT4R and SERT levels in colitis rats (P<0.05). GR113808 treatment diminished the protective effect of EA at sensitized acupoints in colitis rats (P<0.05). CONCLUSION EA at sensitized acupoints alleviated DSS-induced somatic referred pain in colitis rats by interfering with 5-HTergic neural pathway, and reducing SSC inflammatory response.
Rats ; Animals ; Electroacupuncture ; Rats, Sprague-Dawley ; Serotonin ; Acupuncture Points ; Pain, Referred ; Calcitonin Gene-Related Peptide ; Signal Transduction ; Colitis/therapy* ; Indoles ; Sulfonamides