ObjectiveTo explore the effects and mechanism of the modified Danggui Yinzi on "itch-anxiety" model rats of chronic urticaria （CU）. MethodsThe 36 SPF-grade 6-8-week-old female SD rats were randomly divided into a blank control group，a model group，a positive control group，a low-dose modified Danggui Yinzi group，a medium-dose modified Danggui Yinzi group，and a high-dose modified Danggui Yinzi group. A "itch-anxiety" model was established by intraperitoneal injection of a suspension of sodium chloride and aluminum hydroxide and ovalbumin，combined with chronic unpredictable emotional stress stimulation. After successful modeling，rats in each group were administered drugs by gavage. The positive control group was given intragastric administration of the drug solutions of cetirizine and fluoxetine （2.08 mg·kg^-1·d^-1 fluoxetine， 2 mg·kg^-1·d^-1 cetirizine）， the low-，medium-，and high-dose modified Danggui Yinzi groups were administered traditional Chinese medicine at 1.44，2.88， 5.76 g·kg^-1， respectively，while the blank control group and model group were given an equal volume of normal saline. All interventions lasted for 15 days. Behavioral changes were evaluated by the elevated plus-maze test （detecting the percentage of entries into the open arms （OE%），the percentage of time spent in the open arms （OT%），and the total number of entries into the open and closed arms （TNE）），the open-field test （detecting total activity，average movement speed，and latency to enter the central area），and scratching behavior observation. Pathological changes of skin tissues were observed by hematoxylin-eosin （HE） staining and toluidine blue staining，while those of amygdala tissues were observed by HE staining，Nissl staining，and immunofluorescence detection of ionized calcium-binding adapter molecule-1 （Iba-1）. The content of immunoglobulin E （IgE），interleukin-33 （IL-33），histamine in serum and glutamate in the amygdala was detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the protein expression of striatal-enriched protein tyrosine phosphatase （STEP），N-methyl-D-aspartate receptor subunit 2B （NR2B）， calmodulin-dependent protein kinase Ⅱ （CaMKⅡ），phosphorylated CaMKⅡ （p-CaMKⅡ），mitogen-activated protein kinase （MAPK），phosphorylated MAPK （p-MAPK），nuclear factor kappa-light-chain-enhancer of activated B cells （NF-κB），phosphorylated NF-κB （p-NF-κB），and postsynaptic density protein-95 （PSD-95） in the amygdala. ResultsCompared with the blank control group，the model group rats showed obvious anxiety-like behaviors （decreased OE%，OT%，and TNE，reduced total activity，slower average movement speed，and prolonged latency to enter the central area），increased scratching times，obvious skin inflammation and mast cell degranulation，severe amygdala tissue damage，increased glutamate content in the amygdala，and elevated levels of IgE and IL-33 in serum. The expression of STEP，NF-κB，p-NF-κB，NR2B，MAPK，p-MAPK，CaMKⅡ，and p-CaMKⅡ proteins in the amygdala increased，while the expression of PSD-95 protein decreased （P<0.05）. Compared with the model group，the modified Danggui Yinzi group of each dose had increased OE%，OT%，TNE，total activity，and average movement speed，shortened latency to enter the central area， reduced scratching times，alleviated skin inflammation and mast cell degranulation，relieved amygdala tissue damage，decreased glutamate content in the amygdala，and reduced levels of IgE and IL-33 in serum. Moreover，compared with the model group，the low -，medium-，and high-dose modified Danggui Yinzi groups showed decreased expression levels of STEP，NF-κB，p-NF-κB，NR2B，MAPK，p-MAPK，CaMKⅡ，and p-CaMKⅡ proteins in the amygdala，and increased expression of PSD-95 protein. There was a significant dose-effect relationship，with the high-dose group showing the most significant regulatory effect （P<0.05）. ConclusionThe modified Danggui Yinzi has a therapeutic effect on "itch-anxiety" model rats of CU. Its mechanism may be related to regulating glutamate metabolism in the amygdala，modulating the STEP/NR2B/CaMKⅡ/MAPK/NF-κB pathway，and regulating the expression of PSD-95.

Objective To investigate the effect of knockdown PRTN3 on polarization phenotype and anti-tumor function of macrophages in breast cancer.Methods A mouse monocyte/macrophage cell line(RAW264.7)with knockdown of PRTN3 was constructed.The mRNA and protein levels of PRTN3 were detected by RT-qPCR and flow cytometry.Macrophages were co-cultured with tumor cells,and the polarization phenotypes of macrophages were detected by flow cytometry,and the mRNA levels of polarization and cytokines related genes were detected by RT-qPCR.The tumor phagocytosis and tumor killing ability of macrophages were determined by in vitro phagocytosis assay and tumor killing assay.The effect of macrophages with PRTN3 knockdown on tumor progression and metasta-sis was compared in an orthotopic transplantation model of mouse breast cancer.Results The mRNA and protein levels of PRTN3 in the RAW264.7 cell line with stable knockdown of PRTN3 were significantly decreased(P＜0.01).Knockdown of PRTN3 in RAW264.7 cells upregulated the expression of M1 phenotype-related genes(CD80,CD86,iNOS,IL-1β,IL-6 and TNF-α)(P＜0.05).After macrophages co-culture with tumor cells,macrophages with knockdown of PRTN3 upregulated the expression of M1 phenotype-related genes(CD80,CD86,iNOS,IL-1β,IL-6 and TNF-α)(P＜0.01).Knockdown of PRTN3 enhanced the phagocyto-sis(P＜0.05)and tumor killing ability(P＜0.05)of macrophages in vitro.Adoptive transfusion of PRTN3-knockdown macrophages into tumor-bearing mice inhibited the growth of in situ breast cancer and reduced lung metastasis(P＜0.05).Conclusions PRTN3 regulates phenotypic polarization of macrophages,and knockdown of PRTN3 enhances the anti-tumor function of macrophages.

Objective:Through microscopic and ultrastructural observations,to explore the origin of the cartilage of the anteroposterior pseudocyst.Staging for the auricular pseudocyst according to the different pathological changes and exploring its clinical significance.Methods:48 cases(51 ears)of the auricular pseudocyst were collected as the observation objects(46 males, 2 females, 15-76 years old) and 45 patients(45 ears)chronic suppurative otitis media hospitalized patients who underwent tympanic membrane repair as a normal control group(43 males, 2 females, 12-74 years old).The outer wall of the cyst was surgically resected and the residual auricular cartilage was retained in the control group for HE staining or transmission electron microscopy specimens, and the specimens were observed and analyzed under light microscope and transmission electron microscopy.Results:According to the data of this study, auricular pseudocyst was relatively common in people aged 21-60 years. The disease course of 4 ears was ≤ 10 days,the course of 16 ears was > 10 days yet ≤ 30 days, the course of 20 ears was > 1 month but ≤ 2 months,the course of 6 ears was > 2 months yet ≤ 1 year, the course of 3 ears was > one year but ≤ two years,and the course of 2 ears was > 2 years. The observation of microstructure and ultrastructure of the different course of outer walls of the auricle pseudocyst was shown:due to various causes of cartilage fluid that separated perichondrium and cartilage tissue, when the perichondrium was stimulated, the bone progenitor cells from the quiescence phase came to the activities period. With the extension of the course, the periosteum was thickening gradually, cartilage the bone progenitor cells idifferentiated into chondroblasts, which idifferentiated into chondrocytes last, from native to mature, from thin to thick. It was stable a period of time when the cartilage mature. The outer wall of the cyst was new cartilage, which reflected additional growth. The nucleus of the chondrocytes evolved into nucleolysis, and necrosis, when the lesions was stimulated.The control group had a thin layer of cartilage and periosteum, containing a small amount of osteoprogenitor cells; the cartilage layer was thick, the cartilage cells were small near the periosteum, and the deep cartilage cells were large, all of which were mature chondrocytes.Conclusions:The outer cartilage of auricle pseudocyst is newborn cartilage. The pathological staging is divided into early stage(cartilage formation), middle stage(cartilage maturity) and late stage(cartilage necrosis).This stage can provide a reference for exploring the pathogenesis of pseudocysts of the auricle and formulating surgical treatment principles.

Objective:Through microscopic and ultrastructural observations,to explore the origin of the cartilage of the anteroposterior pseudocyst.Staging for the auricular pseudocyst according to the different pathological changes and exploring its clinical significance.Methods:48 cases(51 ears)of the auricular pseudocyst were collected as the observation objects(46 males, 2 females, 15-76 years old) and 45 patients(45 ears)chronic suppurative otitis media hospitalized patients who underwent tympanic membrane repair as a normal control group(43 males, 2 females, 12-74 years old).The outer wall of the cyst was surgically resected and the residual auricular cartilage was retained in the control group for HE staining or transmission electron microscopy specimens, and the specimens were observed and analyzed under light microscope and transmission electron microscopy.Results:According to the data of this study, auricular pseudocyst was relatively common in people aged 21-60 years. The disease course of 4 ears was ≤ 10 days,the course of 16 ears was > 10 days yet ≤ 30 days, the course of 20 ears was > 1 month but ≤ 2 months,the course of 6 ears was > 2 months yet ≤ 1 year, the course of 3 ears was > one year but ≤ two years,and the course of 2 ears was > 2 years. The observation of microstructure and ultrastructure of the different course of outer walls of the auricle pseudocyst was shown:due to various causes of cartilage fluid that separated perichondrium and cartilage tissue, when the perichondrium was stimulated, the bone progenitor cells from the quiescence phase came to the activities period. With the extension of the course, the periosteum was thickening gradually, cartilage the bone progenitor cells idifferentiated into chondroblasts, which idifferentiated into chondrocytes last, from native to mature, from thin to thick. It was stable a period of time when the cartilage mature. The outer wall of the cyst was new cartilage, which reflected additional growth. The nucleus of the chondrocytes evolved into nucleolysis, and necrosis, when the lesions was stimulated.The control group had a thin layer of cartilage and periosteum, containing a small amount of osteoprogenitor cells; the cartilage layer was thick, the cartilage cells were small near the periosteum, and the deep cartilage cells were large, all of which were mature chondrocytes.Conclusions:The outer cartilage of auricle pseudocyst is newborn cartilage. The pathological staging is divided into early stage(cartilage formation), middle stage(cartilage maturity) and late stage(cartilage necrosis).This stage can provide a reference for exploring the pathogenesis of pseudocysts of the auricle and formulating surgical treatment principles.

Objective:To investigate the effects of the histone deacetylase(HDAC)inhibitor panobinostat on melanoma growth,tumor immunity,and the underlying mechanisms.Methods:B16F0 melanoma cells were cultured and treated with different concentrations of panobinostat.The effect of panobinostat on HDAC expression in B16F0 cells was detected by WB.The effects of panobinostat on the proliferation,migration,invasion,apoptosis and cell cycle of B16F0 cells were detected by CCK-8 assay,wound-healing assay,Transwell assay and flow cytometry,respectively.The effect of panobinostat on gene expression in B16F0 cells was detected using transcriptome analysis and verified by qPCR.Flow cytometry was used to detect the effect of panobinostat on the expression of MHCⅠ/Ⅱ in B16F0 cells.B16F0 cells and bone marrow-derived dendritic cells(BMDC)were co-cultured to assess the effects of panobinostat on the expression of CD11c,CD80 and CD86 in BMDC cells.A xenograft mouse model was used to evaluate the effects of panobinostat on tumor growth and host immune function.Results:panobinostat promoted the acetylation of H3 and α-tubulin proteins in B16F0 cells(P<0.01 or P<0.001 or P<0.000 1),inhibited cell proliferation,migration,and invasion,promoted apoptosis,and induced G1-phase cell cycle arrest(P<0.05 or P<0.001 or P<0.000 1).Additionally,panobinostat enhanced surface expression of MHC Ⅰ/Ⅱ on B16F0 cells and promoted BMDC maturation(all P<0.01).Transcriptomic analysis showed that panobinostat upregulated the expression of E-cadherin and antigen presentation related genes in B16F0 cells,and inhibited the expression of N-cadherin,vimentin,c-Myc and CDK1,which was confirmed by qPCR.In vivo,panobinostat suppressed xenograft tumor growth and enhanced immune function in tumor-bearing nude mice(P<0.05,P<0.000 1).Conclusion:panobinostat can inhibit the malignant biological behavior of B16F0 cells,promote apoptosis,regulate tumor immunity,and enhance immune function of tumor-bearing nude mice.

BACKGROUND: Cancer stem-like cells (CSCs) are a small subset of cells in tumors that exhibit self-renewal and differentiation properties. CSCs play a vital role in tumor formation, progression, relapse, and therapeutic resistance. B7-H3, an immunoregulatory protein, has many protumor functions. However, little is known about the mechanism underlying the role of B7-H3 in regulating gastric cancer (GC) stemness. Our study aimed to explore the impacts of B7-H3 on GC stemness and its underlying mechanism. METHODS: GC stemness influenced by B7-H3 was detected both in vitro and in vivo . The expression of stemness-related markers was examined by reverse transcription quantitative polymerase chain reaction, Western blotting, and flow cytometry. Sphere formation assay was used to detect the sphere-forming ability. The underlying regulatory mechanism of B7-H3 on the stemness of GC was investigated by mass spectrometry and subsequent validation experiments. The signaling pathway (Protein kinase B [Akt]/Nuclear factor erythroid 2-related factor 2 [Nrf2] pathway) of B7-H3 on the regulation of glutathione (GSH) metabolism was examined by Western blotting assay. Multi-color immunohistochemistry (mIHC) was used to detect the expression of B7-H3, cluster of differentiation 44 (CD44), and Nrf2 on human GC tissues. Student's t -test was used to compare the difference between two groups. Pearson correlation analysis was used to analyze the relationship between two molecules. The Kaplan-Meier method was used for survival analysis. RESULTS: B7-H3 knockdown suppressed the stemness of GC cells both in vitro and in vivo . Mass spectrometric analysis showed the downregulation of GSH metabolism in short hairpin B7-H3 GC cells, which was further confirmed by the experimental results. Meanwhile, stemness characteristics in B7-H3 overexpressing cells were suppressed after the inhibition of GSH metabolism. Furthermore, Western blotting suggested that B7-H3-induced activation of GSH metabolism occurred through the AKT/Nrf2 pathway, and inhibition of AKT signaling pathway could suppress not only GSH metabolism but also GC stemness. mIHC showed that B7-H3 was highly expressed in GC tissues and was positively correlated with the expression of CD44 and Nrf2. Importantly, GC patients with high expression of B7-H3, CD44, and Nrf2 had worse prognosis ( P = 0.02). CONCLUSIONS B7-H3 has a regulatory effect on GC stemness and the regulatory effect is achieved through the AKT/Nrf2/GSH pathway. Inhibiting B7-H3 expression may be a new therapeutic strategy against GC.
Humans ; Cell Line, Tumor ; Neoplasm Recurrence, Local ; NF-E2-Related Factor 2/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Signal Transduction ; Stomach Neoplasms

BACKGROUND: Synovectomy has been introduced into total knee arthroplasty (TKA) with the aim of relieving pain and inflammation of the synovium. However, there are no long-term, comparative data to evaluate the effect of synovectomy in TKA. This study was aimed at assessing pain, function, and complications in patients undergoing synovectomy during TKA for osteoarthritis (OA) at long-term follow-up. METHODS: This was a prospective randomized controlled trial of 42 consecutive patients who underwent staged bilateral TKA. Patients undergoing the first-side TKA were allocated to receive TKA with or without synovectomy followed by a 3-month washout period and crossover to the other strategy for the opposite-side TKA. The overall efficacy of both strategies was evaluated by determination of blood loss, the Knee Society score (KSS), and knee inflammation conditions during a 3-month postoperative period. The postoperative pain, range of motion (ROM), and complications were sequentially evaluated to compare the two groups until 10 years after surgery. RESULTS: At the 10-year follow-up, both groups had a similarly significantly improved ROM (114.88 ± 9.84° vs. 114.02 ± 9.43°, t  = 0.221, P  = 0.815) and pain relief with no differences between the two groups (1.0 [1.0] vs. 1.0 [1.5], U  = 789.500, P  = 0.613). Similar changes in total blood loss, KSS, and knee inflammation were found in both groups during 3 months postoperatively ( P  > 0.05). Additionally, there was no significant difference regarding complications and satisfaction between the two groups ( P  > 0.05). CONCLUSIONS: Synovectomy in conjunction with TKA for primary OA does not seem to provide any benefit regarding postoperative pain, ROM, and satisfaction during a 10-year follow-up. In addition, it may not result in more blood loss and increased incidence of long-term complications. Based on our long-term findings, it should not be performed routinely. TRIAL REGISTRATION Chinese Clinical Trial Registry, ChiCTR-INR-16008245; https://www.chictr.org.cn/showproj.aspx?proj=13334 .
Humans ; Arthroplasty, Replacement, Knee/methods* ; Synovectomy/methods* ; Osteoarthritis, Knee/surgery* ; Prospective Studies ; Pain, Postoperative ; Inflammation/etiology* ; Range of Motion, Articular ; Knee Joint/surgery* ; Treatment Outcome ; Knee Prosthesis/adverse effects*

Objective:To explore the feasibility of high-flow nasal cannula(HFNC)therapy for respiratory failure in elderly patients.Methods:A total of 300 patients with respiratory failure admitted to Peking University Shougang Hospital from December 2016 to March 2019 were enrolled in this prospective study.Patients were divided into three groups: the HFNC group, the conventional oxygen therapy(COT)group and the non-invasive positive pressure ventilation(NPPV)group(n=100 in each group). Arterial oxygen saturation(SPO 2), oxygen index(OI), heart rate(HR), respiratory rate(RR), mean arterial pressure(MAP), comfort level, discharge rate, tracheal intubation rate, rate of referral to ICU, mortality and rate of referral to another group after therapy were compared between the HFNC and COT groups and between the HFNC and NPPV groups. Results:SPO 2 after oxygen therapy for 30 minutes( t=-2.992, P=0.003), 1 hour( t=-2.884, P=0.005)and 6 hours( t=-3.196, P=0.002)and OI before discharge( t=-2.060, P=0.048)were higher in the HFNC group than in the COT group.The HR in the above two groups was lower before discharge than before therapy, and the HR in the COT group was even lower(73.1±25.1 beat per minute vs.75.1±25.9 beat per minute), but both were within the normal range.The discharge rate was higher( χ2=-1.969, P=0.049), while the rate of referral to another group was lower in the HFNC group than in the COT group( χ2=-3.115, P=0.002). There was no significant difference in the tracheal intubation rate, ICU transfer rate and mortality between the HFNC and COT groups.SPO 2 after oxygen therapy for 30 minutes( t=-2.026, P=0.046)and 6 hours( t=-2.101, P=0.040)were higher in the HFNC group than in the NPPV group, but there was no significant difference in OI and SPO 2 between the two groups before discharge.The HR in both HFNC and NPPV groups was lower before discharge than before therapy, and there was no statistical difference between the two groups.The mortality, discharge rate, tracheal intubation rate, ICU transfer rate and rate of referral to another group had no significant difference between the HFNC and NPPV groups.The comfort level was higher in the HFNC group than in the COT and NPPV groups( t=-3.758 and -19.180, both P=0.000). Conclusions:HFNC is a new type of oxygen therapy equipment introduced after COT and NPPV, and possesses more advantages for elderly patients with respiratory failure.

As an income redistribution mechanism, the design of Urban Resident Basic Medical Insurance ( URBMI) should reflect the inclination to take care of vulnerable populations, such as people with chronic diseases and low incomes. Therefore, whether the healthcare services of vulnerable populations have been improved, is the most important indicator to determine the effectiveness of the URBMI. Using the DID model, this paper analyzes healthcare service utilization of invulnerable and vulnerable populations before and after the establishment of URBMI ( including both outpatient and inpatient) , based on the idea that these two populations have different socio-economic and health status. Then, based on the gap between the healthcare service utilization changes of different groups, we can measure the equity in URBMI. The results show that the establishment and implementation of URBMI has indeed narrowed the gap of invulnerable and vulnerable populations with different socio-economic and health status; health-care service utilization of vulnerable populations has increased significantly, and health status has significantly im-proved. From vertical and horizontal perspectives, the results prove that the establishment of URBMI has improved the equity of healthcare service utilization among different populations across China.

OBJECTIVETo formulate a novel histological typing and grading-rated system for colorectal cancer (CRC) for evaluating the biological behavior of CRC and prognosis.

METHODSAccording to the highly heterogeneous histological features, WHO classification and histological differentiation criteria, and other biological behavior parameters of CRC, a novel histological typing and grading-scale system for CRC was designed. The histological typing and corresponding grading-scale of CRC was defined as the following: (1) No mucin-producing adenocarcinoma, including tubular adenocarcinoma, sieve-like acne adenocarcinoma, medullary carcinoma, serrated adenocarcinoma and micropapillary carcinoma, etc. (1-3 points); (2) Mucin-producing adenocarcinoma, including mucinous adenocarcinoma and signet ring cell carcinoma (3-4 points); (3) Squamous cell carcinoma (1-3 points); (4) Neuroendocrine tumors, including neuroendocrine tumors, neuroendocrine carcinoma (1-4 points); (5) The special type of CRC, including clear cell carcinoma, spindle cell carcinoma, etc. (4-points); (6) Undifferentiated carcinoma (5 points). The pathology report form was formatted based on the major histological type with the secondary histological type. The final total score of CRC was defined as the sum of the corresponding grading scores for different histological types. The total score of a single-structure CRC was defined as the corresponding grading score multiplied by 2. A total of 666 patients with advanced CRC were pathologically reviewed and analyzed to assess the correlation of the histological typing and grading scores with TNM staging and lymph node metastasis.

RESULTSThe results showed a significant correlation of the histological grading-scale and TNM staging and lymph node metastasis (P<0.05). The scores of CRC histological grading-scale increased synchronously with the TNM staging and lymph node metastasis rate.

CONCLUSIONThe novel histological grading system allows objective evaluation of the biological behaviors and prognosis of CRC for determining individualized postoperative treatment. This system still needs further revision and updates based on evidence from prospective, multi-centered, large-scale trials.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Grading ; methods ; Neoplasm Staging ; Prognosis ; Prospective Studies ; Young Adult