Background: Influenza A virus (IAV) is a negative-sense RNA virus of the Orthomyxoviridae family and is the etiological agent of a highly contagious acute respiratory disease that can lead to acute lung injury. Objective: To elucidate the molecular mechanisms of IAV infection, an integrative research approach combining gene expression profiling, multinetwork analysis, and in vivo experimental validations was employed. Methods: First, a series of network-based analyses were performed, including protein-protein interaction network construction, weighted gene co-expression network analysis, and subsequent gene set enrichment analysis, to identify the major underlying mechanisms of IAV infection. Following gene expression analysis, core targets, both direct and indirect regulators, were screened. An IAV (H1N1) strain A/PR/8/34-induced acute lung injury mouse model was constructed for in vivo validations. Batch one included two groups to evaluate findings from the multi-network analysis: Mock (n = 10; 5 males and 5 females) and IAV (n = 10; 5 males and 5 females). Batch two included three groups to assess the role of toll-like receptor 3 (TLR3) in IAV infection: Mock (n = 6; 3 males and 3 females), IAV (n = 6; 3 males and 3 females), and TLR3 inhibitor (n = 6; 3 males and 3 females). Body weight was measured on days 0, 3, and 5 after infection. On day 5, lung tissues were collected to assess viral load and histopathological changes. Key targets were examined using enzyme-linked immunosorbent assay, Western blotting, and immunofluorescence staining, both in sera and lung tissues. Results: IAV infection was significantly associated with dysregulation of the immune-inflammation system, such as the LTR, nucle-otide-binding oligomerization domain-(NOD) like receptor, retinoic acid-inducible gene I-like receptor, and nuclear factor kappa-B signaling pathways. Gene set enrichment analysis further indicated that the TLR and necroptosis signaling pathways played crucial roles in the progression of IAV infection (TLR signaling pathway normalized enrichment score = 2.3941, P = 1.00 × 10 ⁻¹⁰; necroptosis normalized enrichment score = 1.9421, P = 6.21 × 10 ⁻⁷). Among the core targets, TLR3 and mixed lineage kinase domain-like protein (MLKL) may regulate gene expression at the transcriptional level (all P < 0.05). In vivo validation using an IAV (PR8) infected acute lung injury mouse model demonstrated increased viral load and lung index, alveolar structural damage, and inflammatory cell infiltration. Immunofluorescence staining exhibited large gaps in Lamin B1 staining and breaches in Emerin signals following IAV-PR8 infection. Expression levels of TLR3, p-receptor-interacting serine/threonine-protein kinase 3 (RIPK3)/RIPK3, and p-mixed lineage kinase domain-like protein (MLKL)/MLKL proteins in lung tissues, as well as proinflammatory factors and mediators in sera, were significantly elevated after IAV infection. Moreover, enhanced neutrophil infiltration (myeloperoxidase) and citrullinated histone H3 (a neutrophil extracellular trap-specific marker), both established indicators of neutrophil extracellular trap formation, were observed. Notably, treatment with a TLR3 inhibitor significantly ameliorated IAV-induced acute lung injury by regulating necroptosis-related targets. Conclusion: Our study provides network-based in vivo evidence that TLR3-receptor-interacting serine/threonine-protein kinase 3-MLKL-mediated necroptosis may underlie IAV-induced acute lung injury and could serve as a potential therapeutic target in severe influenza cases.

The diseases and pathological manifestations associated with chongmai (thoroughfare vessel) are the representative in the practical applications of chongmai theory. Elucidating this theory serves as a prerequisite of acupuncture and moxibustion therapy. Based on literature analysis, the diseases and clinical manifestations of chongmai recorded in the time of Huangdi Neijing (The Yellow Emperor's Inner Classic) were analyzed so as to reinterpret the diagnostic process, pathological characteristics, and clinical manifestations of disorders, and evaluate the value of chongmai theory. Chongmai diseases were identified from the palpation initially, known as the "palpation on abdominal pulse". The "qi reversion" of chongmai represents a typical clinical manifestation of chongmai diseases, such as ji (abdominal mass), jia (abdominal hematoma), and shan (hernia), occurring in different sites of the abdomen and in different pathological stages. A part of distribution of chongmai is considered in clinical manifestation, diagnosis and treatment of acupuncture and moxibustion. To emphasize the significance of chongmai in modern acupuncture-moxibustion theory and practice, the diagnostic and therapeutic patterns of the related disorders should be considered in clinical practice. The characteristics of chongmai are reflected in three aspects: qi, blood and abdomen, which are determined by both its inherent properties and the clinical manifestations of related diseases.
Humans ; Acupuncture Therapy ; Moxibustion ; History, Ancient ; Meridians ; Medicine, Chinese Traditional

OBJECTIVE To explore the potential mechanisms and bioactive compounds of licorice against COVID-19 based on a multidimensional interaction of"component-disease-symptom-target".METHODS Firstly,candidate target sets for licorice compo-nents were obtained from the ETCM2.0 database based on the Encyclopedia of Traditional Chinese Medicine,and the disease symp-toms and associated target sets for COVID-19 were derived from the GeneCards and HPO databases.And then a protein interaction network was constructed using the String database(version 12.0).By calculating topological eigenvalues and functional enrichment analysis,the potential mechanisms of action were explored.Combined with Schr?dinger molecular docking virtual screening,candidate pharmacodynamic substances were identified.Fluorescence resonance energy transfer was used for experimental verification.RESULTS Licorice might improve symptoms of COVID-19(fever,chest pain and so on)by regulating the imbalance of"immune-inflammation"network and signal transduction abnormalities during the development and progress of COVID-19,such as coronavirus disease-COVID-19,Toll-like receptor signaling pathway and NOD-like receptor signaling pathway.The components,such as Isos-chaftoside and Hesperidin,were identified to possess high binding affinity with the critical enzymes for viral replication.Isoschaftoside,Hesperidin,Isoliquiritin apioside and Vicenin-2 exhibited varying degrees of inhibition on the enzyme of 3CLpro at different concentra-tions while excluding the interference of the compounds' fluorescence.CONCLUSION Through experimental verification using a multidimensional interaction network of"component-disease-symptom-target",the key pharmacologically active substances of licorice in the fight against COVID-19 are preliminarily identified,and their mechanism of action through overall regulation is elucidated.

OBJECTIVE To explore the potential mechanisms and bioactive compounds of licorice against COVID-19 based on a multidimensional interaction of"component-disease-symptom-target".METHODS Firstly,candidate target sets for licorice compo-nents were obtained from the ETCM2.0 database based on the Encyclopedia of Traditional Chinese Medicine,and the disease symp-toms and associated target sets for COVID-19 were derived from the GeneCards and HPO databases.And then a protein interaction network was constructed using the String database(version 12.0).By calculating topological eigenvalues and functional enrichment analysis,the potential mechanisms of action were explored.Combined with Schr?dinger molecular docking virtual screening,candidate pharmacodynamic substances were identified.Fluorescence resonance energy transfer was used for experimental verification.RESULTS Licorice might improve symptoms of COVID-19(fever,chest pain and so on)by regulating the imbalance of"immune-inflammation"network and signal transduction abnormalities during the development and progress of COVID-19,such as coronavirus disease-COVID-19,Toll-like receptor signaling pathway and NOD-like receptor signaling pathway.The components,such as Isos-chaftoside and Hesperidin,were identified to possess high binding affinity with the critical enzymes for viral replication.Isoschaftoside,Hesperidin,Isoliquiritin apioside and Vicenin-2 exhibited varying degrees of inhibition on the enzyme of 3CLpro at different concentra-tions while excluding the interference of the compounds' fluorescence.CONCLUSION Through experimental verification using a multidimensional interaction network of"component-disease-symptom-target",the key pharmacologically active substances of licorice in the fight against COVID-19 are preliminarily identified,and their mechanism of action through overall regulation is elucidated.

ObjectiveTo systematically explore the underlying mechanisms of Huashi Baidu prescription （HBP） against myocardial injury through a multidimensional network analysis of "transcriptome-putative target-phenotype gene". MethodPutative targets of compounds in HBP were predicted using the Encyclopedia of Traditional Chinese Medicine （ETCM 2.0，http：//www.tcmip.cn/ETCM2/front/） and the Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP v2.0，http：//www.tcmip.cn/TCMIP/index.php/Home/Login/login.html）. Clinical predominant symptom phenotypes for HBP against coronavirus disease 2019 （COVID-19） were collected from CNKI and PubMed databases， while clinical side effects of doxorubicin were gathered from the CTD database. Phenotype-related genes were collected from the HPO database and SoFDA data platform （http：//www.tcmip.cn/Syndrome/front/#/）. An interaction network of "transcriptome-putative target-phenotype gene" was constructed. Key nodes were identified through topological feature calculations， and functional enrichment analysis was conducted to explain the underlying mechanism. Pharmacodynamic evaluation and mechanism verification were performed using a doxorubicin-induced myocardial injury mouse model. Sixty-five SPF-grade C57BL/6J male mice were randomly divided into a control group， a doxorubicin model group， and low-， medium-， and high-dose HBP groups （HBP-L/M/H）， with 13 mice per group. Histopathological severity was assessed using hematoxylin-eosin （HE） and Masson staining. Fibrosis markers were measured by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and protein expression was detected by Western blot. ResultA total of 1 044 putative targets for HBP were obtained from ETCM 2.0 and TCMIP v2.0. From the CTD， HPO， and SoFDA databases， 1 223 potential effect-related targets were identified. Cardiac transcriptome sequencing （RNA-seq） yielded 214 genes related to doxorubicin-induced myocardial injury and 58 genes associated with the effects of HBP， using criteria of P-value<0.05 and FC > 1.5/< 0.667. Analysis of the multidimensional molecular network revealed that the mechanisms of HBP on myocardial injury were mainly related to immune-inflammation imbalance， cellular metabolism， myocardial cell function and fibrosis， angiogenesis， contraction and platelet activation， DNA synthesis， metabolism and repair， as well as cell death and oxidative stress. HBP improved cardiovascular abnormalities such as reduced ejection fraction， myocardial fibrosis， myocarditis， hypertriglyceridemia， and arterial/venous thrombosis. Animal experiments demonstrated that HBP reduced the abnormal high expression of Nod-like receptor heat protein domain Protein 3 （NLRP3）， Caspase-1， apoptosis-associated speck-like protein （ASC）， and Gasdermin D （GSDMD） in myocardial tissue induced by doxorubicin （P<0.05）， and improved the elevated mRNA levels of fibrosis markers transforming growth factor-β₁ （TGF-β₁）， Smad3， and α-smooth muscle actin （α-SMA） （P<0.05）. ConclusionHBP can improve myocardial injury through multiple components， targets， and effects， with the inhibition of the NLRP3 inflammasome activation pathway being a key pharmacodynamic mechanism. This study is expected to provide new insights for the development of targeted therapeutic drugs.

Objective To evaluate the clinical application value of the 5 estimated glomerular filtration rate(eGFR)formulas,inclu-ding MDRD,MDRD Chinese formula[MDRDCHN],CKD-EPICr,CKD-EPICysC and CKD-EPICr-CysC,in assessing glomerular filtration function and diagnosing chronic kidney disease(CKD),and determine the most suitable formula for the local region.Methods A to-tal of 2 610 outpatients and inpatients from The First Affiliated Hospital of Ningbo University were enrolled as the study subjects.Serum creatinine(Cr)and CysC levels were measured,and eGFR was estimated using the 5 formulas.The differences of eGFR values calcu-lated by different formulas were compared.In 412 inpatients,the correlations between endogenous creatinine clearance rate(cCr)and the 5 eGFR results were analyzed,and ROC curves were plotted to compare the diagnostic efficacy of the five eGFR values for CKD.The reference interval for eGFR was established using data from 239 healthy individuals.Results All the eGFR values calculated by the 5 formulas showed skewed distributions and differences of most pairwise comparisons were statistically significant.All the 5 eGFR values were significantly positively correlated with cCr.The CKD-EPICrCysC showed the highest correlation coefficient(r=0.903).The areas under the ROC curves for diagnosing CKD using the 5 eGFR formulas were 0.968,0.969,0.970,0.967 and 0.976,respectively.CKD-EPICr-CysC showed the best diagnostic performance(sensitivity of 97.3％,specificity of 89.8％).The lower limits of the 95％confi-dence interval for healthy individuals were 76,87,82,99,and 93 mL/min/1.73m2 for the different formulas respectively.Conclusion Among the five eGFR formulas,CKD-EPICr-CysC with a reference interval lower limit of 93 mL/min/1.73m2 was demonstrates as the best diagnostic efficiency for assessing glomerular filtration function and diagnosing CKD,and is worth promoting and applying in Ning-bo region.

A 55-year-old male patient was treated with tacrolimus (2.5 mg in the morning and 2 mg at night) for 6 months after lung transplantation to prevent rejection. The blood trough concentration of tacrolimus was stable at 8.0-10.0 μg/L. The patient received antiviral treatment due to corona virus disease 2019 (nirmatrelvir/ritonavir 300 mg/100 mg twice daily orally for a total of 5 days). During the antiviral treatment, the patient continued the anti-rejection treatment. On the second day of antiviral treatment, the patient′s blood trough concentration of tacrolimus increased to >40.0 μg/L, which was considered to be caused by the interaction between nirmatrelvir/ritonavir and tacrolimus. Tacrolimus was withdrawn and antiviral therapy was continued. After discontinuation of tacrolimus for 8 days and nirmatrelvir/ritonavir for 3 days, the blood trough concentration of tacrolimus decreased to 25.7 μg/L. After re-giving tacrolimus at reducing dosage for 3 days, the blood trough concentration of tacrolimus was 8.3 μg/L. After 13 days of resuming administration of tacrolimus at the original dose and frequency, the patient′s blood trough concentration of tacrolimus was 9.2 μg/L. Since then, the blood trough concentration of tacrolimus in the patient was not abnormal again.

A 55-year-old male patient was treated with tacrolimus (2.5 mg in the morning and 2 mg at night) for 6 months after lung transplantation to prevent rejection. The blood trough concentration of tacrolimus was stable at 8.0-10.0 μg/L. The patient received antiviral treatment due to corona virus disease 2019 (nirmatrelvir/ritonavir 300 mg/100 mg twice daily orally for a total of 5 days). During the antiviral treatment, the patient continued the anti-rejection treatment. On the second day of antiviral treatment, the patient′s blood trough concentration of tacrolimus increased to >40.0 μg/L, which was considered to be caused by the interaction between nirmatrelvir/ritonavir and tacrolimus. Tacrolimus was withdrawn and antiviral therapy was continued. After discontinuation of tacrolimus for 8 days and nirmatrelvir/ritonavir for 3 days, the blood trough concentration of tacrolimus decreased to 25.7 μg/L. After re-giving tacrolimus at reducing dosage for 3 days, the blood trough concentration of tacrolimus was 8.3 μg/L. After 13 days of resuming administration of tacrolimus at the original dose and frequency, the patient′s blood trough concentration of tacrolimus was 9.2 μg/L. Since then, the blood trough concentration of tacrolimus in the patient was not abnormal again.

Objective:We employ different regimens of induction therapy in living donor ABO-incompatible kidney transplantation(ABOi-KT) recipients to compare their clinical outcomes during 6 months post-KT.Methods:A retrospective analysis was conducted for the relevant clinical data of 41 ABOi-KT recipients from June 2018 to September 2022.Thirteen recipients on induction therapy of anti-human T lymphocyte porcine immunoglobin(pATG)were enrolled in pATG group; 19 recipients on induction therapy of basiliximab in basiliximab group; 9 recipients on induction therapy of rabbit anti-human thymocyte immunoglobulin(rATG)in rATG group.Differences in age, gender, body mass index(BMI), dialysis modality/duration, sideness of donor kidney, frequency of blood group antibody treatment, dose of rituximab, basic blood group antibody titers of IgG/IgM, and the gender and BMI of recipient's donor were compared for three groups.Immune status was assessed by comparing absolute lymphocyte count before pre-treatment and within 6 months post-KT in recipients under different induction regimens among 3 groups by one-way analysis of variance.Transplant kidney function was assessed by comparing the levels of serum creatinine, estimated glomerular filtration rate(eGFR)and serum urea nitrogen using one-way analysis of variance.The incidence of delayed graft function(DGF), acute rejection(AR)and infection was compared among three groups.Results:Regarding baseline profiles, except for donor age pATG group[(60.23±6.10)years]versus basiliximab group[(51.95±6.97)years]was statistically significant( P=0.002), the differences in the remaining parameters were not statistically significant among three groups(all P>0.05). At Day 1/3/7/10/14 post-KT, absolute lymphocyte counts were(0.17±0.07)×10 9/L, (0.27±0.14)×10 9/L, (0.85±0.40)×10 9/L, (1.05±0.56)×10 9/L and(1.10±0.56)×10 9/L in pATG group and(0.69±0.04)×10 9/L, (0.18±0.21)×10 9/L, (0.57±0.44)×10 9/L, (0.67±0.45)×10 9/L and(0.81±0.46)×10 9/L in rATG group respectively.They were all higher than those in basiliximab group[(0.46±0.18)×10 9/L, (0.67±0.26)×10 9/L, (1.29±0.48)×10 9/L, (1.56±0.49)×10 9/L, (1.75±0.53)×10 9/L]and the differences were statistically significant(all P<0.05). No statistically significant difference existed in absolute lymphocyte count among 3 groups before pre-treatment and after Day 21 post-KT(all P>0.05). At Week 1/2/4/12/24 post-KT, the differences in serum levels of creatinine and urea nitrogen were not statistically significant( P>0.05). At Month 1/3 post-KT, eGFR was(47.24±14.51)and(49.94±14.31)ml·min -1·(1.73 2) -1 in rATG group and they were lower than(67.36±21.60)and(65.00±14.67)ml·min -1·(1.73 2) -1 in basiliximab group with a statistically significant difference( P<0.05). However, at Week 1/2/24 post-KT, no statistically significant difference existed in eGFR among 3 groups( P>0.05). In ATG, basiliximab and rATG groups, DGF(1 case, 1 case, 1 case), AR(2 cases, 2 cases, 1 case)and infection(4 cases, 7 cases, 3 cases)occurred during 6 months post-KT. Conclusions:Through a limited sample of single centers, no statistically significant difference existed in graft function recovery for ABOi-KT recipients on induction therapies of pATG, basiliximab and rATG.And DGF, AR and infections occurred in all three groups.However, there were little inter-group differences.

Objective:To discuss the atypical radiological features of posterior Monteggia fracture and appropriate treatment of the fracture.Methods:A retrospective study was conducted of the 12 patients who had been treated for posterior Monteggia fracture with atypical radiological features at Department of Orthopaedics and Traumatology, Beijing Jishuitan Hospital from July 2019 to December 2020. They were 7 males and 5 females, aged from 18 to 65 years (mean, 46.5 years). Their elbow X-ray and CT scan features included unidentified upper ulnoradial dislocation, presence of triangular or quadrilateral butterfly fracture pieces in front of the fracture end at the level of ulnar coronal process, normal humeroradial joint or forward dislocated radial head, comminuted fracture or anterior edge fracture of the radial head, or backward angulated fracture of the radial neck. The proximal ulnar fractures were fixated with olecranon anatomical locking compression plate or with assistant kirschner wire and tension band fixation; the ulnar coronoid process fractures were fixated with kirschner wire or lag screws or a mini-plate; the radial head fractures were fixated with headless compressing screws or a mini-plate or treated with radial head replacement; the severe injury to the radial collateral ligament was repaired with a suture anchor. Fracture union time, complications and range of elbow motion at the final follow-up were recorded. Elbow function was assessed by Mayo elbow performance score (MEPS).Results:All patients were followed up for 6 to 28 months (mean, 16.4 months). All fractures achieved bony union after 12 to 19 weeks (14.6 weeks). The final follow-ups revealed the following: the range of elbow flexion and extension ranged from 75° to 145°, averaging 100.5°; the range of forearm rotation ranged from 80° to 155°, averaging 132.0°; the MEPS ranged from 50 to 100 points, averaging 86.2 points and yielding 5 excellent, 4 good, 2 fair and 1 poor cases. Elbow stiffness developed in 3 cases.Conclusion:Understanding the atypical radiological features of posterior Monteggia fracture can promote better diagnosis and treatment of the posterior Monteggia fracture in clinic.