Six compounds were isolated from the ethyl acetate fraction of Citri Sarcodactylis Fructus by using various column chromatographic methods, such as MCI Gel CHP-20, ODS, Sephadex LH-20, silica gel and semipreparative HPLC. Their structures were identified to be citrusin G (1), citrusin H (2), citrusin E (3), syringin (4), coniferin (5), methylconiferin (6) by NMR, HR-ESI-MS, UV, IR spectra. Compounds 1 and 2 were new secondary metabolism products and 3-6 were obtained from the titled material for the first time. Compound 1 showed anti-renal fibrosis activity in TGF-β1-induced kidney proximal tubular cells.

Objective To investigate the current status and serologic characteristics of occult HBV infection in the adult physical examination population in Zigong region. Methods A total of 126 381 patients who were examined in the physical examination center and gastroenterology department of The First People's Hospital of Zigong City from April 2023 to September 2024 were screened, and 21 615 eligible cases were included in the study. The current status of infection was analyzed and serological patterns and serological characteristics of the included individuals were compared. Results This study screened 126 381 patients, all of whom underwent serum HBsAg testing, and 21 615 patients (17.10%) underwent HBV DNA testing, of which 7 992 were HBV DNA positive (>102 IU/mL) and HBsAg negative, accounting for 36.97% of the total number of patients who underwent HBV DNA testing. Anti-HBc positivity was significantly higher than other serologic patterns, and the lowest rate of HBV DNA positivity was found in those who were positive for anti-HBc, anti-HBs and anti-HBe. The lowest male-to-female ratio (1.25:1) was found in patients with both anti-HBc, anti-HBs and anti-HBe positivity, which was significantly lower than that of patients with the other three serologic characteristics (P=0.005). There were no significant differences in age, BMI, AST, ALT, and TBiL levels among patients with different serum characteristics (all P>0.05). The HBV viral load is highest in patients with anti HBc combined with anti HBe positivity, while the HBV viral load is lowest in patients with anti HBc positivity, anti HBs positivity, and all anti HBe positivity (P<0.001). Viral genotypes were predominantly B-type, and there were differences in genotype distribution among the four groups of patients (P<0.001). Conclusion The level of occult HBV infection was high in the adult medical examination population in Zigong region, mostly characterized by anti-HBc positivity, with the lowest male-to-female ratio among patients who were positive for anti-HBc, anti-HBs, and anti-HBe, and the highest HBV viral load among patients who were positive for anti-HBc combined with anti-HBe.

Aim To explore the effect of total fla-vonoids of Rhodiola japonica(TFR)on vascular forma-tion in rat cerebral vascular endothelial cells in vitro and its relationship with VEGFR2 and neurogenic hy-drogen sulfide(H2 S).Methods Using rat cerebral vascular endothelial cells cultured separately and co cultured with hippocampal neurons,different experi-mental methods were used to detect cell proliferation,migration,tube formation,H2S content,and calcium fluorescence intensity,including CCK8 method,cell scratch method,Transwell method,matrix gel tube for-mation,H2 S reagent kit,and calcium fluorescence probe method.Results In isolated cultured rat brain microvascular endothelial cells,H2S donor NaHS(200μmol·L-1)and TFR(90,270,810 mg·L-1)could significantly promote the proliferation,migration area and the number of migrating cells of primary rat brain microvascular endothelial cells,as well as the forma-tion of blood vessels and[Ca2+]i,while VEGFR2 blocker SU5416(10 μmol·L-1)could inhibit the promotion of endothelial cell proliferation,migration and vascular formation by TFR;TFR significantly in-creased H2S content in co-cultured rat brain microvas-cular endothelial cells with hippocampal neurons,and was inhibited by CBS inhibitor AOAA(200 μmol· L-1).At the same time,TFR significantly promotes the formation of vascular effects in co-cultured rat cere-bral vascular endothelial cells,and could be signifi-cantly inhibited by AOAA and VEGFR2 blocker SU5416.Conclusions TFR can promote the forma-tion of blood vessels in cerebral vascular endothelial cells by increasing[Ca2+]i of VEGFR2 in vitro,and can promote angiogenesis by inducing CBS generation of H2S in neurons and acting on VEGFR2 in rat brain microvascular endothelial cells.

Objective To analyze the efficacy of two-hole and four-hole laparoscopic surgery in patients with sigmoid carcinoma,and to explore the influencing factors leading to complications.Methods The clinical data of 130 patients with sigmoid carcinoma who underwent laparoscopic surgery in our hospital from October 2020 to December 2022 were analyzed.The patients were divided into groups according to the surgical perforation methods.Sixty-eight patients in two-hole group were treated with two-hole laparoscopic surgery and 62 patients in four-hole group were treated with four-hole laparoscopic surgery.The perioperative indexes,visual analogue scale(VAS)scores and postoperative complications of patients in the two groups were observed and compared,and the influencing factors of complications were analyzed.Results The total incision length,first time to get out of bed,first postoperative exhaust time,postoperative hospital stay and VAS score at each time point after surgery of patients in two-hole group were significantly shorter/lower than those in four-hole group(P<0.05).There was no statistically significant difference in the incidence of complications between the two groups(P>0.05).Univariate analysis showed that there were statistically significant differences in the age,preoperative albumin,operation time,intraoperative blood loss,first time to get out of bed,first postoperative exhaust time,postoperative hospital stay,VAS scores at the 3rd day and 5th days after surgery of patients in the occurrence group and the non-occurrence group(P<0.05).Further Logistic regression analysis showed that operation time and preoperative albumin<35 g/L were the main influencing factors for the occurrence of complications(P<0.05).Conclusion Compared with four-hole laparoscopic surgery,two-hole laparoscopic surgery for patients with sigmoid carcinoma has shorter time of perioperative recovery,less postoperative pain,and no difference was found in postoperative complications between the two methods.Operation time and preoperative albumin level<35 g/L were the main influencing factors for the occurence of complications.

To evaluate the effectiveness and safety of implantable D-shant atrial shunt device in patients with severe pulmonary arterial hypertension(PAH)and right heart failure.A 53-year-old female patient diagnosed with severe idiopathic PAH and right heart failure,her WHO FC grade was Ⅳ.The right heart catheter and implantation of D-shant atrial shunt device were performed under local anesthesia on November 30,2021.A 6 mm×4 cm peripheral artery balloon was selected to dilate the atrial septum and a D-shant atrial shunt device with a fixed 4 mm diameter orifice was implanted into the heart.The clinical symptoms and hemodynamics of the patient was improved after the intervention.Implantation of atrial shunt device as a palliative therapy to established a right to left shunt is another strategy for treating patients with severe PAH in late period,which has good effectiveness and safety.It could be the last replacement therapy to improve symptoms and prolonged lives to drug resistant and severe PAH patients.

This study was aimed at exploring the epidemiological characteristics of plague,and the evolutionary relation-ships among the isolated plague strains in the Yunnan border area,to provide clues for further studying epidemic causes and ep-idemiological patterns.Plague epidemic data in the border area during the second epidemic period(1982-2007)were collected and analyzed with descriptive epidemiological methods.Whole genome sequences of 262 strains of Yersinia pestis in the border area were obtained for phylogenetic analysis.Plague outbreaks occurred in 17 counties(cities)among 25 border counties(cit-ies);a total of 552 epidemic foci and 123 human cases were identified.The 1.ORI2,1.ORI3,1.IN3,2.ANT and 2.MED geno-types were identified among Yersinia pestis isolated from the Yunnan border area,among which the 1.ORI2 population was dominant.A total of 258 strains of Yersinia pestis from the 1.OR12 population belonged to four subclusters.The Myanmar and Vietnam clade was embedded within the Yunnan clade in the overall phylogeny.The above results indicated that during the sec-ond period of the epidemic,the intensity of plague epidemics in Yunnan's border areas was high,showing a trend of devel-opment from west to south and east.Our findings indicated a risk of cross-border transmission of plague between Yunnan and neighboring countries;therefore,the surveillance,pre-vention,and control of plague in border areas should be strengthened.

Exosomes can ameliorate neuronal cell injury induced by hypoxia-ischemia,but the relation-ship between astrocyte-derived exosomes(As-exo)and mitochondrial function,mitochondrial associated ER membrane(MAM)function and whether mitochondrial autophagy is relevant is currently unclear.The aim of this study was to investigate the role of astrocyte-derived exosomes in the regulation of mito-chondrial function,MAM and mitochondrial autophagy in PC 12 cells after oxygen and glucose depriva-tion/reoxygenation(OGD/R).Exosomes were extracted from the supernatant of the astrocyte culture me-dium by ultracentrifugation.Using the live cell imaging system,we observed that fluorescently labeled exosomes could show obvious enrichment in PC 12 cells at 24 h.Meanwhile,co-localization of exosomes with mitochondria could be observed under the laser confocal scanning microscope;mitochondrial pres-sure changes were detected using the Seahorse cellular energy metabolism fractionation instrument.The result showed that basal respiration in the OGD/R group,compared with that in the control group,proton leakage,maximal respiration and ATP-related respiration were significantly reduced(P＜0.05 or P＜0.01),and all four indexes were elevated and statistically significant in the OGD/R+exo group compared with the control group(P＜0.05 or P＜0.01).The results of the co-localization of the mitochondria and ER showed that the structure of the MAM was harmed by oxygen-sugar deprivation and then reoxygen-ation,and the structure of As-exo and the mitochondria appeared to have a distance-reduced polymeriza-tion phenomenon,while the mitochondria and ER co-localized.The co-localization results of mitochondri-a and ER showed that the structure of MAM was damaged by oxygen deprivation and reoxygenation,and the aggregation phenomenon of MAM was weakened by the treatment of As-exo;the flow-through results showed that As-exo could restore the decrease of the mitochondrial membrane potential and the elevation of the ROS by oxygen deprivation to a certain degree.Western blotting showed that As-exo could signifi-cantly inhibit the mitochondrial autophagy-associated tension protein homologue induced hypothetical ki-nase 1(PTEN induced kinase 1(PINK1)and Parkin protein(parkin RBR E3 ubiquitin protein ligase(Parkin))were elevated,and the addition of As-exo decreased LC3 Ⅱ/LC3 Ⅰ protein expression,ele-vated P62 protein expression,and reduced OGD/R-induced mitochondrial autophagy.The results showed that OGD/R treatment can cause mitochondrial dysfunction,MAM structural changes and increased mito-chondrial autophagy in PC12 cells,and As-exo treatment can improve mitochondrial function,attenuate the formation of MAM,and reduce mitochondrial autophagy in PC 12 cells,which can have the potential of preventing the reperfusion injury in ischemic stroke.

As a unique gene in the genome,FLASH(FADD-like interleukin-1β-converting enzyme asso-ciated huge protein)/CASP8AP2 is involved in multiple cellular processes,including apoptosis,histone gene pre-mRNA processing,transcriptional regulation,and cell cycle progression.Clinical studies have shown that FLASH is a valuable prognostic marker for acute lymphoblastic leukemia,and a crucial factor for the survival of various cancer cells.Therefore,in-depth research into the function of FLASH may offer new perspectives for the treatment of related diseases.Our previous research identified FLASH as a bind-ing partner of p53,demonstrating that FLASH enhances the transcriptional activity of p53.Here we fur-ther investigate the molecular mechanisms of the interaction between FLASH and p53,revealing that the p53-K386R mutation(SUMOylation residue)attenuated its interaction with FLASH(aa 51-200)and FLASH-SIM(SUMO-interacting motif)(aa 1 534-1 806)significantly.However,SUMO can bind to FLASH-SIM directly,instead of FLASH(aa 51-200).Subsequent research shows that overexpression of FLASH in cells enhances global SUMO1 conjugation and p53-SUMO1 conjugation,therefore providing a plausible explanation for the underlying mechanism of FLASH enhancing the transcriptional activity of p53.Since promyelocytic leukemia protein nuclear body(PML NB)serves as subcellular reactors for SUMO conjugation within the cell,and the PML Ⅳ isoform can specifically enhance the SUMO modifica-tion of p53,we have investigated the interaction between FLASH and PML Ⅳ,and elucidated the struc-tural basis of their interaction:both FLASH-N3A(501-802)and FLASH-C2(1 807-1 981)bind to PML Ⅳ(aa 228-633).Further investigations reveal that they can synergistically enhance global SUMO1 modification as well as SUMO1 modification of p53.The interaction between FLASH and tumor suppres-sors p53 or PML Ⅳ enriches our understanding of its function and reveals the potential mechanism of FLASH in tumor development,therefore offering novel insights into cancer diagnosis and treatment.

Objective:To investigate the accuracy of next-generation sequencing technology(NGS)in detecting the polymorphisms of HLA-DRB1,DQB1,DQA1,DRB3,DRB4,DRB5,DPA1 and DPB1 alleles in randomly-selected unrelated healthy individuals from Shenzhen Han population,investigate the potential reason for HLA-DRB1 allele dropout in routine NGS,and establish an internal quality control system.Methods:NGS-based HLA class Ⅱ genotyping was performed on 1 012 samples using the MiSeqDxTM platform.The suspected missed alleles indicated by the quality control software and HLA-DRB1 homozygotes were confirmed by PCR-SSOP or PCR-SBT methods.Results:A total of 139 alleles were detected,including HLA-DRB1(45),DRB3(7),DRB4(5),DRB5(7),DQA1(17),DQB1(21),DPA1(10)and DPB1(27).HLA-DRB 1*09:01(17.09％),15:01(10.72％);DRB3*02:02(25.99％),03:01(10.18％);DRB4*01:03(36.46％);DRB5*01:01(15.42％);DQA1*01:02(20.01％),03:02(17.19％);DQB1*03:01(19.47％),03:03(17.98％),05:02(11.66％),06:01(10.67％);DPA1*02:02(54.45％),01:03(31.18％)and DPB1*05:01(39.13％),02:01(16.90％)alleles were the most common alleles in Shenzhen Han population(frequencies＞10％).There was no statistical difference between the gene frequencies of HLA-DRB1 and DQB1 loci in our study.The HLA Common and Well-Documented Alleles in China(CWD2.4)(x2=12.68,P＞0.05).94 cases of HLA-DRB1 homozygous samples detected by NGS were retested by PCR-SSOP or SBT method,and one case of allele dropout at HLA-DRB1 locus was found.SBT method confirmed that the allele of DRB1*04:03 was missed.The laboratory internal quality control system was established.Two cases of new alleles were detected and named by WHO Nomenclature Committee for Factors of the HLA System.Conclusion:The HLA genotyping results based on NGS showed a significantly lower ambiguity rate.The HLA class Ⅱ alleles exhibit genetic polymorphism in the Han population of unrelated healthy individuals in Shenzhen.The independent method based on NGS in clinical histocompatibility testing has limitations and requires internal quality control strategies to avoid allele-dropout events.

Allyl isothiocyanate (AITC) is a natural product commonly used in food preservation and pharmaceutical applications. Toxoplasmosis, caused by the protozoan pathogen Toxoplasma gondii, is prevalent globally while the impact of AITC on toxoplasmosis is unclear. We explored the effect of AITC on acute toxoplasmosis. We infected C57BL/6 mice with T. gondii type I RH strain following AITC administration. On the 4th day after infection, which corresponds to the initial stage of infection, we collected serum for the determination of inflammatory cytokine levels. The mice serum of the AITC-administered group contained significantly lower levels of granulocyte colony-stimulating factor, interferon-gamma, interleukin (IL)-23 subunit p19, IL-4, IL-6, and monocyte chemoattractant protein-1. The lifespan of the mice in the AITC-administered group was significantly reduced. In vitro experiments showed that AITC promoted the proliferation of intracellular T. gondii accompanied by the inhibition of IL-4, IL-1β, and IL-6 production in RAW264.7 macrophages. Our results showed that AITC facilitated T. gondii infection in the early stage by inhibiting the production of several inflammatory cytokines.