Objective:Conduct an in-depth investigation into the current status and development needs of infectious disease clinical testing at medical institutions at all levels across the country.Methods:A cross-sectional survey was carried out from January 5, 2023, to June 1, 2023, using a web-based questionnaire distributed to healthcare organizations at all levels nationwide. A total of 3 462 questionnaires were collected, and after quality control and a no-return random sampling process, 2 778 questionnaires were included in the analysis. The research covered basic information about medical institutions, the current emergency response capacity of infectious disease laboratories, the status of the implementation of statutory infectious diseases, the demand for infectious disease testing programs, and the reasons for any lack of implementation. Additionally, the study further analyzed the participation rates of medical institutions at all levels, laboratory accreditation, the presence of infectious disease testing departments, and the testing rates for major infectious diseases.Results:Among the 2 778 questionnaires, the proportions of tertiary, secondary, and primary hospitals were 39.13% (1 087/2 778), 53.20% (1 478/2 778) and 4.46% (124/2 778), respectively. The proportion of specialized infectious disease medical institutions was 8.89% (247/2 778). The proportion of medical institutions with laboratory accreditation was 12.92% (359/2 778). The proportion of clinical laboratory performing infectious disease testing was 98.56% (2 738/2 778). Among the infectious disease pathogens included in the analysis, the implementation rates of testing related to hepatitis B virus, hepatitis C virus, 2019-nCoV, human immunodeficiency virus, and syphilis spirochete were 97.62% (2 712/2 778), 94.85% (2 635/2 778), 93.27% (2 591/2 778), 96.33% (2 676/2 778) and 96.22% (2 673/2 778), respectively. In the research on the demand for clinical infectious disease pathogen testing, the demand for testing of Brucella spp. bacteria and Mycobacterium tuberculosis was 25.50% (90/353) and 8.22% (29/353), respectively. The main factors that did not meet the demand for clinical infectious disease testing were the lack of space in the laboratory to carry out the tests, insufficient manpower, and the absence of a fee schedule, accounting for 44.10% (1 225/2 778), 42.55% (1 182/2 778) and 36.00% (1 000/2 778), respectively.Conclusions:Clinical testing for infectious diseases in our country is mainly carried out in the laboratories of tertiary and secondary medical institutions, which can adequately meet the laboratory testing needs for common infectious diseases. However, there are situations such as limited infectious disease testing projects and insufficient spatial, human, and material resources. Improvements in laboratory construction and methodology can be made according to the actual conditions of each region.

Objective:To explore the protective effect of sesquiterpene lactones of Eupatorium lindleyanum DC.(SLEL)on lipopolysac-charide(LPS)-induced acute lung injury(ALI)in rats using metabolomics.Methods:Forty-eight male SD rats were randomly divided into a blank control group(CG),a model control group(MG),low-,medium-,and high-dose SLEL groups(50,100,and 200 mg/kg),and a positive control group(dexamethasone acetate tablets,5 mg/kg).CG and MG groups were given phosphate-buffered saline.All groups received intragastric administration at a dose volume of 10 mL/kg once a day for 7 consecutive days.One hour after the last ad-ministration,LPS(5 mg/kg)was instilled into the trachea of all groups except the CG group to establish the ALI rat model.Twenty-four hours after model establishment,blood was collected from the abdominal aorta and bronchoalveolar lavage fluid(BALF)was col-lected from the left lung.The total number of inflammatory cells,neutrophils,lymphocytes,and eosinophils in BALF was counted by Wright-Giemsa staining.The levels of interleukin-18(IL-18)and interferon-γ(IFN-γ)in serum and BALF were measured by enzyme-linked immunosorbent assay.The pathological changes of lung tissue were observed using hematoxylin-eosin staining.The ex-pression levels of tight junction protein-1(ZO-1)and occludin in lung tissue were determined by Western blot.The mRNA expression levels of IL-18,IFN-γ,ZO-1,and occludin in the lung were measured by real-time fluorescence quantitative PCR.Non-targeted me-tabolomics analysis of serum and lung tissue was performed using ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry.Results:Compared with the MG group,all SLEL groups had significantly reduced wet/dry weight ratio of lung tissue,lung coefficient,and total number of inflammatory cells,neutrophils,lymphocytes,and eosinophils in BALF.SLEL signifi-cantly decreased the levels of IL-18 and IFN-γ in serum and BALF and the mRNA expression of IL-18 and IFN-γ in lung tissue,and significantly promoted the protein and mRNA expression of ZO-1 and occludin.Under light microscopy,the degree of lung tissue edema,alveolar hemorrhage,and inflammatory cell infiltration were significantly reduced,indicating a certain protective effect on ALI rats.The results of non-targeted metabolomics studies showed that there were 91 and 33 significantly different metabolites in the serum and lung tissue of rats treated with SLEL,respectively.Among them,the main differential metabolites in the serum were sphingosine,L-lactic acid,nicotinic acid,D-nucleotide,and mevalonate-5P,while the main differential metabolites in the lung tissue were tauro-cholic acid.This suggests that SLEL may mainly affect the metabolic pathways of sphingolipids,pyruvate,nicotinic acid,nicotinamide,and tryptophan in the serum and the metabolic pathways of taurine and hypotaurine in the lung tissue to improve ALI.Conclusion:SLEL has a significant protective effect on rats against LPS-induced ALI,and its mechanism of action may be related to the inhibition of inflammatory factors,improvement of lung barrier function,and regulation of related metabolic pathways.

Objective To explore the value of serum matrix metalloproteinase 7 combined with glutamyl transferase and total bile acids in the diagnosis of biliary atresia.Methods 112 children hospitalized in Kunming Children's Hospital with the cholestatic jaundice from July 2023 to September 2024 were selected as the research subjects.According to the surgical exploration,intraoperative cholangiography,liver biopsy and follow-up,the children were divided into the biliary atresia group(BA)(n=52)and non-biliary atresia group(Non-BA)(n=60)respecvely.The age,gender,serum matrix metalloproteinase 7(MMP-7),glutamyl transferase(GGT),alanine aminative aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TB),direct bilirubin(DB),total bile acid(TBA)and aspartate aminotransferase/platelet index(APRI)were compared between the two groups.Statistically significant indicators were included in receiver operating characteristic curve(ROC)analysis,and the area under the ROC curve(AUC)and optimal diagnostic margin(Youden index)were calculated.Results There was no significant difference in age,ALT,AST,DB,TB and APRI levels between the two groups of patients(P>0.05);There was a difference in gender composition ratio between the two groups(P=0.006);MMP-7,GGT,TBA levels in the BA group were significantly higher than those in the Non-BA group,and the difference was statistically significant(P<0.05);the AUC of MMP-7,GGT,and TBA in diagnosing BA were 0.946(95%CI 0.897～0.996),0.857(95%CI 0.789～0.926),0.654(95%CI 0.552～0.755)respectively;When the cut-off value of MMP-7 was 22.37 ng/ml,the sensitivity and specificity for diagnosing BA were 0.923 and 0.933 respectively;When the cut-off value of GGT was 151.5 U/L,the sensitivity and specificity of diagnosing BA were 0.885 and 0.733 respectively;When the cut-off value of TBA was 119.5 μmol/L,the sensitivity and specificity of diagnosing BA were 0.788 and 0.500,respectively.The AUC of MMP-7+GGT and MMP-7+TBA combined to diagnose BA were 0.971(95%CI 0.946～0.997)and 0.943(95%CI 0.889～0.996)respectively.Conclusion Serum MMP-7 has the good diagnostic value as a single indicator for diagnosing BA;MMP-7 combined with GGT is better than a single indicator for diagnosing BA;MMP-7 combined with TBA cannot improve the diagnostic efficiency.

The female reproductive system is essential for sustaining reproductive endocrine homeostasis,however,its vulnerability to various endogenous and exogenous insults,including pathological conditions,pharmacological agents,genetic predispositions,and environmental factors,often results in compromised fertility.The existing protective approaches(including surgical interventions,hormonal replacement therapies,and assisted reproductive techniques)are constrained by several limitations,such as adverse therapeutic effects,technical complexities,and their incapacity to reverse ovarian senescence.Artemisinin and its derivatives(ARTs),characterized by their unique endoperoxide bridge configuration,have exhibited outstanding therapeutic performance across multiple domains including malaria treatment,anticancer therapy,inflammation modulation,and parasitic infection control.Emerging research has identified their novel protective capabilities against various reproductive system pathologies.This comprehensive review systematically elucidates the molecular mechanisms underlying artemisinin-based interventions in reproductive pathologies and evaluates their clinical translation prospects,thereby proposing innovative strategies for the development of next-generation fertility-protective agents with enhanced safety and efficacy profiles.

The female reproductive system is essential for sustaining reproductive endocrine homeostasis,however,its vulnerability to various endogenous and exogenous insults,including pathological conditions,pharmacological agents,genetic predispositions,and environmental factors,often results in compromised fertility.The existing protective approaches(including surgical interventions,hormonal replacement therapies,and assisted reproductive techniques)are constrained by several limitations,such as adverse therapeutic effects,technical complexities,and their incapacity to reverse ovarian senescence.Artemisinin and its derivatives(ARTs),characterized by their unique endoperoxide bridge configuration,have exhibited outstanding therapeutic performance across multiple domains including malaria treatment,anticancer therapy,inflammation modulation,and parasitic infection control.Emerging research has identified their novel protective capabilities against various reproductive system pathologies.This comprehensive review systematically elucidates the molecular mechanisms underlying artemisinin-based interventions in reproductive pathologies and evaluates their clinical translation prospects,thereby proposing innovative strategies for the development of next-generation fertility-protective agents with enhanced safety and efficacy profiles.

Objective:Conduct an in-depth investigation into the current status and development needs of infectious disease clinical testing at medical institutions at all levels across the country.Methods:A cross-sectional survey was carried out from January 5, 2023, to June 1, 2023, using a web-based questionnaire distributed to healthcare organizations at all levels nationwide. A total of 3 462 questionnaires were collected, and after quality control and a no-return random sampling process, 2 778 questionnaires were included in the analysis. The research covered basic information about medical institutions, the current emergency response capacity of infectious disease laboratories, the status of the implementation of statutory infectious diseases, the demand for infectious disease testing programs, and the reasons for any lack of implementation. Additionally, the study further analyzed the participation rates of medical institutions at all levels, laboratory accreditation, the presence of infectious disease testing departments, and the testing rates for major infectious diseases.Results:Among the 2 778 questionnaires, the proportions of tertiary, secondary, and primary hospitals were 39.13% (1 087/2 778), 53.20% (1 478/2 778) and 4.46% (124/2 778), respectively. The proportion of specialized infectious disease medical institutions was 8.89% (247/2 778). The proportion of medical institutions with laboratory accreditation was 12.92% (359/2 778). The proportion of clinical laboratory performing infectious disease testing was 98.56% (2 738/2 778). Among the infectious disease pathogens included in the analysis, the implementation rates of testing related to hepatitis B virus, hepatitis C virus, 2019-nCoV, human immunodeficiency virus, and syphilis spirochete were 97.62% (2 712/2 778), 94.85% (2 635/2 778), 93.27% (2 591/2 778), 96.33% (2 676/2 778) and 96.22% (2 673/2 778), respectively. In the research on the demand for clinical infectious disease pathogen testing, the demand for testing of Brucella spp. bacteria and Mycobacterium tuberculosis was 25.50% (90/353) and 8.22% (29/353), respectively. The main factors that did not meet the demand for clinical infectious disease testing were the lack of space in the laboratory to carry out the tests, insufficient manpower, and the absence of a fee schedule, accounting for 44.10% (1 225/2 778), 42.55% (1 182/2 778) and 36.00% (1 000/2 778), respectively.Conclusions:Clinical testing for infectious diseases in our country is mainly carried out in the laboratories of tertiary and secondary medical institutions, which can adequately meet the laboratory testing needs for common infectious diseases. However, there are situations such as limited infectious disease testing projects and insufficient spatial, human, and material resources. Improvements in laboratory construction and methodology can be made according to the actual conditions of each region.

Objective:To study the clinical features and risk factors of prognosis of neonatal appendicitis.Methods:From January 2014 to December 2022, all infants with neonatal appendicitis and received surgery in our hospital were retrospectively analyzed.Results:A total of 6 cases were enrolled, including 1 boy and 5 girls, with gestational age 36-40 weeks, birth weight 1 990~3 300 g, age of admission 5-11 d and time from illness onset to admission 0.5-4 d. All infants had abdominal distension, combined with vomiting in 4 cases, fever in 3 cases and blood in stool in 1 case. Gastrointestinal perforation was found on preoperative abdominal X-ray in 5 cases. All 6 cases received surgery and confirmed the diagnosis of appendicitis with perforation during the surgery. Appendectomy was performed without mortality. 1 case had Amyand hernia and received high ligation of the hernia sac during operation. 1 case had meningitis and was cured after 3 weeks of antibiotic treatment. 1 case developed adhesive intestinal obstruction 3 months after surgery and underwent intestinal adhesiolysis. One case developed colonic stenosis one month after surgery. The stenotic segment of the colon was resected and primary intestinal anastomosis was performed.Conclusions:Neonatal appendicitis progresses rapidly and is difficult to diagnose. The possibility of appendicitis with perforation should be considered when preoperative abdominal X-ray suggesting pneumoperitoneum. Intraoperatively, it is necessary to pay attention to the relationship between appendiceal perforation and other lesions for comprehensive treatment, and change the surgical approach accordingly.

Efforts have been made to establish various human pluripotent stem cell lines. However, such methods have not yet been duplicated in non-human primate cells. Here, we introduce a multiplexed single-cell sequencing technique to profile the molecular features of monkey pluripotent stem cells in published culture conditions. The results demonstrate suboptimized maintenance of pluripotency and show that the selected signaling pathways for resetting human stem cells can also be interpreted for establishing monkey cell lines. Overall, this work legitimates the translation of novel human cell line culture conditions to monkey cells and provides guidance for exploring chemical cocktails for monkey stem cell line derivation.
Animals ; Haplorhini ; Single-Cell Gene Expression Analysis ; Pluripotent Stem Cells/metabolism* ; Cell Line ; Signal Transduction ; Cell Differentiation ; Transcriptome

Objective:To investigate the influencing factors for synchronous colorectal liver metastasis (synCRLM).Methods:The retrospective case-control study was conducted. The clinicopathological data of 3 172 patients with primary colorectal cancer (CRC) who were admitted to the Affiliated Hospital of Qingdao University from January 2010 to January 2016 were collected. There were 1 946 males and 1 226 females, aged (63±12)years, with a range from 21 to 97 years. Observation indicators: (1) general data analysis; (2) clinicopathological data analysis; (3) analysis of influencing factors for synCRLM. Measurement data with normal distribution were represented as Mean±SD. Count data were represented as absolute numbers. The influencing factors for synCRLM were analyzed after excluding missing data of tumor differentiation degree, tumor diameter, pathological T stage and N stage. Univariate analysis was conducted by chi-square test or Logistic regression model. Multivariate analysis was conducted by Logistic regression model. Results:(1) General data analysis: among the 3 172 patients, cases with age ≤29 years, from 30 to 39 years, from 40 to 49 years, from 50 to 59 years, from 60 to 69 years, from 70 to 79 years, and ≥80 years were 15, 82, 342, 774, 965, 759 and 235, respectively. There were 2 972 patients in Qingdao, 172 cases in Yantai and 28 cases in Weihai. Of the 2 972 patients in Qingdao, there were 422 cases in Shinan District, 658 cases in Shibei District, 457 cases in Huangdao District, 144 cases in Laoshan District, 188 cases in Licang District, 205 cases in Chengyang District, 252 cases in Jimo District, 221 cases in Jiaozhou City, 255 cases in Pingdu City, 170 cases in Laixi City. (2) Clinico-pathological data analysis: among the 3 172 patients, there were 1 639 cases of colon cancer including 972 cases with left colon cancer and 667 cases with right colon cancer, 1 533 cases of rectal cancer. There were 2 981 cases of adenocarcinoma, 165 cases of mucinous adenocarcinoma, 10 cases of signet ring cell carcinoma and 16 cases of other types including carcinoid tumor, squamous carcinoma, tubular adenocarcinoma, etc.There were 162 cases with highly differentiated adenocarcinoma, 5 cases with highly-moderately differentiated adenocarcinoma, 2 338 cases with moderately differentiated adenocarcinoma, 80 cases with moderately-poorly differentiated adeno-carcinoma, 396 cases with poorly differentiated adenocarcinoma and 191 cases missing tumor differentiation data. There were 708 cases with tumor diameter <4 cm, 1 957 cases with tumor diameter ≥4 cm and 507 cases missing tumor diameter data. There were 486 cases in T1 or T2 stage of pathological T stage, 2 169 cases in T3 or T4 stage of pathological T stage and 517 cases missing tumor pathological T staging data. There were 1 563 cases in N0 stage of pathological N staging, 1 062 cases in N1 or N2 stage of pathological N staging and 547 cases missing tumor pathological N staging data. There were 2 895 cases without synCRLM and 277 cases with synCRLM. There were 2 799 cases without diabetes and 373 cases with diabetes. There were 2 931 cases without fatty liver and 241 cases with fatty liver. There were 2 989 cases negative for hepatitis B surface antigen (HBsAg) and 183 cases positive for HBsAg. (3) Analysis of influencing factors for synCRLM. Results of univariate analysis showed that gender, tumor location, tumor differentiation degree, tumor diameter, pathological T stage, fatty liver, HBsAg were related factors for synCRLM in primary colorectal cancer ( χ2=7.400, 7.577, 7.111, 4.513, 12.125, 5.686, 5.919, P<0.05), and neutrophils counts, lymphocytes counts, platelet counts, alanine aminotransferase (ALT), aspartate aminotrans-ferase (AST), total bilirubin, γ-glutamyltransferase (GGT), triacylglycerol (TG), total cholesterol (TC), carcinoembryonic antigen (CEA), and CA19-9 were related factors for synCRLM in primary colorectal cancer ( odds ratio=1.101, 0.807, 1.002, 1.017, 1.023, 1.027, 1.012, 0.686, 1.169, 1.007, 1.004, 95% confidence interval as 1.048-1.156, 0.678-0.960, 1.001-1.004, 1.011-1.024, 1.016-1.031, 1.011-1.044, 1.009-1.015, 0.541-0.869, 1.047-1.306, 1.006-1.008, 1.003-1.004, P<0.05). Results of multivariate analysis showed that cases as male, case with positive HBsAg, AST, GGT, TC, CEA and CA19-9 were independent risk factors for synCRLM in primary colorectal cancer ( odds ratio=1.503, 2.492, 1.018, 1.007, 1.301, 1.005, 1.003, 95% confidence interval as 1.038-2.178, 1.443-4.304, 1.003-1.034, 1.003-1.011, 1.112-1.522, 1.003-1.006, 1.002-1.003, P<0.05), and lymphocytes, ALT and TG were independent protective factors for synCRLM in primary colorectal cancer ( odds ratio=0.777, 0.983, 0.602, 95% confidence interval as 0.608-0.993, 0.966-0.999, 0.421-0.862, P<0.05). Conclusion:Cases as male, case with posotive HBsAg, AST, GGT, TC, CEA and CA19-9 are independent risk factors for synCRLM in primary colorectal cancer, while lymphocytes, ALT and TG are independent protective factors for synCRLM in primary colorectal cancer.

Objective:To construct RNA interference (RNAi) lentiviral expression vector of DEK gene, and to explore its effect on the biological behavior of liver cancer cells.Methods:Double-stranded oligo DNAs were annealed and synthesized according to the interference sequence of DEK gene by RNAi technology. Small interfering RNA expression vector pLKO.1 was cloned after enzymatic digestion. The recombinant lentiviral pLKO.1-sh hDEK was constructed, and then the virus supernatant was collected, packed and infected by 293T cells. Real-time reverse transcription PCR (RT-PCR) and Western blot were used to detect DEK expression in human liver cancer cells Bel-7402, Hu-7, SmMC-7721 and HepG2, and DEK knockdown efficiency in each group of lentivirus-infected cells. Cell proliferation ability, cloning ability, apoptosis and migration ability were detected by cell counting kit-8 (CCK8), flow cytometry and scratch test, respectively. The t-test was used to compare the mean between the two groups, and one-way ANOVA was used to compare the multiple groups.Results:Enzymatic digestion and DNA sequencing results confirmed that the recombinant lentiviral vectors pLKO.1-sh hDEK1 and pLKO.1-sh hDEK3 were successfully constructed. RT-PCR and Western blot results showed that the expression of DEK in human liver cancer cells BEL-7402 and Huh7 cells was higher, and pLKO.1-sh hDEK3 was more effective in inhibiting the DEK gene expression ( P < 0.05). Therefore, pLKO.1-sh hDEK3 was selected to infect BEL-7402 and Huh7 cells for subsequent functional experiments. CCK8 cell proliferation test result showed that the cell proliferation ability of BEL-7402 and Huh7 cells infected with recombinant lentivirus was weakened when compared with blank control and negative control group ( P < 0.05). Apoptosis results showed that the apoptosis rate of knockdown group was higher than that of blank and the negative control group ( P < 0.05). Cell scratch test result showed that the wound healing rate of knockdown group was lower than that of blank control and negative control group ( P < 0.05), and the difference was statistically significant; however, there was no statistically significant difference between blank control and negative control group. Conclusion:Targeting DEK expression in silent liver cancer cells can inhibit the cell proliferation, migration ability, and induce apoptosis, which lays the foundation for further study of the role of DEK gene in liver cancer.