ObjectiveTo investigate the material basis of the pathogenesis of cerebral ischemic injury with blood stasis and toxin syndrome and to explore the protective effects of Huayu Jiedu prescription （HYJDP） on neutrophil extracellular trap-related cell death （NETosis） in cerebral ischemic injury following acute cerebral infarction. MethodsSeventy-two Sprague-Dawley （SD） rats were randomly divided into six groups （n=12 per group）： sham operation （Sham） group， blood stasis and toxin model （Model） group， low-， medium-， and high-dose HYJDP groups （HYJDP-L， HYJDP-M， and HYJDP-H； 9， 18， and 36 g·kg^-1， respectively）， and butylphthalide （NBP） group （0.06 g·kg^-1）. Except for the Sham group， rats in all other groups were subjected to carrageenan/dry yeast combined with a modified intraluminal filament method to establish a focal cerebral ischemia model of the middle cerebral artery with blood stasis and toxin syndrome. Neurological function was evaluated at 24 h after modeling using the Zea-Longa neurological deficit score. Cerebral infarction rate was assessed by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Pathological morphology of brain tissue was observed using hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was used to determine serum levels of interleukin-8 （IL-8）， myeloperoxidase-DNA complexes （MPO-DNA）， and citrullinated histone H3 （CitH3）. Protein expression of phosphorylated phosphatidylinositol 3-kinase （p-PI3K）， protein kinase B （p-Akt）， mammalian target of rapamycin （p-mTOR）， sequestosome 1 （p62）， and CitH3 in brain tissue was detected by Western blot. Immunofluorescence （IF） was used to detect the expression of neutrophil-specific marker Ly6G， CitH3， and neuron-specific nuclear protein （NeuN） in brain tissue. ResultsCompared with the Sham group， neurological deficit scores and cerebral infarction rates in the model group were significantly increased （P<0.01 for both）. HE staining showed varying degrees of neuronal degeneration and necrosis， characterized by blurred neuronal structures， nuclear pyknosis and fragmentation， cytoplasmic dissolution into a vacuolated reticular pattern， and mild glial cell proliferation. ELISA results showed that serum levels of IL-8， MPO-DNA， and CitH3 were significantly increased （P<0.01）. Western blot analysis demonstrated decreased expression of p-PI3K， p-Akt， p-mTOR， and p62， while CitH3 expression was significantly increased （P<0.01）. IF results showed an increased number of NETs⁺ cells and a significant decrease in NeuN⁺ cells （P<0.01）. Compared with the Model group， neurological deficit scores in the HYJDP-H group were significantly decreased （P<0.05）， and cerebral infarction rates in the HYJDP-H and NBP groups were significantly reduced （P<0.01）. HE staining showed that brain tissue damage was markedly alleviated in the HYJDP-H group. ELISA results showed that levels of IL-8， MPO-DNA， and CitH3 were significantly decreased in the HYJDP-M， HYJDP-H， and NBP groups （P<0.01）. Western blot analysis showed that expression of p-PI3K， p-Akt， p-mTOR， and p62 was significantly increased in the HYJDP-H and NBP groups， while CitH3 expression was significantly reduced in all drug intervention groups （P<0.01）. IF results showed that the number of NETs⁺ cells was significantly decreased and the number of NeuN⁺ cells was significantly increased in all drug intervention groups （P<0.01）. ConclusionNETs may be the material basis of the pathogenesis of cerebral ischemic injury characterized by blood stasis and toxin. HYJDP can regulate the PI3K/Akt/mTOR signaling pathway， reduce the release of pro-inflammatory mediators and NETosis-related products， alleviate cerebral ischemic injury caused by autophagy-dependent NETosis， and thereby exert a neuroprotective effect.

Objective·To detect the expression level of Fas-associated protein with death domain(FADD)in head and neck squamous cell carcinoma(HNSCC)and to explore the molecular mechanisms by which FADD promotes the proliferation of HNSCC cells.Methods·The GEPIA 2 database was utilized to analyze the expression level of FADD in tumor tissues and to evaluate its association with prognosis.Immunohistochemistry staining(IHC)was performed on HNSCC tissues to investigate the changes in FADD expression levels in normal,dysplastic,and tumor tissues.Stable FADD-knockdown Fadu and HSC3 cell lines were constructed and validated using Western blotting and quantitative real-time PCR(qRT-PCR).The regulatory effect of FADD on the proliferation of HNSCC cells was explored using the LiveCyte live-cell tracking system,colony formation assay,and cell viability assay.Proteins interacting with FADD were identified by co-immunoprecipitation mass spectrometry(Co-IP/MS),and further mechanistic studies were conducted using CRISPR/Cas9 technology,LiveCyte live-cell tracking system,and Western blotting.Results·Analysis of the GEPIA2 database indicated that FADD was significantly overexpressed in head and neck cancer and was associated with poor prognosis.IHC staining showed that FADD expression levels progressively increased from normal to dysplastic to tumor tissues in HNSCC patients.Knockdown of FADD in HNSCC cells resulted in significantly reduced proliferation and colony formation compared to the control group.Co-IP/MS results showed that FADD interacted with the CUX1 protein,and FADD knockdown led to increased CUX1 expression.Moreover,CUX1 knockdown significantly promoted HNSCC cell proliferation and reversed the anti-proliferative phenotype caused by FADD knockdown.Conclusion·FADD plays a significant pro-carcinogenic role in HNSCC and is associated with poor prognosis.FADD can further regulate tumor cell proliferation by interacting with CUX1 and suppressing its expression level.

Objective·To detect the expression level of Fas-associated protein with death domain(FADD)in head and neck squamous cell carcinoma(HNSCC)and to explore the molecular mechanisms by which FADD promotes the proliferation of HNSCC cells.Methods·The GEPIA 2 database was utilized to analyze the expression level of FADD in tumor tissues and to evaluate its association with prognosis.Immunohistochemistry staining(IHC)was performed on HNSCC tissues to investigate the changes in FADD expression levels in normal,dysplastic,and tumor tissues.Stable FADD-knockdown Fadu and HSC3 cell lines were constructed and validated using Western blotting and quantitative real-time PCR(qRT-PCR).The regulatory effect of FADD on the proliferation of HNSCC cells was explored using the LiveCyte live-cell tracking system,colony formation assay,and cell viability assay.Proteins interacting with FADD were identified by co-immunoprecipitation mass spectrometry(Co-IP/MS),and further mechanistic studies were conducted using CRISPR/Cas9 technology,LiveCyte live-cell tracking system,and Western blotting.Results·Analysis of the GEPIA2 database indicated that FADD was significantly overexpressed in head and neck cancer and was associated with poor prognosis.IHC staining showed that FADD expression levels progressively increased from normal to dysplastic to tumor tissues in HNSCC patients.Knockdown of FADD in HNSCC cells resulted in significantly reduced proliferation and colony formation compared to the control group.Co-IP/MS results showed that FADD interacted with the CUX1 protein,and FADD knockdown led to increased CUX1 expression.Moreover,CUX1 knockdown significantly promoted HNSCC cell proliferation and reversed the anti-proliferative phenotype caused by FADD knockdown.Conclusion·FADD plays a significant pro-carcinogenic role in HNSCC and is associated with poor prognosis.FADD can further regulate tumor cell proliferation by interacting with CUX1 and suppressing its expression level.

Objective To analyze the research trends and frontier of TCM regulation of NF-κB;To provide reference for related research.Methods Relevant literature about TCM regulation of NF-κB was retrieved from CNKI,VIP,Wanfang Data,CBM and Web of Science from January 1,2013 to December 31,2022.VOSviewer 1.6.19 and CiteSpace 6.2.R4 software were used for visualization analysis on authors,institutions and keywords.Results Totally 3 728 articles in Chinese and 995 in English were included,and the number of articles was on the rise.The Chinese and English articles involved 487 and 237 core authors,respectively,forming research teams represented by Yan Guanghai,Liu Jian,Wang Li,and Li Wei,Zhang Li,Zhang Yu,etc.There were 7 and 8 effective clusters in Chinese and English articles respectively.Keyword analysis showed that this research field mainly focused on diseases(inflammatory diseases,tumors,cardiovascular and cerebrovascular diseases,etc.),research methods(in vivo experiment,in vitro experiment)and intervention methods(acupuncture,TCM monomer,TCM compound,etc.).Conclusion TCM regulation of NF-κB mainly focuses on related diseases and intervention methods,and it is a research trend to find drug action targets and conduct experimental verification through network pharmacology and molecular docking technology.

This article introduces the concept, classification and application of microneedle in the field of cosmetology. It summarizes and analyzes the use of microneedle in promoting skin graft and flap survival, scar repair, facial rejuvenation, local anesthesia, hair loss treatment, skin relaxation, acne treatment, reducing color sink and other aspects, and looks forward to the bright prospects of microneedle applications in plastic surgery.

An analytical method was developed for the determination of five carbohydrate impurities in amino acid drug substances by high performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD). Sugar impurities in the amino acid sample were separated and enriched by cation exchange resin. A Lichropher NH2 column (4.6 mm × 250 mm, 5 μm) was used for chromatographic separation, and a gradient elution was performed using acetonitrile-water as mobile phase. The drift tube temperature was 40 oC, the gain value was 8, and nitrogen (350 kPa) was auxiliary gas. Method validation results showed that the limits of detection for fructose, glucose, sucrose, maltose and lactose were in the range of 20.8-75.0 mg/kg and that the limits of quantitation were in the range of 96.2-238.8 mg/kg. Good linear relationship (r ≥ 0.999) were in the linear range for the five sugars, and the recoveries ranged from 84.9%-107.8%. With easy operation, high sensitivity, good precision and reliable accuracy, the method can be used for analysis of residual sugar impurities in amino acid drug bulk drug.

Objective:To evaluate the safety and efficacy of combination of transjugular intrahepatic portosystemic shunt (TIPS) combined with interventional therapy for primary hepatic carcinoma complicated with portal hypertension.Methods:From October 2013 to December 2017, medical records of 141 patients with primary hepatic carcinoma complicated with portal hypertension hospitalized and treated in Anhui Provincial Cancer Hospital were collected. According to the inclusion and exclusion criteria, 32 cases were in the TIPS combined with interventional treatment group and 29 cases were included in the intervention-only group after age, gender, mean tumor diameter and Child-Pugh classification matched with TIPS combined with intervention treatment group. The efficacy of TIPS was obsented, and the related complications and prognosis of the two groups at six, 12 and 24 months after treatment were compared. Independent sample t test, Mann-Whitney U test and Chi-square test were used for statistical analysis, and Kaplan-Merier method was used for survival analysis. Results:There were no statistically significant differences between TIPS combined with intervention group and intervention-only group in age, gender, mean tumor diameter, Child-Pugh classification, Child-Turcotte-Pugh (CTP) score, Barcelona staging, interventional treatment, causes of liver cirrhosis, portal hypertension related symptoms , portal vein tumor thrombus, alanine aminotransferase (ALT), and total bilirubin (TBil) (all P>0.05). The success rate of TIPS of patients in TIPS combined with intervention group was 100% (32/32). All the portal hypertension related symptoms after TIPS were relieved, and the remission rate was 100% (32/32). Compared with that before TIPS, after TIPS, the portal vein pressure decreased ((36.5±6.9) cmH 2O vs. (25.5±5.6) cmH 2O (1 cmH 2O=0.098 kPa)), the diameter of portal vein decreased ((15.9±3.5) mm vs. (13.7±2.7) mm), and ascites volume decreased (abdominal circumference ((105.6±13.9) cm vs. (88.0±9.8) cm), red blood cell count ((2.6±0.8)×10 12/L vs. (3.3±1.3)×10 12 /L) and hemoglobin level ((78.9±15.5) g/L vs. (108.4±14.6) g/L) both increased, and the differences were statistically significant ( t=2.866, 3.105, 10.769, -3.548 and -3.869, all P<0.01). The stent patency rate six months after TIPS was 100% (32/32), the 12-month patency rate was 94% (30/32), and the 24-month patency rate was 84% (27/32). Six months, 12 months and 24 months after treatment, the incidence of ascites of patients in the TIPS combined with interventional treatment group was 0, 3.1% (1/32) and 9.4% (3/32), respectively, and the incidence of bleeding was 3.1% (1/32), 9.4% (3/32) and 15. 6% (5/32), respectively, which were all lower than those of the intervention-only group (13.8%, 4/29; 27.6%, 8/29; 65.5%, 19/29; 20.7%, 6/29; 34.5%, 10/29 and 62.1%, 18/29), and the differences were statistically significant ( χ2=4.72, 7.24, 20.80; and 4.62, 5.72, 13.97; all P<0.05). There were no statistically significant differences in the incidence rates of hepatic encephalopathy 6 and 12 months after treatment between the two groups (both P>0.05). The incidence of hepatic encephalopathy of intervention-only group (48.3%, 14/29) 24 months after treatment was higher than that of TIPS combined with intervention group (21.9%, 7/32), and the difference was statistically significant ( χ2=4.70, P=0.030). The results of Kaplan-Merier analysis indicated that 12 and 24 months after treatment the cumulative survival rates of TIPS combined with intervention group (84.4%, 27/32 and 53.1%, 17/32) were both higher than those of the intervention-only group (41.4%, 12/29 and 13.8%, 4/29), and the differences were statistically significant ( χ2=12.20 and 5.06, both P<0.05). The median survival time of TIPS combined with intervention group was 25 months, which was longer than that of the intervention-only group (12 months), and the difference was statistically significant ( Z=3.341, P=0.001). Conclusions:TIPS combined with interventional therapy is safe and effective in the treatment of primary hepatic carcinoma complicated with portal hypertension, which can improve the quality of life and increase the survival rate.

Objective:To investigate the influences of anxiety on postoperative efficacy in elderly patients with complex proximal humerus fractures.Methods:A retrospective case control study was conducted to analyze the clinical data of 297 elderly patients with complex proximal humerus fractures admitted to 5 tertiary hospitals in Shanghai from January 2017 to December 2018. There were 121 males and 176 females, aged 65-85 years [(74.2±8.5)years]. The fracture types included Neer type III in 176 patients and Neer type IV in 121 patients. Two days after admission, patients were analyzed by self-rating anxiety scale (SAS) and anxiety factors questionnaire (participated by Shanghai Mental Health Center) and they were divided into anxiety group (SAS>35) and nonanxiety group (SAS≤35). Anxiety group included 68 patients, among which 7 had conservative treatments. Nonanxiety group contained 229 patients, among which 7 had conservative treatments. Both groups underwent with proximal reduction of humerus fractures accepted the operations. Operative patient in both groups underwent open reduction and plate fixation. The surgery rate and length of hospital stay were compared between groups. After 1 year of follow-up, Constant-Murley score was used to evaluate the effect of shoulder joint and dual-energy X-ray absorptiometry (DXA) was taken to observe the wrist joint bone mineral density. Postoperative complications were recorded.Results:All operative patients were followed up for 10-28 months, with an average of 13.6 months. The surgery rate was 89.7% (61/68) in anxiety group and 96.9% (222/229) in nonanxiety group ( P<0.05). The hospital stay was (6.8±3.4)days for non-surgery patients and (10.2±2.8)days for surgery patients in anxiety group and (4.2±1.6)days for non-surgery patients and (7.4±2.2)days for surgery patients in nonanxiety group ( P<0.05). In anxiety group and nonanxiety group, the excellent and good rate of Constant-Murley score was respective 75.4% (46/61) and 88.3% (196/222) and the wrist joint bone mineral density was respective (-3.1±0.5)SD and (-2.9±0.6)SD (both P<0.05). The incidence of complications was 8.2% (5/61) in anxiety group and 4.5% (10/222) in nonanxiety group ( P>0.05). Conclusions:Anxiety has a significant negative influence on the surgery rate, length of hospital stay, shoulder joint function, wrist joint bone mineral density. Intervention on anxiety in elderly patients with complex proximal humerus fractures should be valued attention by clinical orthopedic surgeons.

Objective To evaluate the efficacy of transjugular intrahepatic portosystemic shunt (TIPS)in the treatment of patients with hepatic sinusoidal obstruction syndrome (HSOS).Methods From April 2015 to August 2018,at The First Affiliated Hospital of University of Science and Technology of China,21 patients with gynura segetum caused HSOS were selected.All the patients received TIPS treatment because of unresponsiveness to anticoagulant therapy for at least two weeks.After operation patients were followed up with liver and portal vein Doppler ultrasonography examination,liver and kidney function tests,and survival observation.T test,logistic univariate regression analysis and Cox regression analysis were performed for statistical analysis.Results Among the 21 patients with gynura segetum-related HSOS,18 patients were in the subacute phase and three patients in the chronic phase.All of them were moderate or severe patients and all successfully underwent TIPS.The postoperative portal vein pressure was (16.71 ± 4.68) cmH2O (1 cmH2O =0.098 kPa),which was lower than that before operation ((41.52 ±6.27) cmH2O),and the difference was statistically significant (t =16.936,P ＜ 0.01).The postoperation portal vein blood flow velocity was (41.52 ±7.70) cm/s,which was higher than before operation ((11.19 ± 3.29) cm/s),and the difference was statistically significant (t =-15.191,P ＜0.01).At one month after operation,15 of 21 patients were clinically cured;among the remaining six patients,four patients were improved and two patients were ineffective (including one patient died).At four months after operation,two patients died,and the remaining 19 patients were clinically cured.At one month after operation,the levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),total bilirubin (TBil) and serum creatinine were (23.7 ± 16.8) U/L,(33.9 ±7.4) U/L,(52.7 ± 38.2) μmol/L and (62.7 ± 12.6) μmol/L,respectively,which were lower than those before operation ((60.5 ± 42.4) U/L,(78.4 ± 42.4) U/L,(74.9 ± 38.2) μmol/L and (82.4 ± 19.6) μmol/L,respectively),and the differences were statistically significant (t =3.193,3.493,2.378 and 4.519;all P ＜ 0.05).The level of albumin was (39.0 ±3.1) g/L,which was higher than that before operation ((30.9 ± 3.8) g/L),and the difference was statistically significant (t =-10.283,P ＜ 0.01).Portal vein thrombosis and preoperative TBil level had predictive value for therapeutic efficacy (both P ＜0.05).The one-year cumulative survival rate of patients was 90.5％.Preoperative TBil level and hepatic encephalopathy had effects on the prognosis of patients (both P ＜ 0.05).Conclusion TIPS is a safe,reliable and effective treatment for patients with subacute and chronic gynura segetum-related HSOS who are not responding to ineffective anticoagulant therapy,which can improve the prognosis and survival rate of the patients.

Objective: To investigate the expression of immunomarkers CK7, CK20, CK17, CDX2, MUC1 and MUC2 in primary adenocarcinoma of the ampulla of Vater, to explore the role of these markers in the histopathologic subclassification of ampullary carcinoma; and to provide biologic basis for precision treatment of patients with different types of ampullary carcinoma. Methods: Forty-two cases of primary ampullary carcinoma were collected at Peking University People′s Hospital, from 2012 to 2018 year. There were 22 males and 20 females. Aged range 42 to 88 years old, with mean aged (62±11) years. Among the patients, 6 was high differentiation, 19 median differentiation, and 17 low differentiation. Immunohistochemical studies on the expression of CK7, CK20, CK17, CDX2, MUC1 and MUC2 were performed in 42 cases of primary ampullary carcinoma. The relationship between different ampullary carcinoma subtypes and clinicopathologic survival data was analyzed using SPSS 16.0 statistical software. Results: Three histopathologic subtypes were observed. Among 42 cases, 8(19.0%)were classified as intestinal subtype, which showed a positive expression rate of 8/8 for both CK20 and CDX2, and 5/8 for MUC2. Both CK7 and CK17 were weakly expressed in one case (1/8). No expression was observed for MUC1 in this subtype. Twenty-two (52.4%,22/42) cases were classified as pancreaticobiliary subtype, which showed a positive expression rate of 100.0%(22/22) for both CK7 and MUC1, and 90.9% (20/22) for CK17. No expression was observed for CK20, CDX2 and MUC2.The remaining 12 (28.6%) cases were classified as mixed subtype, which showed variable expression patterns. The expression frequencies of these 6 immunomarkers in different subtypes of ampullary carcinoma did not correlate with various clinicopathologic factors such as patient gender and age, tumor size, histologic differentiation, pancreatic and bile duct invasion, or the depth of duodenal invasion. However, stage Ⅲ+Ⅳ diseases were more commonly seen in pancreaticobiliary type (63.6%,14/22) than intestinal type (2/8) and mixed type (3/9; χ²=6.508, P=0.039). Follow-up data showed a trend of better survival rate for patients with intestinal subtype than those with mixed and pancreaticobiliary subtypes. Conclusions Ampullary carcinoma can be subclassified into three different subtypes using a panel of six immunomarkers, especially for the identification of subtypes of poorly differentiated carcinoma. CK7, CK17 and MUC1 are major markers of pancreaticobiliary subtype, whereas CK20, CDX2 and MUC2 are useful markers for intestinal subtype. The mixed subtype variably expresses these markers. The prognosis of patients with intestinal subtype appears better than that of pancreaticobiliary and mixed subtypes. Accurate subtyping of ampullary carcinoma is clinically important to patient management and prognosis assessment.