ObjectiveTo investigate the target proteins directly bound by neochlorogenic acid （NA） and the molecular mechanisms that ameliorate the proliferation and inflammatory response of psoriatic keratinocytes. MethodsM5-induced HaCaT cells were used as a psoriatic keratinocyte proliferation and inflammatory cell model. The synthesized NA probe （NA-P） and NA prodrug were first evaluated for cell viability using a cell proliferation/cell counting kit-8（CCK-8）. The potency of NA and NA-P was evaluated in the safe concentration range， and the effects of 0-100 μmol·L^-1 NA and probe on M5-induced proliferation of HaCaT cells were detected using CCK-8. The effects of 20， 40， 80 μmol·L^-1 NA and 80 μmol·L^-1 NA-P on the expression of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， interleukin-23 （IL-23）， and interleukin-17A （IL-17A） inflammatory factors were detected by enzyme-linked immunosorbent assay （ELISA）， and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to measure the effects of NA on the mRNA expression of keratin 16 （K16） in HaCaT cells， S100 calcium-binding protein A9 （S100A9）， S100 calcium-binding protein A7 （S100A7）， IL-6， IL-17A， and chemokine 1 （CXCL1）. In vitro fluorescence labeling and competition experiments using NA-P were performed， and target protein angling and analysis using pull-down experiments combined with liquid chromatography-mass spectrometry （Pull-down/LC-MS/MS） were conducted. Target validation was performed using pull-down experiments combined with protein immunoblotting （Pull down-WB）， cellular heat transfer analysis combined with protein immunoblot （CETSA-WB） experiments， and molecular docking. Finally， Real-time PCR was utilized to detect the effects of 20， 40， 80 μmol·L^-1 NA and 80 μmol·L^-1 NA-P on the mRNA expression of IL-1β， nucleotide-binding oligomeric structural domain-like receptor protein 3 （NLRP3）， apoptosis-associated speckled-like protein （ASC）， and cysteine protease-1 （Caspase-1） in HaCaT cells. Protein immunoblot （Western blot） was used to detect the effects of phosphorylated p65 （p-p65）， p65， phosphorylated human nuclear factor-κB inhibitory protein α （p-IκBα）， human nuclear factor κB inhibitory protein α （IκBα）， and heat shock protein 90 （HSP90） expression. ResultsIn the 200 μmol·L^-1 safe concentration range， HaCaT cell proliferation， increased expression of TNF-α， IL-1β， IL-23， and IL-17A inflammatory factors， and increased mRNA expression of K16， S100A9， S100A7， IL-6， IL-17A， and CXCL1 were observed in the M5 group compared with the blank group. Cell proliferation in 5-100 μmol·L^-1 NA and NA-P groups was inhibited， and the expression of TNF-α， IL-1β， IL-23， and IL-17A inflammatory factors was decreased in the NA-L， NA-M， NA-H， and NA-P-H groups. The mRNA expression of K16， S100A9， S100A7， IL-6， IL-17A， and CXCL1 was decreased （P<0.05）. High-confidence targets were screened for HSP90 protein by Pull-down/LC-MS/MS using 200 μmol·L^-1 NA competing with 100 μmol·L^-1 NA-P. Compared with that in the blank group， the mRNA expression of NLRP3， IL-1β， ASC， and Caspase-1， as well as the expression of p-p65/p65， p-IκBα/IκBα， and HSP90 protein， were increased in HaCaT cells in the M5 group （P<0.05）. Compared with that in the M5 group， the mRNA expression of NLRP3， IL-1β， ASC， and Caspase-1 of cells in the NA-L group， the NA-M group， the NA-H group， and the NA-P-H group was decreased （P<0.05）. p-p65/p65 and p-IκBα/IκBα were decreased in the NA-M and NA-H groups （P<0.05）， and there was no change in HSP90 protein. Pull down-WB showed that NA could directly target HSP90 protein， and NA binding to HSP90 protein enhanced its thermal stability. Molecular docking of NA with HSP90 family proteins HSP90AA1， HSP90B1， and HSP90AB1 all resulted in highly stable binding. ConclusionNA can inhibit the proliferation and inflammatory response of psoriatic keratinocytes by a mechanism that may be achieved by targeting HSP90 to modulate the NF-κB/NLRP3 signaling pathway.

ObjectiveTo investigate the target proteins directly bound by neochlorogenic acid （NA） and the molecular mechanisms that ameliorate the proliferation and inflammatory response of psoriatic keratinocytes. MethodsM5-induced HaCaT cells were used as a psoriatic keratinocyte proliferation and inflammatory cell model. The synthesized NA probe （NA-P） and NA prodrug were first evaluated for cell viability using a cell proliferation/cell counting kit-8（CCK-8）. The potency of NA and NA-P was evaluated in the safe concentration range， and the effects of 0-100 μmol·L^-1 NA and probe on M5-induced proliferation of HaCaT cells were detected using CCK-8. The effects of 20， 40， 80 μmol·L^-1 NA and 80 μmol·L^-1 NA-P on the expression of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， interleukin-23 （IL-23）， and interleukin-17A （IL-17A） inflammatory factors were detected by enzyme-linked immunosorbent assay （ELISA）， and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to measure the effects of NA on the mRNA expression of keratin 16 （K16） in HaCaT cells， S100 calcium-binding protein A9 （S100A9）， S100 calcium-binding protein A7 （S100A7）， IL-6， IL-17A， and chemokine 1 （CXCL1）. In vitro fluorescence labeling and competition experiments using NA-P were performed， and target protein angling and analysis using pull-down experiments combined with liquid chromatography-mass spectrometry （Pull-down/LC-MS/MS） were conducted. Target validation was performed using pull-down experiments combined with protein immunoblotting （Pull down-WB）， cellular heat transfer analysis combined with protein immunoblot （CETSA-WB） experiments， and molecular docking. Finally， Real-time PCR was utilized to detect the effects of 20， 40， 80 μmol·L^-1 NA and 80 μmol·L^-1 NA-P on the mRNA expression of IL-1β， nucleotide-binding oligomeric structural domain-like receptor protein 3 （NLRP3）， apoptosis-associated speckled-like protein （ASC）， and cysteine protease-1 （Caspase-1） in HaCaT cells. Protein immunoblot （Western blot） was used to detect the effects of phosphorylated p65 （p-p65）， p65， phosphorylated human nuclear factor-κB inhibitory protein α （p-IκBα）， human nuclear factor κB inhibitory protein α （IκBα）， and heat shock protein 90 （HSP90） expression. ResultsIn the 200 μmol·L^-1 safe concentration range， HaCaT cell proliferation， increased expression of TNF-α， IL-1β， IL-23， and IL-17A inflammatory factors， and increased mRNA expression of K16， S100A9， S100A7， IL-6， IL-17A， and CXCL1 were observed in the M5 group compared with the blank group. Cell proliferation in 5-100 μmol·L^-1 NA and NA-P groups was inhibited， and the expression of TNF-α， IL-1β， IL-23， and IL-17A inflammatory factors was decreased in the NA-L， NA-M， NA-H， and NA-P-H groups. The mRNA expression of K16， S100A9， S100A7， IL-6， IL-17A， and CXCL1 was decreased （P<0.05）. High-confidence targets were screened for HSP90 protein by Pull-down/LC-MS/MS using 200 μmol·L^-1 NA competing with 100 μmol·L^-1 NA-P. Compared with that in the blank group， the mRNA expression of NLRP3， IL-1β， ASC， and Caspase-1， as well as the expression of p-p65/p65， p-IκBα/IκBα， and HSP90 protein， were increased in HaCaT cells in the M5 group （P<0.05）. Compared with that in the M5 group， the mRNA expression of NLRP3， IL-1β， ASC， and Caspase-1 of cells in the NA-L group， the NA-M group， the NA-H group， and the NA-P-H group was decreased （P<0.05）. p-p65/p65 and p-IκBα/IκBα were decreased in the NA-M and NA-H groups （P<0.05）， and there was no change in HSP90 protein. Pull down-WB showed that NA could directly target HSP90 protein， and NA binding to HSP90 protein enhanced its thermal stability. Molecular docking of NA with HSP90 family proteins HSP90AA1， HSP90B1， and HSP90AB1 all resulted in highly stable binding. ConclusionNA can inhibit the proliferation and inflammatory response of psoriatic keratinocytes by a mechanism that may be achieved by targeting HSP90 to modulate the NF-κB/NLRP3 signaling pathway.

BACKGROUND: Stroke is the leading cause of death and long-term disability worldwide, including China. This study aimed to provide timely updates on stroke burden and stroke-related risk factors to help improve population-based prevention and control strategies. METHODS: Based on the Global Burden of Disease study 2021, incidence rate, prevalence rate, mortality rate, and disability-adjusted life-year (DALY) rate were used to estimate stroke burden trend from 1990 to 2021. RESULTS: In 2021, China had 4.1 million incident stroke cases, 26.3 million prevalent stroke cases, 2.6 million stroke related deaths, and 53.2 million stroke related DALYs, compared to 11.9 million incident stroke cases, 93.8 million prevalent stroke cases, 7.3 million stroke related deaths, and 160.5 million stroke-related DALYs worldwide. In 2021, the top six risk factors contributing to stroke burden were high blood pressure, air pollution, tobacco consumption, dietary risk factors, high low-density lipoprotein cholesterol, and high fasting plasma glucose, both in China and worldwide. From 1990 to 2021, China had significant increases of incidence rate, prevalence rate, mortality rate, and DALY rate for stroke, with estimates of 100.6 (95% uncertainty intervals [UI]: 87.2, 114.1)%, 102.9 (95% UI: 95.5, 110.9)%, 40.0 (95% UI: 14.9, 72.3)% and 15.7 (95% UI: -4.6, 41.2)%, respectively, while global incidence rate, prevalence rate, mortality rate and DALY rate for total stroke showed relatively moderate increases or even decreases, with estimates of 15.0 (95% UI: 12.1,18.0)%, 25.8 (95% UI: 23.7, 28.0)%, -2.6 (95% UI: -10.6, 5.5)%, and -10.7 (95% UI: -17.7, -3.6)%, respectively. CONCLUSION Stroke remains a huge disease burden worldwide and in China, and compared to the worldwide China has a significantly higher burden of stroke.
Humans ; Stroke/etiology* ; China/epidemiology* ; Risk Factors ; Global Burden of Disease ; Disability-Adjusted Life Years ; Prevalence ; Incidence ; Female ; Quality-Adjusted Life Years ; Male

Objective To evaluate the clinical value of transarterial catheterization C-arm CT perfusion scanning technique during prostatic artery embolization(PAE)for benign prostatic hyperplasia(BPH).Methods The clinical data of 46 patients with BPH received PAE were analyzed retrospectively.All patients underwent prostatic artery(PA)digital subtraction angiography(DSA)and C-arm CT perfusion scanning to identify PA and prevent non-target organ embolization.The final recognization of PA was consulted by three senior doctors.After C-arm CT confirmation,PA was embolized with 100-300 μm polyvinyl alcohol(PVA)particles or microspheres under fluoroscopy.The postoperative complications and 3-month clinical efficacy were observed.Results A total of 106 vessels were angioraphed in 46 patients,with 83 PA vessels and 23 non-PA vessels.PA was identified by DSA and C-arm CT with sensitivity of 81.9%(68/83)and 100%(83/83),respectively,which showed significance(χ2=22.3,P<0.01).Non-PA was identified by DSA and C-arm CT with specificity of 73.9%(17/23)and 100%(23/23),which showed significance(χ2=9.2,P=0.02).No serious complications were observed and 3-month clincial efficacy was 91.3%.Conclusion Transarterial catheterization C-arm CT perfusion scanning technique can accurately identify PA,reduce PA leakage and prevent non-target organ embolization.

Objective To evaluate the efficacy of microwave ablation(MWA)synchronously with biopsy for pulmonary nodules.Methods The data of 64 patients with MWA combined with biopsy were analyzed retrospectively.Thirty-one patients(non-synchronous group)were treated with ablation following biopsy in turn to identify malignant tumors,and 33 patients(synchronous group)were treated by ablation and biopsy synchronously.The technical success rate,operation time,complications,hospitalization time and expenses were compared between non-synchronous group and synchronous group.Results The technical success rate,pneumothorax,and pleural effusion rate showed no significance between the two groups(P>0.05).There were all significant differences in operation time(42.00 min vs 54.26 min),hospitalization time(5.09 days vs 9.26 days),hospitalization expenses(26 840.61 yuan vs 32 527.26 yuan),lung hemorrhage(27.27%vs 87.10%)and hemoptysis(3.03%vs 19.35%)between synchronous group and non-synchronous group,respectively(P<0.05).Conclusion MWA synchronously with biopsy for pulmonary nodules is safe and feasible,which can reduce intraoperative bleeding,shorten treatment period and reduce hospitalization expenses.

To establish a simple,convenient,sensitive,and specific method for rapid detection of Zi-ka virus(ZIKV),the whole genome sequences of ZIKV isolated from different times and regions were analyzed.The specific primers and probes were designed based on the screened target se-quences located in the conserved region of the ZIKV NS5 gene.By combining RT-LAMP isother-mal amplification technology and immunochromatography technology,a reverse transcription loop mediated isothermal amplification nucleic acid and flow visualization strip(RT-LAMP-VF)detec-tion method for ZIKV was established.The results showed that the method had good specificity and sensitivity.When the ratio of inner,outer,and ring primers(FIP∶LF∶F3)was 4∶2∶1,the detection method can specifically detect 102 copies/pL RNA transcripts or 2.15 pfu ZIKV at 61 ℃for 45 minutes,with no cross reaction with other flaviviruses such as Japanese encephalitis virus and classical swine fever virus.Other RNAs in blood tissue samples did not affect the sensitivity and specificity of RT-LAMP-VF,indicating that the method can be applied to clinical practice.The ZIKV RT-LAMP-VF detection method established in this study is easy to perform and does not require special instruments and equipment.It is particularly suitable for the rapid detection of ZIKV in grassroots units,providing technical support and material support for the establishment of on-site rapid detection and early warning and prediction systems for ZIKV disease.

Traditional Chinese medicine is a treasure of Chinese civilization,which has undergone thousands of years of precipitati-on and clinical verification.According to the theory of traditional Chinese medicine,ancient doctors used natural medicines and natural means to promote self-regulation and damage repair of the body.In recent years,as an important tool to repair body damage,stem cells have set off a research upsurge.With the continuous deepening of stem cell research and the continuous expansion of the fields in-volved,the potential correlation between traditional Chinese medicine and stem cells has attracted more and more domestic and foreign researchers to invest in research.This paper expounds regenerative medicine and stem cells,the advantages of Chinese medicine in regulating stem cells,and the opportunities and challenges faced by Chinese medicine and regenerative medicine,and reviews the cur-rent research trends and frontier trends in this field,in order to provide useful references for subsequent studies.

Objective:To investigate different regulatory effects of S100A8/A9 expressed by keratinocytes in 3 common inflammatory skin injury modes: UVB-induced skin injury, allergic contact dermatitis, and psoriasis.Methods:Wild-type C57BL/6JGpt mice aged 6 to 8 weeks were selected for the following experiments: (1) mouse models of UVB-induced skin injury were established by single exposure to ultraviolet B (UVB) radiation on the shaved dorsal skin of mice (UVB group, n = 4), with the mice receiving no UVB radiation serving as a control group ( n = 4) ; (2) mouse models of allergic contact dermatitis were established by application of 2,4-dinitrochlorobenzene (DNCB) to the right ears of mice (DNCB group, n = 4), with the left ears of mice treated with a vehicle control serving as a control group ( n = 4) ; (3) mouse models of psoriasis-like skin inflammation were established by topical application of imiquimod cream to the depilated dorsal skin of mice (imiquimod group, n = 4), with the mice treated with vaseline serving as a control group ( n = 4). Hematoxylin-eosin (HE) staining was performed to assess histopathological changes in mouse skin tissues obtained from each group, and immunohistochemical study and Western blot analysis were performed to determine the expression of S100A8 and S100A9 in the mouse dorsal epidermis or ear skin lesions. In vitro cultured HaCaT cells were subjected to the following experiments: (1) cells in the UVB group were treated with a single UVB irradiation at a dose of 50 mJ/cm 2, and cells in the control group received no irradiation; (2) some cells were treated with tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) (collectively referred to as TI), and named as the TI group, which simulated the inflammatory environment in allergic contact dermatitis, while cells treated with corresponding solvents served as the control group; (3) cells were treated with 5 cytokines (interleukin 17A [IL-17A], IL-22, IL-1α, oncostatin M, and TNF-α, collectively referred to as M5), and named as the M5 group, which simulated the inflammatory environment in psoriasis, while cells treated with corresponding solvents served as the control group. Real-time fluorescence-based quantitative PCR, Western blot analysis, and enzyme-linked immunosorbent assay were performed to determine the mRNA and protein expression of S100A8 and S100A9, and to detect the extracellular secretion level of S100A8/A9, respectively. Results:Immunohistochemical study and Western blot analysis revealed that S100A8 and S100A9 expression levels were significantly higher in the skin lesions of mouse models of UVB-induced skin injury, allergic contact dermatitis, and psoriasis-like skin inflammation than in their corresponding control groups; immunohistochemical study further demonstrated that the increase in the expression of the two proteins was more pronounced in the mouse models of psoriasis-like skin inflammation. In the in vitro cell experiments, the mRNA expression of S100A8 and S100A9 in HaCaT cells at 12 and 24 hours were markedly higher in the UVB group (e.g., at 24 hours, 6.14 ± 0.60 vs. 1.00 ± 0.08, 2.58 ± 0.06 vs. 1.02 ± 0.22, respectively, both P < 0.01), TI group (e.g., at 24 hours, 3.90 ± 0.75 vs. 1.00 ± 0.02, 2.42 ± 0.30 vs. 1.01 ± 0.13, respectively, both P < 0.05), and M5 group (e.g., at 24 hours, 157.59 ± 9.30 vs. 1.00 ± 0.11, 251.37 ± 6.63 vs. 1.00 ± 0.03, both P < 0.001) than in the corresponding control groups, so were the extracellular secretion levels of S100A8/A9 at 24 and 48 hours (all P < 0.001), with some differences observed in their response patterns; notably, the response was more pronounced in the mouse model of psoriasis-like skin inflammation. Additionally, the protein expression levels of S100A8 and S100A9 in HaCaT cells were significantly higher in the M5 group than in the control group ( t = 4.66, 4.63, respectively, both P < 0.01), but were significantly lower in the UVB group ( t = -3.75, -3.34, P = 0.020, 0.029, respectively) and TI group ( t = -3.30, -4.50, P = 0.030, 0.011, respectively) than in the control groups. Conclusion:Keratinocytes exhibited active responses following 3 common inflammatory skin injuries, with their effector molecules S100A8 and S100A9, as damage-associated molecular patterns, playing crucial roles in UVB-induced skin injury, allergic contact dermatitis, and psoriasis, and the response seemed to be more pronounced in psoriasis-like dermatitis.

Objective To explore the efficiency and safety of trans-sheath intraluminal forceps biopsy under digital subtraction angiography(DSA)guidance for assisting diagnosis of pulmonary artery obstructive diseases.Methods Data of 16 patients who underwent trans-sheath intraluminal forceps biopsy for pulmonary artery obstructive diseases were retrospectively analyzed,and the clinical manifestations were recorded.The technical success of biopsy was defined as tissue obtained met the needs of pathology diagnosis.For patients with malignant pathology results,the final diagnosis was malignant,for those with benign pathology results after biopsy and no obvious changes after 6-month or longer follow-up,or benign pathology results after surgical resection,the final diagnosis was benign,otherwise was no clear diagnosis.The operation time,technical success rate,diagnostic efficiency,complications and changes of pulmonary artery pressure before and after the biopsy were observed.Results Among 16 patients,9 complained of intermittent chest tightness,4 complained of chest pain with chest tightness,2 complained of chest pain but 1 denied any symptoms.The lesions located in the left lung in 10 cases and in the right lung in 6 cases,all with enhanced CT showed filling defects of the involved branch of pulmonary artery.Totally 16 trans-sheath intraluminal forceps biopsies were performed in 16 patients,with an average operation time of(31.02±6.02)min and technical success rate of 100%.Malignant tumors were finally diagnosed in 10 cases,including 1 case of lung cancer with false-negative biopsy result,while biopsy correctly diagnosed benign lesions in the other 6 cases.Transient worsening chest pain with chest tightness occurred in 2 cases and relieved after symptomatic treatments.No statistically significant difference of pulmonary artery pressure was found before([53.38±14.28]mmHg)and after([53.69±14.15]mmHg)biopsy(P＞0.05).Conclusion DSA-guided trans-sheath intraluminal forceps biopsy was relatively safe and valuable for assisting diagnosis of pulmonary artery obstructive diseases.

Objective To observe the feasibility and effectiveness of fluoroscopy-guided Fustar adjustable bent sheath clamp biopsy for diagnosing obstructive esophageal diseases.Methods Totally 29 patients with esophageal or esophagogastric junction obstruction who failed to complete endoscopic biopsy were retrospectively analyzed.Real-time fluoroscopy-guided clamp biopsy of lesion areas were performed through 10F Fustar adjustable bent sheath under local anesthesia.The technical success rate,operation time,radiation dose were recorded,and the complications were evaluated.Results Clamp biopsy of lesion areas were successfully performed in all 29 cases,with technical success rate of 100%(29/29),the average operation time of(29.81±10.05)min and the average radiation dose of(127.14±100.36)mGy.No serious complication such as esophageal perforation nor massive bleeding occurred.After biopsy,22 cases(22/29,75.86%)were preliminarily diagnosed as positive,among them 2 cases underwent surgical operation,and the postoperative pathological results were consistent with biopsy.Negative biopsy results were found in 7 cases(7/29,24.14%),among them 2 cases underwent clamp biopsy again 3 months later which showed positive results.Conclusion Fustar adjustable bent sheath clamp biopsy was feasible and effective for diagnosing obstructive esophageal diseases,which could be regarded as the substitution and supplementation of endoscopic clamp biopsy.