OBJECTIVE To compare the consistency between the semi-quantitative detection of HPV E6/E7 mRNA and the detection of p16 IHC and E6/E7RNA ISH in the tissues,and the feasibility of detecting high-risk HPV in head and neck mucosal lesions by HPV E6/E7 mRNA detection in the swabs was discussed.METHODS A total of 100 cases of head and neck mucosal lesions treated by the Department of Head and Neck Surgery,Beijing Tongren Hospital Affiliated to Capital Medical University from September 2022 to August 2023 were collected.Semi-quantitative detection of HPV E6/E7 mRNA was performed in oropharynx,lesion surface swab and lesion tissue specimen,and p16 immunohistochemical staining(IHC)and E6/E7 mRNA in situ hybridization(ISH)were detected in lesion tissue,and the consistency and difference of different detection results were studied.RESULTS Among the 100 patients,83 met the inclusion criteria and were divided into 21 papilloma cases,10 polyps/chronic inflammation cases,19 laryngeal cancer cases,13 oropharyngeal cancer cases,and 20 hypopharyngeal cancer cases according to pathological diagnosis.The HPV E6/E7 mRNA semi-quantitative results of oropharyngeal swab and lesion surface swab showed moderate or near high consistency with p16 IHC results.The results of HPV E6/E7 mRNA semi-quantitative in diseased tissue were highly consistent with those of p16 IHC(Kappa=0.780).In the diagnostic efficacy analysis,both swabs showed high consistency with HPV E6/E7 mRNA ISH(Kappa=0.690 and 0.708).CONCLUSION In the head and neck mucosal lesions,the HPV semi-quantitative detection results of oropharyngeal and lesion surface swab showed good consistency compared with classical p16 IHC and gold standard HPV E6/E7 mRNA ISH.It is a simple and reliable method for clinical high-risk HPV detection,which is helpful for the screening and individualized precise prevention and control of HPV infection in head and neck mucosal lesions.

Immune checkpoint inhibitors (ICIs) have demonstrated unparalleled clinical responses and revolutionized the paradigm of tumor treatment, while substantial patients remain unresponsive or develop resistance to ICIs as a single agent, which is traceable to cellular metabolic dysfunction. Although dysregulated metabolism has long been adjudged as a hallmark of tumor, it is now increasingly accepted that metabolic reprogramming is not exclusive to tumor cells but is also characteristic of immunocytes. Correspondingly, people used to pay more attention to the effect of tumor cell metabolism on immunocytes, but in practice immunocytes interact intimately with their own metabolic function in a way that has never been realized before during their activation and differentiation, which opens up a whole new frontier called immunometabolism. The metabolic intervention for tumor-infiltrating immunocytes could offer fresh opportunities to break the resistance and ameliorate existing ICI immunotherapy, whose crux might be to ascertain synergistic combinations of metabolic intervention with ICIs to reap synergic benefits and facilitate an adjusted anti-tumor immune response. Herein, we elaborate potential mechanisms underlying immunotherapy resistance from a novel dimension of metabolic reprogramming in diverse tumor-infiltrating immunocytes, and related metabolic intervention in the hope of offering a reference for targeting metabolic vulnerabilities to circumvent immunotherapeutic resistance.
Humans ; Neoplasms/pathology* ; Immunotherapy/methods* ; Immune Checkpoint Inhibitors/therapeutic use*

ObjectiveTo investigate the Alzheimer-associated neurofilament protein （AD7c-NTP） in urine of middle-aged and elderly people and its correlation between common metabolites. MethodsA total of 1 150 middle-aged and elderly people who did their physical exmanination in the health examination center of the Sichuan Science City Hospital and the Third Hopital of Mianyang were recruited from March 2017 to March 2020. The level of urine AD7c-NTP were measured by enzyme-linked immunosorbent assay （ELISA）， and common metabolites in blood were measured by biochemical analyzer. Based on urine AD7c-NTP level ≤1.5 ng/mL， the objects was divided into normal group （n=956） and elevated group （n=194）. Thier demographic data and blood biochemical indicators were collected. ResultsThe urine AD7c-NTP level in middle-aged and elderly people was 0.60（0.30~1.20） ng/mL. The urine AD7c-NTP level was higher in women than that in men ［1.04（0.40~1.30） ng/mL vs. 0.84（0.30~1.00） ng/mL， Z=4.202， P˂0.01］. And the urine AD7c-NTP level was lower in the normal group than that in the elevated group ［0.50（0.30~0.90） ng/mL vs. 2.10（1.70~2.10） ng/mL， Z=22.035， P˂0.01］. The results of the univariate comparison showed that， the differences between the two groups in age （Z=6.545）， fasting glucose （Z=3.506）， blood uric acid （Z=2.574）， urea nitrogen （Z=2.891）， creatinine （Z=2.243）， total bilirubin （Z=3.936）， glutathione （Z=0.969）， total cholesterol （t=3.956） and low density lipoprotein （Z=-5.678） were were statistically significant （P˂0.05 or 0.01）. Spearman correlation analysis showed that， the urine AD7c-NTP level was positively correlated with age and the levels of urea nitrogen， glucose， total cholesterol and low density lipoprotein （r=0.177， 0.178， 0.171， 0.109， 0.149， P˂0.01）， and negatively correlated with the level of total bilirubin （r=-0.172， P˂0.01）. Conclusionthe urine AD7c-NTP level in middle-aged and elderly females was signifitcantly higher than in middle-aged and elderly males.The urine AD7c-NTP level of middle-aged and elderly people was positively correlated with age， urea nitrogen， glucose， total cholesterol and low density lipoprotein， and negatively correlated with total bilirubin.

Objective:To investigate the predictive value of platelet-to-neutrophil ratio (PNR) on hemorrhagic transformation (HT) and poor outcomes at 90 d after intravenous thrombolysis (IVT) in patients with acute ischemic stroke (AIS).Methods:Patients with AIS received IVT in Hefei Second People's Hospital from July 2019 to June 2021 were retrospectively enrolled. HT was defined as intracerebral hemorrhage found by CT reexamination within 24 h after IVT, and the poor outcome was defined as the modified Rankin Scale score ≥3 at 90 d after onset. Multivariate logistic regression analysis was used to determine the independent predictors of HT and poor outcomes at 90 d, and the predictive value of PNR on HT and poor outcomes at 90 d was analyzed by receiver operating characteristic (ROC) curve. Results:A total of 202 patients with AIS treated with IVT were included, of which 32 had HT and 50 had poor outcomes at 90 d after onset. Multivariate logistic regression analysis showed that PNR at 24 h after IVT was significantly and independently negatively correlated with the poor outcomes (odds ratio [ OR] 0.959, 95% confidence interval [ CI] 0.928-0.991; P=0.012); PNR at admission ( OR 0.886, 95% CI 0.827-0.948; P<0.001) and PNR at 24 h after IVT ( OR 0.923, 95% CI 0.879-0.969; P=0.001) were significantly independently and negatively correlated with HT. ROC curve analysis showed that the area under the curve of PNR at 24 h after IVT for predicting poor outcomes was 0.733 (95% CI 0.659-0.807; P=0.012), the best cutoff value was 35.03, and the predictive sensitivity and specificity were 70.4% and 74%, respectively. The area under the curve of PNR at admission for predicting HT was 0.830 (95% CI 0.774-0.886; P<0.001), the best cutoff value was 34.81, and the predictive sensitivity and specificity were 70% and 93.7%, respectively. The area under the curve of PNR at 24 h after IVT for predicting HT was 0.770 (95% CI 0.702-0.839; P=0.001), the best cutoff value was 41.73, and the predictive sensitivity and specificity were 53.5% and 96.9%, respectively. Conclusion:For patients with AIS treated with IVT, lower PNR at 24 h after IVT is an independent predictor of the poor outcomes at 90 d, while PNR at admission and 24 h after IVT are the independent predictors of HT.

Objective:To observe any effect of electroacupuncture (EA) on the expression of phosphorylated extracellular signal-regulated protein kinase (p-ERK1/2) and phosphorylated cyclic adenosine monophosphate response element binding protein (p-CREB) in the spinal dorsal horns of diabetics experiencing neuropathic pain.Methods:Eight rats were randomly selected from 30 healthy male Sprague-Dawley rats as the normal group (N), and the remaining twenty-two rats were treated with a single high-dose intraperitoneal injection of streptozotocin (STZ) to establish a neuropathic pain model. The rats modeled successfully were randomly divided into a model group (M, n=8) and an EA group ( n=8). In the EA group, electroacupuncture was applied at the bilateral Hou san li and Kunlun acupoints starting on the 15th day after the STZ injection. The daily sessions lasted 30 minutes for 1 week. Body weight (BW), fasting blood glucose (FBG) and paw withdrawal latency (PWL) were observed before the STZ injection and on the 7th, 14th, and 21st days afterward. The expression of p-ERK1/2 and p-CREB in the dorsal horns of the rats′ spinal cords was detected using western blotting. The count of p-CREB-positive cells in the dorsal horns and their co-localization with neurons was detected using immunofluorescence. Results:In comparison with the N group, the average BW of the M group on the 7th, 14th and 21st days after the STZ injection was significantly lower, while the average FBG was significantly higher. There was no significant difference between the M and N groups in the average PWL on the 7th day after the STZ injection, but it had decreased significantly in the M group on the 14th and 21st days. Compared with the M group, the average PWL of the EA group was significantly longer on the 21st day after the injection. The expression of p-ERK1/2 and p-CREB protein in the spines of the M group was significantly higher than in the N group. p-CREB positive cells were more numerous in the M group compared with the N group, while in the EA group they were fewer. P-CREB was co-located with neurons in the spinal dorsal horn.Conclusion:EA can alleviate neuropathic pain effectively, perhaps by inhibiting the expression of p-ERK1/2 and p-CREB in the dorsal horns of the spinal cord.

Depression is a common emotional disorder, which has high incidence rate and mortality worldwide. Ascorbic acid (AA) is also known as vitamin C. As a kind of reducing vitamin, AA participates in anti-oxidation, enzyme assistance and neuro-modulation in the central nervous system. Clinical evidence and animal studies show that AA has anti-depressant effect. This article reviews the function of AA in the nervous system, the current researches on relation between AA and depression, and possible mechanism, and analyzes its therapeutic significance in this disease, hoping to provide new ideas for the clinical treatment of depression.

Objective:To investigate the marital adjustment status of breast cancer patients after surgery and analyze its influencing factors.Methods:Using the convenient sampling method, postoperative breast cancer patients who were treated in 3 ClassⅢ Grade A hospitals in Shanghai from June 2017 to December 2018 were selected as the research objects. The general information questionnaire, Revised Dyadic Adjustment Scale (RDAS) , Breast Cancer Self-Efficacy Scale (BCSES) , Body Image Scale (BIS) , Hospital Anxiety and Depression Scale (HADS) and Medical Outcomes Study Social Support Survey (MOS-SSS) were used for investigation. Univariate analysis, Pearson correlation analysis and multiple linear regression were used to analyze the influencing factors of marital adjustment. In this study, a total of 300 questionnaires were issued and 240 valid questionnaires were returned. The effective recovery rate was 80.0%.Results:The total score of RDAS for postoperative breast cancer patients was (48.88±10.38) . The results of Pearson correlation analysis showed that marital adjustment of postoperative breast cancer patients was positively correlated with self-efficacy, body image, and medical and social support ( r=0.357, 0.382, 0.507; P<0.01) , and it was negatively correlated with anxiety and depression ( r=-0.362, -0.373; P<0.01) . The results of multiple linear regression analysis showed that awareness of tumor staging, actual support, self-acceptance and body image were the influencing factors of marital adjustment in patients after breast cancer surgery ( P<0.05) , explaining 32.9% of the total variation. Conclusions:The marital adjustment ability of patients after breast cancer surgery is at a moderate level. The awareness of tumor staging, actual support, self-acceptance and body image are factors that affect the patients' marital adjustment. It is necessary to actively formulate targeted intervention measures to enhance the psychological adjustment ability of patients in order to improve the ability of marriage adjustment.

Objective:To investigate the correlation between neutrophil to lymphocyte ratio (NLR) and 30-day clinical outcomes in patients with spontaneous cerebral hemorrhage and whether adding NLR to ICH score improve the accuracy of predicting poor outcomes.Methods:Patients with spontaneous intracerebral hemorrhage admitted to the Department of Neurology, the Second People's Hospital of Hefei from March 2018 to April 2019 were enrolled retrospectively. The demographic and baseline clinical and imaging data were documented. The absolute neutrophil counts and absolute lymphocyte counts within 24 h of onset were obtained and NLR was calculated. At 30 d after the onset of cerebral hemorrhage, the modified Rankin Scale was used to evaluate the outcomes. Good outcome was defined as ≤2, and poor outcome was defined as >2. Multivariate logistic regression analysis was used to determine the independent risk factors for poor outcomes. The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of baseline NLR, ICH score and NLR+ ICH score for poor outcomes in patients with spontaneous cerebral hemorrhage. Results:A total of 159 patients with spontaneous cerebral hemorrhage were enrolled, including 106 males (66.67%), age 62.29±15.10 years. Neutrophil count was (7.30±3.95)×10 9/L, lymphocyte count was (1.41±0.67)×10 9/L, and NLR was 6.94±7.66. Baseline hematoma volume was 17.93±25.87 ml, median ICH score 0 (interquartile range 0-1). The outcomes of 60 patients (37.7%) were poor at 30 d. Univariate analysis showed that coronary heart disease, systolic blood pressure, diastolic blood pressure, high-sensitivity C-reactive protein, fasting blood glucose, white blood cell count, neutrophil count, NLR, hematoma broken into the ventricle, hematoma volume, NIHSS and ICH scores in the poor outcome group were significantly higher than those of the good outcome group (all P<0.05). Multivariate logistic regression analysis showed that NLR was an independent predictor of poor outcomes at 30 d after the onset of spontaneous cerebral hemorrhage (odds ratio 1.135, 95% confidence interval 1.092-2.321; P=0.038). The ROC curve analysis showed that the best cut-off value of NLR was 6.679, and the sensitivity and specificity of predicting poor outcomes were 51.67% and 76.77% respectively; the best cut-off value of ICH score was 1.0, and the sensitivity and specificity of predicting poor outcomes were 69.71% and 89.80% respectively; the sensitivity and specificity of the combined application of NLR + ICH score to predict poor outcomes were 74.58% and 82.65% respectively. Conclusions:NLR was independently associated with poor outcomes at 30 d after the onset of spontaneous cerebral hemorrhage. Adding it to the ICH score could improve the accuracy of predicting poor outcome.

Objective To investigate the protective effect of Honokiol on the airway inflammation induced by particulate matter 2.5(PM2.5)in the asthmatic mice and its mechanism.Methods Fifty male specific pathogen free (SPF)Balb/c mice were randomly divided into 5 groups.Group A:normal control group;group B:asthmatic model group;group C:PM2.5 exposure asthmatic group;group D:TAK -242 group;group E:Honokiol group. Asthmatic mouse models were established by ovalbumin(OVA)sensitization and challenge.On days 0 and 7,the mice in B-E groups were injected intraperitoneally with injection 100 mg/L OVA and aluminum hydroxide for sensitization;on days 14 to 21,10 g/L OVA solution was given 30 min per day to challenge.During challenge phrase,the mice in C -E groups received intratracheal injection of PM2.5,every other day,4 times totally.On this basis,the mice in group D re-ceived TAK-242 intraperitoneal injection,and the mice in group E received honokiol intragastric administration.Group A was given saline instead of OVA.Animals were sacrificed 24 h after the final inhalation challenge,and the bronchoal-veolar lavage fluid(BALF)of the left lung was used for differential inflammatory cell counts.The expressions of Toll-like receptors 4(TLR4)and nuclear factor(NF)-κB at mRNA level were detected by real-time quantitative PCR. Flow cytometry analysis was performed to measure the levels of Th17 and Treg cells.Results Compared with group A,mice in group B and group C expressed more serious disorders of bronchial epithelial cells,alveolar wall congestion and edema,increased mucus secretion in the airway and infiltration of inflammatory cells in the lung,and those in group C were more obvious than those of group B and group E significantly reduced respiratory inflammation;compared with group A[(4.15 ± 1.35)×108/L,0.012 0 ± 0.002 3],the total number of inflammatory cell counts[(16.79 ± 5.62)×108/L and(24.58 ± 13.46)×108/L],eosinophils proportions(0.113 8 ± 0.022 3 and 0.197 8 ± 0.084 9)in group B and group C,were significantly higher,and the differences were statistically significant(all P<0.05);The total number of inflammatory cell counts and eosinophils proportion in group E(8.56 ± 3.28)×108/L and 0.041 5 ± 0.013 5)were significantly lower than those in group C,and the differences were statistically significant(all P <0.05);The expressions of TLR4 mRNA and NF-κB mRNA in group B and C(1.85 ± 0.56,1.82 ± 0.28 and 2.97 ± 0.41,2.83 ± 0.32)were significantly higher,and the differences were statistically significant(all P <0.05);The expressions of TLR4 mRNA and NF-κB mRNA in group E(1.60 ± 0.28,1.54 ± 0.25)was significantly lower than those in group C,and the differences were statistically significant(all P<0.05);the expressions of Th17 in group B and C[(2.89 ± 0.61)% and(4.96 ± 0.27)%]were significantly higher than those of group A[(1.03 ± 0.35)%] (all P<0.05);The expression of Th17 in group E[(1.83 ± 0.23)%]was significantly lower than that of group C,and the differences were statistically significant(P<0.05);the expressions of Treg in group B and C[(4.96 ± 0.35)%and(2.27 ± 0.41)%]were significantly lower than those of group A[(7.37 ± 0.56)%],and the differences were sta-tistically significant(all P<0.05);The expression of Treg in group E was significantly increased[(6.45 ± 0.38)%] compared with that in group C,and the difference were statistically significant(P<0.05);and those of group D and E were improved remarkably.Conclusions Honokiol can relieve PM2.5 exposure of asthmatic airway inflammation through down-regulating the expression of TLR4 and NF-κB and Th17 and regulating the balance of Th17 and Treg cells.

Objective To explore the effect of particulate matter (PM 2.5) on airway inflammation in asthmatic mice and the intervention effect of Honokiol.Methods Fifty female BALB/c mice were divided into 5 groups according random number table,group A:normal control group;group B:asthma model group;group C:PM 2.5 low dose exposure asthma group;group D:PM 2.5 high dose exposure asthma group:group E:Honokiol group.Asthmatic mouse models were established by ovalbumin(OVA) sensitization and challenge.On day 0 and 7,B-E groups were intraperitoneally with injection 100 mg/L OVA and Al (OH)3 for sensitization;on day 14 to 21,10 g/L OVA solution was given 30 min per day to challenge.During challenge phrase,C-D groups were received different doze intratracheal injection of PM 2.5 respectively,every 7 days,total 4 times.On this basis,the mice in group E received Honokiol intragastfic administration.The mice in group A were carried out by using saline instead of OVA.Mice were sacrificed 24 h after the final inhalation challenge,and for the recovered bronchoalveolarlavage fluid(BALF) of the left lung was used for differential inflammatory cell counts,HE staining and pathological examination were performed on the right lung.The expression of Toll-like receptor 4 (TLR4) and nuclear factor (NF)-κB at mRNA level were detected by real-time flurescence quantitative polymerase chain reaction (qPCR).Flow cytometry analysis was performed to measure the levels of Th17 and Treg cells.Results Compared with group A,mice in group B,group C and group D expressed more serious disorsers of bronchial epithelial cells,alveolar wall congestion and edema,increased mucus secretion in the airway and infiltration of inflammatory cells in lung,and those in group D was more obvious than those in group C and group E,significantly reduced respiratory inflammation compared with group E[(8.56 ± 3.28) × 108/L,0.041 5 ± 0.013 5],the total number of inflammatory cell counts in group C and group D were (20.28 ± 11.16) × 108/L and (27.38 ± 14.64) × 108/L,eosinophils proportion were 0.177 8 ±0.064 9 and 0.229 1 ±0.098 7,there were statistically significant differences(all P ＜ 0.05);compared with group E (1.60 ± 0.28,1.54 ± 0.25),the expression of TLR4 mRNA and NF-κB mRNA in group C and group D (2.56 ± 0.49,3.21 ± 0.61;2.42 ± 0.30,2.83 ± 0.32) were significantly higher,and there were statistically significant differences(all P ＜0.05),group D was more higher than those in group C (all P ＜ 0.05);compared with group E(0.018 3 ± 0.002 3),the expression of Th17 in group C and group D (0.043 9 ±0.008 9 and 0.052 2 ±0.011 8) were significantly higher,and there were statistically significant differences(all P ＜0.05);compared with group E(0.064 5 ±0.003 8),the expression of Treg in group C and group D (0.038 2 ± 0.004 2) and (0.022 7 ± 0.003 3) were significantly lower,and there were statistically significant differences(all P ＜ 0.05);and those of group E were improved remarkably.Conclusion PM 2.5 exposure can aggravate airway inflammation in asthmatic mice,and the damage to airway is more obvious when exposed to high dose of PM 2.5,Honokiol can relieve PM 2.5 exposure of asthmatic airway inflammation through down regulation the expression of TLR4 and NF-κB and Th17 and regulating the balance of Th17 and Treg cells.