OBJECTIVE To explore the effect of volatile oil of Ligusticum chuanxiong on the transdermal properties and cytotoxicity of triptolide in vitro. METHODS The chemical constituents of the volatile oil of L. chuanxiong were analyzed by gas chromatography-mass spectrometry. The lower abdominal skin of KM mice was separated and divided into triptolide group， triptolide in compatibility with volatile oil of L. chuanxiong groups at 1∶10， 1∶50， 1∶100 （hereinafter referred to as “compatibility 1∶10”“compatibility 1∶50”“compatibility 1∶100” groups）. After the skin of mice in each group was fully exposed to 0.2 g of the corresponding cream for 24 h， the cumulative transdermal dose （Qn） of triptolide in the receiving solution of each group was determined by high-performance liquid chromatography， and the transdermal absorption rate （Jss） was calculated. Human immortalized keratinocytes （HaCat） were used as a model， the CCK-8 method was used to detect the cell survival rate of different concentrations of the volatile oil of L. chuanxiong and triptolide before and after compatibility. RESULTS A total of 62 chemical constituents of the volatile oil of L. chuanxiong were identified， including Z-ligustilide， senkyunolide， and β-selinene. The Qn （P＜ 0.01） and Jss of triptolide increased within 24 h in the compatibility 1∶10 and 1∶50 groups， while the Qn （P＜0.05） and Jss decreased in the compatibility 1∶100 group as compared with the triptolide group. Compared with the triptolide group， the cell survival rate of HaCat was significantly increased in the compatibility 1∶10 and 1∶50 groups when the triptolide concentrations were 36， 72 and 144 ng/mL （P＜0.05 or P＜0.01）； while the cell survival rate of HaCat was decreased in the compatibility 1∶100 group， but the difference was not statistically significant （P＞0.05）. CONCLUSIONS When the compatibility ratio of triptolide and volatile oil of L. chuanxiong was 1∶10 or 1∶50， it can promote the transdermal absorption of triptolide and reduce the cytotoxicity of triptolide to HaCat.

Objective To investigate the role and molecular mechanism of 3,3′-diindolelmethane(DIM),a natural plant compound in cruciferous vegetables,in the prevention and treatment of gastric cancer,and then provide experimental evidence for the application of dietary plant nutrition in the prevention of gastric cancer.Methods After human gastric cancer cell line BGC-823 was treated with dif-ferent concentrations of DIM,including 0,20,40,60,80,100,and 120 μmol/L,for 24 hours,the cell survival rate was determined by the tetramethylazolium blue(MTT)method.Western blot was used to detect the expression levels of Calpain,activating transcrip-tion factor 4(ATF4)and CAAT/enhancer-binding protein homologous protein(CHOP).The changes of intracellular calcium ion con-centration were detected by a calcium ion fluorescent probe.After ATF4 in BGC-823 cells was specifically knocked down using the small interfering RNA(siRNA)of ATF4,the expression levels of ATF4 and CHOP were detected by Western blot and the cell survival rate was determined by the MTT method.After the BGC-823 cells were pretreated with the calcium ion chelator BAPTA-AM,the ex-pression levels of calpain,ATF4 and CHOP proteins were measured by Western blot and the changes in cell proliferation activity were determined by the MTT method.Results The detection results of the MTT,Western blot,and calcium ion fluorescent probe methods showed that with the increase of DIM concentration,the proliferation ability of BGC-823 cells significantly decreased(P<0.05).Mean-while,the expressions of Calpain,ATF4,and CHOP proteins were up-regulated(P<0.05)and the fluorescence intensity of intracellu-lar calcium ion increased.After transfection with the siRNA of ATF4,the expression level of ATF4 in BGC-823 cells was decreased(F=16.17,P<0.05).Compared with the DIM+si-NC group,the cell survival rate in the DIM+si-ATF4 group was increased(F=74.46,P<0.05).After the intracellular calcium ions were chelated by BAPTA-AM,the expression levels of Calpain,ATF4,and CHOP proteins in the DIM+BAPTA-AM group were significantly lower than that in the DIM group(F=24.58,34.93,and 24.45,re-spectively,P<0.05),while the cell survival rate was the opposite(F=31.13,P<0.05).Conclusion DIM can increase the concen-tration of intracellular calcium ions and up-regulate the expressions of Calpain,ATF4 and CHOP proteins,thereby inducing the endo-plasmic reticulum stress in gastric cancer cells and inhibiting their proliferation.

Objective:To investigate the potential source of infection for a cluster outbreak of human brucellosis in Yantai City, Shandong Province.Methods:The information of a human brucellosis cluster outbreak case in Yantai City, Shandong Province in 2022 was collected, the strains were isolated and cultured, and DNA was extracted. BCSP31-PCR was used for species identification, and AMOS-PCR was used for species type identification. Multiple locus variable-number tandem-repeat analysis (MLVA)-16 was used for clustering analysis, and the results were compared with the public database MLVAbank and the monitoring data of Brucella in Shandong Province in 2022. At the same time, whole genome single nucleotide polymorphism (wgSNP) typing was used to analyze the 53 Brucella strains that had completed whole genome sequencing in Shandong Province in 2022, and the wgSNP phylogenetic tree was constructed. Results:According to BCSP31-PCR and AMOS-PCR identification, the three strains related to the cluster outbreak of brucellosis in Yantai City, Shandong Province in 2022 were all Brucella melitensis biotype. The results of MLVA-16 typing showed that the MLVA-16 typing of the three isolated strains was completely consistent, with 16 tandem repeat loci of 1-5-3-13-2-2-3-2-4-41-8-4-4-3-6-5, belonging to the Eastern Mediterranean clade. Compared with MLVAbank, the MLVA-16 typing of two strains isolated from Kazakhstan was consistent with the results of this study. Compared with the monitoring data of Brucella in Shandong Province in 2022, it was found that the MLVA-16 typing of 11 isolated strains was consistent with the results of this study, which were isolated from Zaozhuang, Linyi, Taian, Yantai, and Weifang cities, respectively. The results of wgSNP typing showed that the distance between the 11 strains and the strains of the current outbreak was less than 7 single nucleotide polymorphisms, and the strains were isolated from Taian, Zibo, Linyi, Binzhou, Jinan, Jining, Yantai and Weihai cities, respectively. Conclusion:After tracing the source of a human brucellosis cluster outbreak in Yantai City, Shandong Province in 2022, it is speculated that the strains of Brucella melitensis isolated from Linyi, Taian and Yantai cities are closely related, indicating that sheep in these areas have homology.

Objective:To investigate the clinicopathological characteristics of dedifferentiated endometrial carcinoma/undifferentiated endometrial carcinoma (DDEC/UDEC) with loss of expression of SMARCA4.Methods:A total of 10 cases with loss of expression of SMARCA4 were diagnosed at Fujian Cancer Hospital between January 2019 and December 2023. A retrospective analysis was conducted on the clinical characteristics, morphology, immunophenotype, molecular classification, and prognosis.Results:(1) Clinical characteristics: among 10 cases of DDEC/UDEC with loss of expression of SMARCA4, the patients′ age ranged from 48 to 65 years, with a median age of 56 years.Five cases were classified as International Federation of Gynecology and Obstetrics (FIGO) stages Ⅰ-Ⅱ, while the remaining five were categorized as stages Ⅲ-Ⅳ. (2) Pathological features: tumor cells exhibited poor cell adhesion, with common intravascular tumor emboli (8/10), occasional vacuolated nuclei (6/10), rhabdoid cells (4/10), and starry sky phenomenon formed by tissue cell phagocytosis apoptosis bodies or fragments (4/10). Six cases (6/10) showed loss of mismatch repair (MMR) protein expression, two cases (2/10) exhibited p53 mutant expression, and five cases (5/10) tested positive for programmed cell death ligand 1 (PD-L1). (3) Molecular subtyping: molecular subtyping revealed POLEmut in 1 case (1/10), mismatch repair deficient (MMR-d) in 5 cases (5/10), p53 abn in 1 case (1/10), and no specific molecular profile (NSMP) in 3 cases (3/10). (4) Prognosis: the follow-up period ranged from 7 to 42 months, with a median of 20 months. Five patients succumbed to the tumor, whereas the remaining five exhibited no recurrence during subsequent postoperative evaluations. The 2-year progression-free survival rates and overall survival rates were 58.3% and 52.5%, respectively.Conclusions:Loss of expression of SMARCA4 occurs in approximately 1/5 of DDEC/UDEC, which presents with an aggressive clinical course and a poor prognosis. About half of them show MMR protein loss expression and PD-L1 positive expression, suggesting that there might be benefit from treatment with immune checkpoint inhibitors.

Objective:To investigate the role of speckle-type POZ(pox virus and zinc finger protein) protein (SPOP) in enterovirus 71 (EV71) infection.Methods:Immunoprecipitation analysis was employed to examine the impact of SPOP on the ubiquitin level of EV71 non-structural protein 2A protease (2A pro), while the phosphorylation level of IFR3 protein was assessed through Western blot. Cells were either overexpressed or knockdown of SPOP, followed by infection with EV71. RT-qPCR was utilized to analyze the transcription level of IFN-β, and the transcription level and protein level of EV71 structural protein VP1 were determined using RT-qPCR and Western blot, respectively. Results:The inhibition of EV71 infection in RD cells was observed following transfection with HA-SPOP. Additionally, it was found that the ubiquitin level of EV71-2A pro increased in a gradient-dependent manner. Subsequent transfection with shSPOP plasmid for endogenous SPOP knockdown resulted in a dose-dependent decrease in the levels of melanoma differentiation-associated gene 5 (MDA5), mitochondrial antiviral signaling (MAVS), and p-IRF3. Conversely, transfection with HA-SPOP plasmid led to a dose-dependent increase in the levels of MDA5, MAVS, and p-IRF3. The expression of SPOP, whether high or low, had an impact on the expression of IFN-β in cells. Additionally, the levels of VP1 mRNA or protein were found to be inhibited or increased. Conclusions:SPOP plays a role in increasing the ubiquitination level of EV71-2A pro, which in turn promotes the phosphorylation level of IRF3 and secretion of IFN-β. This effect is achieved by inhibiting the cleavage of 2A pro against key molecules MAVS and MDA5 in the RLR signaling pathway, ultimately leading to the inhibition of EV71 replication.

Objective:To investigate the risk factors of infection with polymyxin resistant and carbapenemase-resistant Klebsiella pneumoniae（PR-CRKP）. Methods:A total of 170 patients with CRKP infection admitted in the Third Affiliated Hospital of Soochow University from July 2020 to October 2023 were enrolled，including 123 cases of CRKP infection and 47 cases of PR-CRKP infection. The general conditions，exposure of antibacterial drugs 6 months before admission，laboratory test indicators at admission，antibacterial drug use when target bacteria were detected，length of hospital stay and time of invasive procedures in two groups were retrospectively analyzed. The risk factors of PR-CRKP infection were analyzed with univariate and multivariate logistic regression. SPSS 26.0 software was used to analyze the data.Results:Univariate analysis showed that compared with the CRKP group，the average age of patients in PR-CRKP group was older（ Z = -2.186， P = 0.029），the proportion of patients with exposure history to semisynthetic penicillins，carbapenems，polymyxins，and quinolones 6 months before admission was higher（ χ 2= 3.930，5.414，11.939，8.478，all P < 0.05），the proportion of infections diagnosed at admission and blood urea nitrogen levels（ χ 2= 7.268， Z = -2.406， P = 0.007 and 0.016）was higher，the hemoglobin level（ t = 2.641， P = 0.009）was lower，the length of hospital stay was longer，the rates of tracheal intubation，urinary catheter，and deep vein catheterization were higher（ Z = -4.243，-4.660，-5.341，-4.583，all P < 0.001），the duration of carbapenem and polymyxin B use was longer（ Z = -4.757，-7.554，both P < 0.001），the proportion of combined quinolone-resistant Escherichia coli（QREC）and carbapenem-resistant organism（CRO）infections and bloodstream infections，and the rate of admission to intensive care units was higher（ χ 2 = 33.737，42.041，5.426，12.991， P < 0.05 or < 0.01）. Multivariate analysis showed that time to polymyxin B use（ OR = 1.179， 95%CI 1.059-1.312， P = 0.003），combined QREC infection（ OR = 5.357， 95%CI 2.100-13.669， P < 0.001）and combined CRO infection（ OR = 3.302， 95%CI 1.146-9.514， P= 0.027）were independent risk factors for PR-CRKP. Conclusion:Prolonged use of polymyxin B is an independent risk factor for PR-CRKP，and mixed QREC and CRO infection can increase the risk of PR-CRKP.

Objectives Objectives To investigate how the imbalance of innate lymphoid cells (ILCs)in the peripheral blood of patients with lung adenocarcinoma affects the balance of downstream mononuclear macrophages and T helper (Th) cells, and to identify the impact of the imbalance of ILCs on the immune status and prognosis of lung adenocarcinoma. Methods The peripheral blood of 20 patients with lung adenocarcinoma and normal controls were collected. The percentage of ILCs, mononuclear macrophages and T lymphocyte in peripheral blood were analyzed by flow cytometry. The characteristic cytokine secretion levels of various types of immune cells in peripheral blood were detected by real-time fluorescence quantitative PCR. Results Compared with the normal controls, the proportion of M2 mononuclear macrophages, ILC1 and ILC2 in patients with lung adenocarcinoma was up-regulated, while the proportion of M1 mononuclear macrophages, CD4⁺ T and CD8⁺ T was down-regulated. The mRNA expression of related cytokines of M1 mononuclear macrophages and ILC1 were decreased; while the mRNA expression of related cytokines of M2 mononuclear macrophages and ILC2 were increased. Along with the decreased CD4⁺T cells-associated cytokine T-bet mRNA expression, and the increased GATA3 mRNA expression. Moreover, the expression of PD-1 in CD8⁺ T cells was also up-regulated. Conclusion The imbalance of ILCs in peripheral blood of patients with lung adenocarcinoma promotes the imbalance of mononuclear macrophages and Th cells, which altogether maintains the immunosuppression in patients with lung adenocarcinoma, and promotes the development of lung adenocarcinoma.
Humans ; Lymphocytes ; Immunity, Innate ; CD8-Positive T-Lymphocytes ; Cytokines/metabolism* ; Adenocarcinoma of Lung ; Immunosuppression Therapy ; RNA, Messenger

Ubiquitination modifications are a kind of post-translational modifications of proteins widely found in eukaryotes and involved in a variety of biological activities. E3 ubiquitin ligases are an important component of the ubiquitin system, with the function of specific recognition of substrate proteins and mediation of different types of ubiquitination modifications. They can regulate the function and life time of substrate proteins. Recent studies have shown that E3 ubiquitin ligases are widely involved in the regulation of the host innate immune response and can directly or indirectly influence viral infection. Moreover, viruses are able to encode or hijack E3 ubiquitin ligases in their long-term evolution, allowing them to play an important role in viral infection and replication cycle. This paper reviewed the progress in the mechanisms of E3 ubiquitin ligases in innate immune responses and viral infection in recent years.

Objective: To analyze the characteristics of individuals positive for SARS-CoV-2 nucleic acid in a centralized isolation site for people entering China in Huzhou City of Zhejiang Province from December 18, 2021 to January 12, 2022, so as to provide insights into the prevention and control of overseas imported COVID-19. Methods: The basic characteristics, nucleic acid detection and epidemiological investigations were collected from individuals positive for SARS-CoV-2 nucleic acid in a centralized isolation site for people entering China from December 18, 2021 to January 12, 2022, and the temporal distribution, population distribution, source of importation, and virus typing were descriptively analyzed. Results : From December 18, 2021 to January 12, 2022, a total of 2 974 individuals in 19 flights were recorded in this centralized isolation site, and 33 cases were tested positive for SARS-CoV-2 nucleic acid, including 21 confirmed cases with common type, 9 confirmed cases with mild type, and 3 cases with asymptomatic infections. There were 11 cases with Omicron infections ( 33.33% ), 5 cases with Delta infections ( 15.15% ), and 17 cases with infection of unidentified types ( 51.52% ). The median interval ( interquartile range ) from the time of entry to the time of a positive test was 4.0 ( 7.0 ) days among all positive cases, 0 ( 4.0 ) day among cases with Omicron infections and 4.5 ( 8.5 ) days among cases with infections of Delta and unidentified types. The positive cases had a mean age of ( 36.97±8.58 ) years, and included 27 men (81.82%). There were 30 cases ( 90.91% ) receiving two and more doses of COVID-19 vaccines, and 7 cases ( 21.21% ) with a previous history of SARS-CoV-2 infections. There were 19 cases ( 57.58% ) from African countries, and 7 of 11 cases with Omicron infections were imported from African countries. Conclusion Omicron infection was predominant among individuals positive for SARS-CoV-2 nucleic acid in this centralized isolation site for people entering China from December 18, 2021 to January 12, 2022, with no severe cases detected, and most positive cases were imported from African countries.

Schizophrenia is a serious mental disease. The diagnosis of schizophrenia so far relies heavily on subjective evidence, including self-reported experiences by patients, manifestations described by relatives, and abnormal behaviors assessed by psychiatrists. The diagnosis, monitoring of the disease progression and therapy efficacy assessment are challenging due to the lack of established laboratory biomarkers. Based on the current literature, clinical consensus, guidelines, and expert recommendations, this review highlighted evidence-based potential laboratory biomarkers for the diagnosis of schizophrenia, including genetic biomarkers, neurotransmitters, neurodevelopmental-related proteins, and intestinal flora, and discussed the potential future directions for the application of these biomarkers in this field, aiming to provide an objective basis for the use of these biomarkers in the early and accurate diagnosis, treatment, and prognosis and rehabilitation assessment of schizophrenia.