ObjectiveTo investigate the optimal ratio of goat horn replacing antelope horn in Fufang Lingjiao Jiangya pills and the blood pressure-lowering mechanism of this medicine. MethodsThe blood pressure-lowering efficacy of Fufang Lingjiao Jiangya pills with varying proportions of goat horn replacing antelope horn was evaluated on spontaneous hypertensive rats （SHR）. In this experiment， 50 SHR rats were randomly grouped as follows： model （n=8）， captopril （0.01 g·kg^-1） （n=6）， low-dose blank Fufang Lingjiao Jiangya pills （0.342 g·kg^-1） （n=6）， high-dose blank Fufang Lingjiao Jiangya pills （0.684 g·kg^-1） （n=6）， low-dose antelope horn-containing Fufang Lingjiao Jiangya pills （0.378 g·kg^-1） （n=6）， high-dose antelope horn-containing Fufang Lingjiao Jiangya pills （0.756 g·kg^-1） （n=6）， low-dose goat horn-containing Fufang Lingjiao Jiangya pills （0.378 g·kg^-1） （n=6）， and high-dose goat horn-containing Fufang Lingjiao Jiangya pills （0.756 g·kg^-1） （n=6）. Additionally， 8 WKY rats were used as the normal group. Drugs were administered by gavage for 4 weeks while an equal volume of distilled water was administered for the normal and model groups. Blood pressure was measured before administration， 3 h post administration， and biweekly thereafter. In the experiment for Fufang Lingjiao Jiangya pills with goat horn replacing antelope horn in different proportions， 48 SHR rats were randomly grouped as follows： model， blank Fufang Lingjiao Jiangya pills （0.684 g·kg^-1）， antelope horn-containing Fufang Lingjiao Jiangya pills （0.756 g·kg^-1）， 2× goat horn-containing Fufang Lingjiao Jiangya pills （0.824 g·kg^-1）， 4× goat horn Fufang Lingjiao Jiangya pills （0.969 g·kg^-1）， and 6× goat horn Fufang Lingjiao Jiangya pills （1.112 g·kg^-1）. The normal group included 8 WKY rats， and the normal group and model group received an equal volume of distilled water. The treatment lasted for 2 weeks， and blood pressure was recorded at various time points （pre-administration， 3 h post administration， and on days 4， 7， 10， and 14 of administration）. Serum levels of angiotensin-converting enzyme （ACE）， angiotensin Ⅱ（Ang Ⅱ）， renin， and interleukin-6 （IL-6） were measured by enzyme-linked immunosorbent assay. Histopathological changes in the heart， kidney， and thoracic aorta were observed by hematoxylin-eosin staining. The protein levels of ACE2， angiotensin Ⅱ type 1 receptor （AT1R）， and angiotensinogen （AGT） in the kidney tissue were determined by Western blot， while the expression of nuclear factor （NF）-κB p65 and Toll-like receptor 4 （TLR4） in the thoracic aorta tissue was assessed by immunohistochemistry. ResultsCompared with the model group， all treatment groups showed lowered blood pressure （P<0.05， P<0.01）， and the 6× goat horn-containing Fufang Lingjiao Jiangya pills group showed consistent blood pressure-lowering effect with the antelope horn-containing Fufang Lingjiao Jiangya pills group. Compared with the normal group， the model group showed elevated serum levels of ACE， Ang Ⅱ， renin， and IL-6， while the elevations were declined in the Fufang Lingjiao Jiangya pills groups （P<0.05， P<0.01）. Pathological changes in the heart， kidney， and thoracic aorta were alleviated in all the treatment groups， with the 6× goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups exhibited the best effect. Western blot and immunohistochemistry results showed that all the treatment groups exhibited down-regulated protein levels of AT1R， AGT， NF-κB p65， and TLR4 and up-regulated protein levels of ACE2 （P<0.05， P<0.01） compared with model group， with the 6×goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups showcasing the best effect. ConclusionReplacing antelope horn with 6×goat horn in Fufang Lingjiao Jiangya pills can achieve consistent blood pressure-lowering effect with the original prescription. The prescription may exert the effect by inhibiting the renin-angiotensin-aldosterone system （RAAS） and TLR4/NF-κB signaling pathways.

Rheumatoid arthritis (RA) is an autoimmune disease with a complex etiology. Monocyte-derived macrophages (MDMs) infiltration are associated with RA severity. We have reported the deletion of G-protein-coupled receptor kinase 2 (GRK2) reprograms macrophages toward an anti-inflammatory phenotype by recovering G-protein-coupled receptor signaling. However, as more GRK2-interacting proteins were discovered, the GRK2 interactome mechanisms in RA have been understudied. Thus, in the collagen-induced arthritis mouse model, we performed genetic GRK2 deletion using GRK2^f/fLyz2-Cre^+/- mice. Synovial inflammation and M1 polarization were improved in GRK2^f/fLyz2-Cre^+/- mice. Supporting experiments with RNA-seq and dual-luciferase reporter assays identified peroxisome proliferator-activated receptor γ (PPARγ) as a new GRK2-interacting protein. We further confirmed that fms-related tyrosine kinase 1 (Flt-1), which promoted macrophage migration to induce angiogenesis, was inhibited by GRK2-PPARγ signaling. Mechanistically, excess GRK2 membrane recruitment in CIA MDMs reduced the activation of PPARγ ligand-binding domain and enhanced Flt-1 transcription. Furthermore, the treatment of mice with GRK2 activity inhibitor resulted in significantly diminished CIA pathology, Flt-1⁺ macrophages induced-synovial inflammation, and angiogenesis. Altogether, we anticipate to facilitate the elucidation of previously unappreciated details of GRK2-specific intracellular signaling. Targeting GRK2 activity is a viable strategy to inhibit MDMs infiltration, affording a distinct way to control joint inflammation and angiogenesis of RA.

AIM:To identify transcriptional differences between the ocular surface ectoderm(OSE)and surface ectoderm(SE)using RNA-seq, and elucidate the OSE transcriptome landscape and the regulatory networks involved in its development.METHODS:OSE and SE cells were differentiated from human embryonic stem(hES)cells. Differentially expressed genes(DEGs)between OSE and SE were analyzed using RNA-seq. Based on the DEGs, we performed gene ontology(GO)analysis, Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis, and protein-protein interaction(PPI)network analysis. Transcription factors(TFs)and hub genes were screened. Subsequently, TF-gene and TF-miRNA regulatory networks were constructed using the NetworkAnalyst platform.RESULTS:A total of 4 182 DEGs were detected between OSE and SE cells, with 2 771 up-regulated and 1 411 down-regulated genes in OSE cells. GO-BP analysis revealed that up-regulated genes in OSE were enriched in the regulation of ion transmembrane transport, axon development, and modulation of chemical synaptic transmission. Down-regulated genes were primarily involved in nuclear division, chromosome segregation, and regulation of cell cycle phase transition. KEGG analysis indicated that up-regulated genes in OSE cells were enriched in signaling pathways such as cocaine addiction, axon guidance, and amphetamine addiction, while down-regulated genes were enriched in proteoglycans in cancer, ECM-receptor interaction, protein digestion and absorption, and cytokine-cytokine receptor interaction. Additionally, compared with SE, 204 TFs(including FOS, EGR1, POU5F1, SOX2, and PAX6)were up-regulated, and 80 TFs(including HAND2, HOXB6, HOXB5, HOXA5, and HOXB8)were down-regulated in OSE cells. Furthermore, we identified 6 up-regulated and 9 down-regulated hub genes in OSE cells, and constructed TF-gene and TF-miRNA regulatory networks based on these hub genes.CONCLUSIONS:The transcriptome characteristics of OSE and SE cells were elucidated through RNA-seq analysis. These findings may provide a novel insight for studies on the development and in vitro directed induction of OSE and corneal epithelial cells.

Objective To generate minichromosome maintenance protein 2(MCM2)gene knockout cervi-cal cancer HeLa cell lines using inducible CRISPR/Cas9 technology,and to explore the effect of MCM2 on DNA replication and replication stress.Methods The inducible CRISPR/Cas9 system,TLCV2,was used to construct MCM2 knockout HeLa cell lines.And the cell lines were divided into control group(Control),knockout group 1(KO1),and knockout group 2(KO2).Western blot,Edu incorporation experiment,real-time quantitative PCR(qPCR),immunofluorescence and MTT assay were used to analyze the effects of MCM2 knockout on DNA replica-tion and replication stress induced by hydroxyurea.Results The CRISPR/Cas9 system successfully knocked out the MCM2 gene after induction,and MCM2 knockout affected the stability of MCM2-7 complex.Compared with the control cells,MCM2 knockout cells had a dramatic decrease in the capacity of DNA replication,and the mRNA levels of Cyclin A1,Cyclin E1 and CDK4.Under DNA replication stress,MCM2 knockout cells decreased cell viability,DNA damage repair capacity,and increased genomic instability compared with control cells.Conclusion Knockout of MCM2 gene reduces the DNA replication capacity of HeLa cells under normal conditions and cell viability under replication stress.This study successfully generates MCM2 gene inducible knockout HeLa cell lines,laying the foundation for further research on the role and biological function of MCM2 gene in the occurrence and progression of cervical cancer.

Foot bath technique is booming in daily health care services. In order to develop a more standardized and complete technical operation process, the working group recorded the whole process of document formulation while writing the Technical Operation Specification for TCM Health Care Services (Non-Medical) Foot Bath (2024 edition). Work profile (including task background, task source, drafting unit, participant grouping and responsibilities), main technical content (including the basis and principles of preparation, key technical content of technical specifications), main preparation process (formation of working groups, registration and plan writing, selection and determination of clinical issues, literature research, drafting of the first draft, consensus on the main content of technical specifications, soliciting opinions, testing applications and external review), the relationship of current mandatory national standards or policies and regulations, the treatment process and basis of major differences, publicity and implementation and post-effect evaluation, the proposal to abolish the current relevant guidelines and the corresponding annexes were under detailed and in-depth description, which can assist the relevant practitioners of non-medical institutions to better understand and apply this technical specification.

Objective:To construct a nomogram model for differentiating gastric schwannoma (GS) from gastric stromal tumor (GST) (diameters 2 to 5 cm) based on CT imaging features before surgery.Methods:The clinical and imaging data of 49 patients with GS and 240 patients with GST in the Affiliated Hospital of Jining Medical University from July 2009 to April 2023 and Guangdong Provincial People′s Hospital from June 2017 to September 2022 were analyzed retrospectively. The independent factors for differentiating GS from GST were obtained by multivariate Logistic regression analysis. The nomogram model was constructed by R4.3.1 software. The efficacy of the nomogram model for differentiating GS from GST was evaluated by the receiver operating characteristics (ROC) curve, and calibration curve and decision curve analysis were used to evaluate the predictive efficacy and clinical application value of the nomogram model.Results:There were no statistical differences in the clinical symptom rate, calcification rate, ulcer rate, tumor vessel rate, ratio of long diameter to short diameter and CT value difference during the arterial and nonenhanced phases (CTV A-N) between GS patients and GST patients ( P>0.05). The proportion of female, incidence of lesions located in central or lower part of stomach, extraluminal or mixed growth rate, tumor-associated lymph node rate, strong enhancement rate, CT value difference during the portal and nonenhanced phases (CTV P-N), CT value difference during the delayed and nonenhanced phases (CTV D-N), CT value difference during the portal and arterial phases (CTV P-A) and CT value difference during the delayed and portal phases (CTV D-P) in GS patients were significantly higher than those in GST patients: 75.51% (37/49) vs. 58.33% (140/240), 85.71% (42/49) vs. 54.17% (130/240), 75.51% (37/49) vs. 45.00% (108/240), 44.90% (22/49) vs. 5.42% (13/240), 51.02% (25/49) vs. 27.08% (65/240), 32.0 (26.0, 43.5) HU vs. 29.0 (22.0, 37.7) HU, (44.59 ± 13.46) HU vs. (32.94 ± 12.47) HU, 20.0 (11.5, 25.0) HU vs. 10.0 (5.0, 17.0) HU and 9.0 (6.0, 12.0) HU vs. 4.0 (-2.7, 7.0) HU, the age, irregular shape rate, cystic degeneration rate and heterogeneous enhancement rate were significantly lower than those in GST patients: (58.12 ± 12.59) years old vs. (62.05 ± 11.22) years old, 16.33% (8/49) vs. 38.33% (92/240), 18.37% (9/49) vs. 51.25% (123/240) and 34.69% (17/49) vs. 56.25% (135/240), and there were statistical differences ( P<0.05 or<0.01). Multivariate Logistic regression analysis result showed that location, cystic degeneration, tumor-associated lymph node, CTV P-A and CTV D-P were the independent factors for differentiating GS from GST ( OR= 3.599, 0.201, 19.031, 1.124 and 1.160; 95% CI 1.184 to 10.938, 0.070 to 0.578, 6.159 to 58.809, 1.066 to 1.185 and 1.094 to 1.231; P<0.05 or<0.01). The nomogram model for differentiating GS from GST was constructed based on location, cystic degeneration, tumor-associated lymph node, CTV P-A and CTV D-P. The area under curve of the nomogram model for differentiating GS from GST was 0.924 (95% CI 0.887 to 0.951). The calibration curve analysis result showed that there was a good agreement between the predicted GS curve and the actual GS curve (the mean absolute error was 0.033). The result of the Hosmer-Lemeshow goodness-of-fit test indicated that the calibration of the nomogram model was appropriate ( χ2 = 2.52, P = 0.961). The clinical decision curve analysis result showed that when the threshold for the nomogram model for differentiating the two tumors was>0.03, the nomogram yielded more net benefits than the "all patients treated as GS" or "all patients treated as GST" scenarios. Conclusions:The nomogram model based on CT imaging features can be used to differentiate GS from GST before surgery.

Objective To assess the risk factors for systemic inflammatory response syndrome(SIRS)induced by flexible ureteroscopic lithotripsy(FURL)and compare the diagnostic value of bacterial culture for stone,midstream urine culture and pelvic urine culture in predicting postoperative infection.Methods Patients who underwent FURL for upper urinary tract stones from November 2021 to May 2023 were retrospectively enrolled.The patients were divided into non-SIRS and SIRS groups.We collected midstream urine one day before surgery and stone and pelvic urine samples during surgery.Multivariate lo-gistic regression analysis was used to identify the risk factors for SIRS induced by FURL.Results Totally,326 patients,11.66％(38/326)developed SIRS,and 4.29％(14/326)had severe sepsis.The percentages of positive bacterial cultures for mid-stream urine,pelvic urine and stone cultures were 16.87％(55/326),10.43％(34/326)and 15.95％(52/326),respectively.In the multivariate logistic regression analysis,stone size(P=0.010),operation time(P=0.030),and stone culture(P＜0.01)were identified as independent risk factors for SIRS induced by FURL.Positive bacterial culture for stone was an independent risk factor for postoperative severe sepsis(P=0.047).E.coli was the most common pathogen cultured from the three sam-ples.Compared with midstream urine and pelvic urine,a greater proportion of E.coli in stone were resistant to aminoglycoside and β-lactamase antibiotics.Conclusion Positive bacterial culture for stone was associated with an increased incidence of post-operative SIRS.Bacterial culture for stone might have important clinical value in the treatment of postoperative infection in FURL patients.

Objective:To investigate the efficacy and safety of mini-track,mini-nephroscopy and mini-ultrasonic probe percutaneous nephrolithotomy(3mPCNL)for the treatment of 1.5-2.5 cm kidney stones.Methods:The perioperative data and postoperative follow-up data of a total of 25 patients with about 1.5-2.5 cm kidney stones who underwent 3mPCNL under ultrasound guidance in Peking Univer-sity People's Hospital from November 2023 to January 2024 were retrospectively analyzed.During the matching period,the 25 patients with 1.5-2.5 cm kidney stones receiving standard percutaneous neph-rolithotomy(sPCNL)were matched one-to-one according to the criterion that the absolute difference of the maximum diameter of stones between the two groups was less than 1 mm.The operative time,renal function changes,postoperative stone-free rate,hemoglobin changes,and complication rate of the two treatments were compared,and then the effectiveness and safety of 3mPCNL were preliminarily analyzed.Results:There were no significant differences in mean age,preoperative median creatinine,preoperative mean hemoglobin,preoperative mean hematocrit,median stone maximum diameter,and median stone CT density between the 3mPCNL group and the sPCNL group.The median operation time in the 3mPCNL group was 60.0(45.0-110.0)min,with no statistical significance compared with the sPCNL group,and all the patients underwent single-channel operations.The mean hemoglobin after operation in the 3mPCNL group was(115.3±15.5)mmol/L,and there was no significant difference between the preoperative group and the sPCNL group,and the mean hemoglobin decreased significantly between the sPCNL group and the sPCNL group[(9.5±2.2)mmol/L vs.(10.1±1.9)mmol/L].The mean hematocrit after operation was(28.0±5.2)％,and the difference was statistically significant compared with that before operation(t=2.414,P=0.020).The mean hematocrit drop was not statistically signi-ficant compared with the sPCNL group(2.3％vs.2.7％).The median serum creatinine in the 3mPCNL group was 74.0(51.0-118.0)μmol/L after operation,and the difference was statistically significant compared with that before operation(Z=-2.980,P=0.005).The stone-free rate in the 3mPCNL group and the sPCNL group was 96.0％and 97.3％,respectively,and the mean hospital stay was(4.3±1.4)d and(5.5±2.0)d,respectively,with the statistical significance(t=0.192,P=0.025).After the operation,one patient in sPCNL group had massive hemorrhage after the nephrostomy tube was re-moved,which was improved after selective renal artery embolization.One patient in the 3mPCNL group developed mild perirenal hematoma,which was improved after conservative treatment,and no complica-tions were observed in the other patients.Conclusion:3mPCNL in the treatment of 1.5-2.5 cm kidney stones can achieve an effective rate comparable to sPCNL,and can achieve the ideal stone-free rate in a shorter operative time with a lower rate of surgery-related complications.

Humans ; COVID-19 ; SARS-CoV-2

Objective:To evaluate the value of CT for differentiating gastric leiomyoma (GLM) from gastric schwannoma (GS).Methods:The clinical and imaging data of 42 patients with GLM (GLM group) and 41 patients with GS (GS group) were analyzed retrospectively. The general information and CT features were compared between two groups. The independent factors for differentiating GLM from GS were obtained by multivariate Logistic regression analysis. The receiver operating characteristic curve (ROC) was used to evaluate the diagnostic efficiency of the model.Results:The proportion of female and age in GLM group were significantly lower than those in GS group: 59.52% (25/42) vs. 85.37% (35/41), (51.83 ± 10.52) years old vs. (58.80 ± 10.63) years old, and there were statistical differences ( P<0.01). The upper part of the stomach rate, irregular shape rate, intraluminal growth rate, ratio of long diameter to short diameter and mild to moderate enhancement rate in GLM group were significantly higher than those in GS group: 71.43% (30/42) vs. 14.63% (6/41), 52.38% (22/42) vs. 21.95% (9/41), 92.86% (39/42) vs. 19.51% (8/41), 1.90 ± 0.55 vs. 1.34 ± 0.28 and 92.86% (39/42) vs. 51.22% (21/41), the cystic degeneration rate, ulcer rate, incidence of tumor-associated lymph node, CT values of venous phase and delayed phase in GLM group were significantly lower than those in GS group: 2.38% (1/42) vs. 26.83% (11/41), 7.14% (3/42) vs. 24.39% (10/41), 2.38% (1/42) vs. 60.98% (25/41), (59.21 ± 9.75) HU vs. (66.22 ± 10.33) HU and (65.02 ± 8.62) HU vs. (76.85 ± 11.89) HU, and there were statistical differences ( P<0.01 or <0.05); there were no statistical difference in the rate of calcification and the CT values of plain scan and arterial phase between the two groups ( P>0.05). Multivariate Logistic regression analysis result showed that the tumor location, growth mode, tumor-associated lymph node and ratio of long diameter to short diameter were the independent factors for differentiating GLM from GS ( OR = 34.385, 25.314, 0.023 and 97.700; 95% CI 2.848 to 415.171, 2.674 to 239.670, 0.001 to 0.637 and 3.113 to 3 066.549; P<0.01 or <0.05); when the model threshold was >0.647, the area under the curve was 0.988 (95% CI 0.934 to 1.000), with a sensitivity of 92.9% and specificity of 97.6%. Conclusions:When the tumor is prone to the upper part of the stomach, intraluminal growth, ratio of long diameter to short diameter >1.28, and the absence of the tumor-associated lymph node, GLM tends to be considered, on the contrary, it tends to be GS. Therefore, CT imaging features have certain value in differentiating GLM from GS before surgery.