Traditional Chinese medicines (TCMs) possess a rich historical background, unique theoretical framework, remarkable therapeutic efficacy, and abundant resources. However, the modernization and internationalization of TCMs have faced significant obstacles due to their diverse ingredients and unknown mechanisms. To gain deeper insights into the phytochemicals and ensure the quality control of TCMs, there is an urgent need to enhance analytical techniques. Currently, two-dimensional (2D) chromatography, which incorporates two independent separation mechanisms, demonstrates superior separation capabilities compared to the traditional one-dimensional (1D) separation system when analyzing TCMs samples. Over the past decade, new techniques have been continuously developed to gain actionable insights from complex samples. This review presents the recent advancements in the application of multidimensional chromatography for the quality evaluation of TCMs, encompassing 2D-gas chromatography (GC), 2D-liquid chromatography (LC), as well as emerging three-dimensional (3D)-GC, 3D-LC, and their associated data-processing approaches. These studies highlight the promising potential of multidimensional chromatographic separation for future phytochemical analysis. Nevertheless, the increased separation capability has resulted in higher-order data sets and greater demands for data-processing tools. Considering that multidimensional chromatography is still a relatively nascent research field, further hardware enhancements and the implementation of chemometric methods are necessary to foster its robust development.

Objective:To explore the application effect of intelligent referral mode in examinees with major positive results in physical examinations in a health medical center.Methods:A cross-sectional study was conducted from January 2022 to December 2023 with subjects receiving physical examinations. Those subjects with major positive results from January 2022 to December 2022 in the Health Medical Center of the Second Affiliated Hospital of Chongqing Medical University were selected as the routine referral group, and those with major positive results from January 2023 to December 2023 were selected as the intelligent referral group. The routine referral group received routine health education from the chief inspector in the health medical center, suggesting them to get outpatient consultation. On the basis of ordinary referral, the intelligent referral group received health education from a referral team composed of nurses and specialists based on internet technology to digitally integrate and share patients′ medical records, examination results, imaging data and other medical information. The subects in the intelligent referral group were provided with a green channel for outpatient consultation and medical treatment immediately. The visiting rate, hospitalization rate, average visiting time, and overall satisfaction of the subjects after being notified of abnormal results were compared between the two groups. According to the positive results, the subjects were divided into four subgroups: ultrasound examination group, radiology group, clinical laboratory group and blood pressure group.Results:After the notification, there were significant differences in hospital visit rate (84.8% vs 67.2%, χ 2=168.4), hospitalization rate (48.8% vs 36.5%, χ 2=17.80), average visit time (116 min vs 301 min, Z=-15.82), and overall satisfaction score (9.70±0.62 vs 8.29±1.26, t=-33.47) between intelligent referral group and general referral group (all P<0.01). The classification statistics of major abnormal results showed that the consultation rates in subjects with ultrasound, radiology, clinical laboratory and blood pressure abnormities in the intelligent referral group were all higher than those in the general referral group (87.9% vs 70.4%, 89.9% vs 70.6%, 80.6% vs 60.2%, 57.2% vs 41.3%, respectively; χ 2=41.91, 39.37, 19.37, 6.20, all P<0.05); the hospitalization rates in subjects with ultrasound, radiology and blood pressure abnormities in the intelligent referral group were all higher than those in the general referral group (66.8% vs 64%, 55.7% vs 42.2%, 18.7% vs 11.2%, respectively; χ 2=16.86, 11.91, 8.68, all P<0.05); the mean consultation times in subjects with ultrasound, radiology and clinical laboratory abnormities in the intelligent referral group were all significantly shorter than those in the general referral group (96 min vs 308 min, 110 min vs 300 min, 122 min vs 286 min, Z=-11.38, -9.27, -7.63, all P<0.01); the overall satisfaction scores in subjects with ultrasound examination, radiology, clinical laboratory and blood pressure abnormities in the intelligent referral group were all higher than those in the general referral group (9.69±0.60 vs 8.36±1.21, 9.09±0.62 vs 8.26±1.27, 9.74±0.69 vs 8.25±1.31; 9.68±0.59 vs 8.34±1.35, respectively; t=-18.47, -18.52, -14.42, -11.77, all P<0.01). Conclusions:The application of intelligent referral mode can improve the visiting rate and hospitalization rate of examinees with major positive results of physical examination, shorten their visiting time, and thus improve their satisfaction.

Objective To investigate the effect of silk fibroin hydrogel loaded with developmental endothelial locus-1(Del-1)nanoparticles on the healing of chronic skin wounds in mice.Methods The back skin of BALB/c mice(6-8 weeks old)was pressed with a magnet for 12 h and then relaxed for 12 h,for 4 consecutive days to establish a chronic pressure ulcer wound.After infliction,the mice were randomly divided into 3 groups(n=8),and the skin wounds were treated with PBS,silk fibroin hydrogel or Del-1 nanoparticles/silk fibroin hydrogel.The wound healing was recorded with camera to calculate the wound healing rate.In 9 d after treatment,HE and Masson staining were used to observe the wound healing,and immunofluorescence staining for CD 14 and TNF-α was used to compare the appearance frequency of skin macrophages and the expression of inflammatory factors.After Tert-butyl peroxide(TBHP)was used to stimulate mouse macrophage RAW 264.7 cells and mouse vascular endothelial C166 cells,C166 cells were transfected with lentival vector to overexpress Del-1.Crystal violet staining was used to observe the migration of macrophages.RT-qPCR was used to detect the expression of inflammatory factor IL-6.Results The wound healing was significantly faster in the Del-1 nanoparticles/silk fibroin hydrogel group than the silk fibroin hydrogel group and the PBS group(P＜0.01).The expression levels of TNF-α and CD14 in the wound surface were lower(P＜0.01),but collagen deposition and tissue repair were better in the Del-1 nanoparticles/silk fibroin hydrogel group than the silk fibroin hydrogel group and the PBS group(P＜0.01).In vitro experiments,macrophages migrated to endothelial cells stimulated by TBHP,but the migration rate of macrophages was significantly lower in the Del-1 overexpression group(P＜0.01).RT-qPCR confirmed that Del-1 inhibited the transcription of IL-6(P＜0.01).Conclusion Del-1 nanoparticles/silk fibroin hydrogel can significantly accelerate the healing of skin wounds,and its mechanism may be through promoting the regression of inflammation and tissue repair.

Objective:To evaluate effect of diammonium glycyrrhizinate(DG)on lipopolysaccharide(LPS)-induced TLR4/NF-κB inflammatory signaling pathway in BV2 microglial cells and to explore its potential regulatory mechanisms on ameliorating neu-roinflammation.Methods:BV2 microglia injury model was induced by LPS and treated with 20 and 60 μmol/L DG.MTS was used to determine vitality of cells.NO level secreted by cells was detected by Griess.Inflammatory factors TNF-α,IL-6,and IL-1β levels in cell supernatant were measured by ELISA.TNF-α,IL-6,IL-1β,TLR4 and MyD88 mRNA levels were detected by RT-qPCR.Western blot was used to determine expressions of TLR4,MyD88,NF-κB,p-NF-κB,IκB-α and p-IκB-α proteins of cells.Results:Compared with control group,LPS-treated BV2 microglia had significantly lower viability(P<0.05),significantly higher NO secretion(P<0.05),significantly up-regulation levels of inflammatory factors TNF-α,IL-6 and IL-1β(P<0.05),and significantly higher mRNA levels of TNF-α,IL-6,IL-1β,TLR4 and MyD88(P<0.05),TLR4,MyD88,NF-κB,p-NF-κB,IκB-α,and p-IκB-α protein expressions were significantly increased(P<0.05).Compared with LPS group,BV2 microglia intervened with different concentrations of DG had significantly higher viability(P<0.05),significantly lower NO secretion(P<0.05),significantly lower TNF-α,IL-6 and IL-1β inflammatory factors levels(P<0.05),significantly lower mRNA levels of TNF-α,IL-6,IL-1β,TLR4 and MyD88(P<0.05),and TLR4,MyD88,p-NF-κB and p-IκB-α protein expressions were significantly reduced(P<0.05).Conclusion:DG can inhibit LPS-induced neuroinflammatory responses in microglia,whose underlying mechanism is suppression of TLR4/NF-κB signaling pathway.

Objective:To investigate the positive rate and isolate Bordetella pertussis in children with suspected whooping cough and their close family members in Zhejiang Province, and further explore the susceptibility and resistant mechanism of Bordetella pertussis to antibiotics. Methods:A total of 273 nasopharynx swabs specimens from children with suspected whooping cough in Hangzhou Children′s Hospital from May 2022 to October 2022 were collected. The strains were isolated and cultured using charcoal select agar plate. Pertussis target genes were detected by RT-PCR. E-test method was used to detect the sensitivity of Bordetella pertussis strains to different antibiotics. The mechanism of resistance of Bordetella pertussis to macrolides was analyzed by whole genome sequencing. The phylogenetic analysis of isolated strains was based on core genome multilocus sequence typing(cgMLST). Results:Among 273 clinical samples of children with suspected pertussis and their close family members, 168 samples were positive by fluorescence quantitative PCR, accounting for 61.54%, and 30 pertussis strains were successfully isolated with a positive rate of 10.98%. In addition, among the 143 samples of close family members, 54.55% (78/143) samples were positive by RT-PCR and 9.79% (14/143) samples were positive by culture, suggesting that the close family member are important in family transmission of pertussis. Besides, most of the positive samples were from mothers. The results of E-test showed that 96.67%(29/30) strains showed high resistance to azithromycin with MIC value>256 mg/L, and the resistant mechanism of azithromycin was A2047G mutation in 23S rRNA. The phylogenetic analysis based on the cgMSLT showed that the isolated strains were clustered into two new different clades.Conclusions:The positive rate of Bordetella pertussis in close family members is at a high level and the mother may be the main source of infection, which is of great significance for monitoring and laboratory detection of suspected children′s family members. Bordetella pertussis shows high resistance to macrolides in Zhejiang Province, so monitoring of the antimicrobial resistance should be further strengthened to provide theoretical basis for clinical treatment and drug guidance.

In growing children, growth plate cartilage has limited self-repair ability upon fracture injury always leading to limb growth arrest. Interestingly, one type of fracture injuries within the growth plate achieve amazing self-healing, however, the mechanism is unclear. Using this type of fracture mouse model, we discovered the activation of Hedgehog (Hh) signaling in the injured growth plate, which could activate chondrocytes in growth plate and promote cartilage repair. Primary cilia are the central transduction mediator of Hh signaling. Notably, ciliary Hh-Smo-Gli signaling pathways were enriched in the growth plate during development. Moreover, chondrocytes in resting and proliferating zone were dynamically ciliated during growth plate repair. Furthermore, conditional deletion of the ciliary core gene Ift140 in cartilage disrupted cilia-mediated Hh signaling in growth plate. More importantly, activating ciliary Hh signaling by Smoothened agonist (SAG) significantly accelerated growth plate repair after injury. In sum, primary cilia mediate Hh signaling induced the activation of stem/progenitor chondrocytes and growth plate repair after fracture injury.
Mice ; Animals ; Hedgehog Proteins/genetics* ; Receptors, G-Protein-Coupled/metabolism* ; Cilia/metabolism* ; Cartilage/metabolism* ; Regeneration

Objective:To analyze and discuss the application value and teaching effect of simulate patients (SP)-assisted problem-based learning (PBL) combined with team-based learning (TBL) mode in pediatric first aid teaching, in order to provide a new reference for the reform of standardized residency training and teaching mode of pediatrics in the new era in China.Methods:A total of 103 residents who participated in the standardized training in the Department of Pediatrics of Hunan Children's Hospital from January 2018 to December 2020 were selected as the research objects, and they were divided into the control group (50 cases) and the experimental group (53 cases). The control group adopted the previous teaching mode of residents, while the experimental group adopted the SP-assisted PBL combined with TBL mode. In this study, SPSS 22.0 software was used for t test and chi-square test. Results:The examination scores of the experimental group in the theoretical knowledge of pediatric first aid, clinical thinking and skills, and application of pediatric first aid skills were higher than those in the control group [(86.98±3.42), (85.69±5.13), (89.62±4.75)] vs. [(77.23±4.16), (81.16±3.96), (76.54±5.78)], with statistically significant differences between the two groups ( P<0.05). The questionnaire survey showed that the clinical thinking and pediatric emergency skills, doctor-patient communication ability, teamwork ability and other aspects of the experimental group were significantly better than those of the control group ( P<0.05). Conclusion:The SP-assisted PBL combined with TBL mode can produce positive effects in the standardized residency training of pediatrics, which will help residents better master pediatric first aid skills, improve their clinical comprehensive capabilities, and be more conducive to improving the quality of pediatric residency training, which is worth promoting in the new era.

Objective:To explore the genetic characteristics and evolution of hantavirus carried by rodents in port area of Ningde in Fujian province in the summer of 2020.Methods:Rodents were captured in the port area of Ningde, the RNA was extracted from rodent lung tissues and detected by using specific kit. The positive samples were used for whole-genome sequencing of the virus. Bioinformatics software was used for the analysis on the similarity and genetic variation of the sequences.Results:A total of 112 rodents were captured, including 5 Rattus norvegicus and 2 Rattus flavipectus, the positive rate of hantavirus was 6.25% (7/112). By virus gene sequencing, two hantavirus complete genome sequences were obtained (named as FJ35 and FJ36, GenBank accession numbers: MW449188-MW449193). The genetic analysis results showed that the hantavirus detected in positive samples were SEOV and shared 99% nucleotide similarity with hantavirus strains LZSF21 and JX20140581 isolated from Shandong province. Phylogenetic analysis using the maximum likelihood method showed that the hantavirus detected in positive samples belonged to S3 subtype, sharing the same subtype with hantavirus strains Z37 from Zhejiang province, LZSF21 from Shandong province, and zy27 and Gongzhuling 415 from northeastern China. Compared with FJ372, the amino acid variation of N259S was observed at sites 251-264 of nucleoprotein, which might be related to antigenicity. Another variation of Q81R was observed in glycoprotein compared with SEOV 80-39 segment of coded amino acid of international reference strain, which might also cause the change in antigenicity. Conclusion:The high positive rate of hantavirus in rodents in the port area of Ningde- would increase the risk of natural human infection and epidemic in local area. The hantavirus positive rodents in this focus might be from an endemic area in Shandong. It is necessary to strengthen the imported rodent control in the port area of Ningde. The virus detected in 2 positive samples belonged to SEOV subtype Ⅲ and shared high homologies of nucleotides and amino acid sequences with the hantavirus strains in surrounding area. However, some slight variations occurred in glycoprotein and nucleoprotein amino acid sequences, which might cause changes in its antigeniity.

Objective: By investigating the genotype and evolutionary variation of hantavirus (HV) in Tiantai county, a national surveillance site for hemorrhagic fever with renal syndrome (HFRS) was set in Zhejiang province, from 2011 to 2018, to reveal the molecular epidemiological characteristics of hantavirus (HV) in Tiantai. Methods: Total RNA was extracted from ultrasound treated HV antigen- positive rat lung samples in Tiantai from 2011 to 2018. After cDNA was prepared, nested PCR was used to amplify partial sequence of M fragments by using specific primers of HV. The sequences of HV in Tiantai from 2011 to 2018 were compared with other known HV sequences in order to identify the genotype and analyze the evolution and variation of the virus. Results: In 67 HV antigen-positive lung specimens, 31 were positive in nested PCR amplification with type-specific primers, including 30 Hantaan virus (HTNV) positive samples, 1 Seoul virus (SEOV) positive sample, and all the 31 samples were from Apodemus agrarius. The phylogenetic tree based on partial M segment was divided into monophyletic group, 30 strains were distributed in HTNV group and 1 was in SEOV group. The HTNV strain Tiantai T2018-130 was independently in one branch, sharing 84.8%-87.9% homology with other strains both at home and abroad, including 29 strains in HTNV group in Tiantai. The other 29 HTNV strains in Tiantai showed closer relationship. The SEOV strain T2016-31 from Apodemus agrarius showed closer relationship with previous strains of SEOV, Tiantai ZT71, ZT10 and Z37 strains of Wenzhou, Zhejiang province. Conclusions HTNV, the main genotype of HV in Tiantai of Zhejiang province, showed obvious geographic clustering, but the strain T2018-130 was distinct from the others in Tiantai. Meanwhile, by sequence analysis, we confirmed that The SEOV strain T2016-31 existed in in Apodemus agrarius, indicating there was a phenomenon of "spillover" between virus and host in SEOV evolution.

Objective: To learn the population,virus status and viral types of hantavirus（HV）hosts in Tiantai County of Zhejiang Province from 2011 to 2018,and to provide evidence for hemorrhagic fever with renal syndrome（HFRS）control. Methods: Rodents in Tiantai County were captured by night trapping method. After the species and age of rodents were identified,the composition of rodent species,dominant species and density of rodents were analyzed. The lungs and blood of rodents were sampled to detect the antigen and antibody of HV by immunofluorescence method. The HV antigen-positive lung samples were detected by RT-PCR with specific primers of HV S fragment,then HV was isolated and identified by inoculating Vero-E6 cells. Results: The average rodent density in Tiantai County from 2011 to 2018 was 4.44%. The rodent density in the field and residential areas were 4.94% and 2.23%,respectively. Ten species of rodents were identified,with Apodemus agrarius dominant in the field and Rattus norvegicus in the residential areas. Sixty-seven lung samples were HV antigen positive（4.13%）,one from Rattus norvegicus and the other sixty-six from Apodemus agrarius. Seventy-nine blood samples were HV antibody positive（4.86%）,all from Apodemus agrarius. Thirty-four HV antigen-positive lung samples were positive（50.75%）after RT-PCR amplification. Twenty-two strains of virus were isolated and all of them were from Apodemus agrarius,including twenty-one strains of Hantaan type（HTN）and one strain of Seoul type（SEO）. Conclusion In Tiantai County,Apodemus agrarius is the main source of HFRS infection;the main epidemic type of HV is HTN and SEO is first found in Apodemus agrarius.