ObjectiveTo investigate the influence of histone deacetylase 3 (HDAC3) on the occurrence, development of psoriasis-like inflammation in mice, and the relative immune mechanisms. MethodsHealthy C57BL/6 mice aged 6-8 weeks were selected and randomly divided into 3 groups: control group (Control), psoriasis model group (IMQ), and HDAC3 inhibitor RGFP966-treated psoriasis model group (IMQ+RGFP966). One day prior to the experiment, the back hair of the mice was shaved. After a one-day stabilization period, the mice in Control group was treated with an equal amount of vaseline, while the mice in IMQ group was treated with imiquimod (62.5 mg/d) applied topically on the back to establish a psoriasis-like inflammation model. The mice in IMQ+RGFP966 group received intervention with a high dose of the HDAC3-selective inhibitor RGFP966 (30 mg/kg) based on the psoriasis-like model. All groups were treated continuously for 5 d, during which psoriasis-like inflammation symptoms (scaling, erythema, skin thickness), body weight, and mental status were observed and recorded, with photographs taken for documentation. After euthanasia, hematoxylin-eosin (HE) staining was used to assess the effect of RGFP966 on the skin tissue structure of the mice, and skin thickness was measured. The mRNA and protein expression levels of HDAC3 in skin tissues were detected using reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB), respectively. Flow cytometry was employed to analyze neutrophils in peripheral blood and lymph nodes, CD4⁺ T lymphocytes, CD8⁺ T lymphocytes in peripheral blood, and IL-17A secretion by peripheral blood CD4⁺ T lymphocytes. Additionally, spleen CD4⁺ T lymphocyte expression of HDAC3, CCR6, CCR8, and IL-17A secretion levels were analyzed. Immunohistochemistry was used to detect the localization and expression levels of HDAC3, IL-17A, and IL-10 in skin tissues. ResultsCompared with the Control group, the IMQ group exhibited significant psoriasis-like inflammation, characterized by erythema, scaling, and skin wrinkling. Compared with the IMQ group, RGFP966 exacerbated psoriasis-like inflammatory symptoms, leading to increased hyperkeratosis. The psoriasis area and severity index (PASI) skin symptom scores were higher in the IMQ group than those in the Control group, and the scores were further elevated in the IMQ+RGFP966 group compared to the IMQ group. Skin thickness measurements showed a trend of IMQ+RGFP966>IMQ>Control. The numbers of neutrophils in the blood and lymph nodes increased sequentially in the Control, IMQ, and IMQ+RGFP966 groups, with a similar trend observed for CD4⁺ and CD8⁺ T lymphocytes in the blood. In skin tissues, compared with the Control group, the mRNA and protein levels of HDAC3 decreased in the IMQ group, but RGFP966 did not further reduce these expressions. HDAC3 was primarily located in the nucleus. Compared with the Control group, the nuclear HDAC3 content decreased in the skin tissues of the IMQ group, and RGFP966 further reduced nuclear HDAC3. Compared with the Control and IMQ groups, RGFP966 treatment decreased HDAC3 expression in splenic CD4⁺ and CD8⁺ T cells. RGFP966 treatment increased the expression of CCR6 and CCR8 in splenic CD4⁺ T cells and enhanced IL-17A secretion by peripheral blood and splenic CD4⁺ T lymphocytes. Additionally, compared with the IMQ group, RGFP966 reduced IL-10 protein levels and upregulated IL-17A expression in skin tissues. ConclusionRGFP966 exacerbates psoriatic-like inflammatory responses by inhibiting HDAC3, increasing the secretion of the cytokine IL-17A, and upregulating the expression of chemokines CCR8 and CCR6.

Objective·To investigate the efficacy and safety of hypertonic dextrose prolotherapy(DPT)in the treatment of postherpetic neuralgia.Methods·Seventy-eight patients with postherpetic neuralgia who visited the Department of Pain of The Affiliated Hospital of Xuzhou Medical University from June 2019 to December 2022 were selected.The patients were randomly assigned to a control group and a research group in a 1∶1 ratio,with 39 patients in each group.The control group was treated with traditional analgesic solution,while the research group was treated with traditional analgesic solution combined with DPT.Visual analog scale(VAS)was used to evaluate the patients'pain level before and after treatment,flow cytometry was used to measure the patients'T-cell subsets,and enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of C-reactive protein(CRP),interleukin-6(IL-6),and IL-10 cytokines.The VAS scores were compared between the two groups of patients before and at 1,2,4,8,and 12 weeks after treatment.CD4+/CD8+,CRP,IL-6,IL-10 levels,and the incidence of adverse reactions before and 2 weeks after treatment were compared between the two groups.Results·There was no statistically significant difference in sex ratio,age,and disease duration between the two groups of patients.The VAS scores of the two groups of patients at 1,2,4,8,and 12 weeks after treatment were significantly lower than those before treatment,and the differences were statistically significant(all P<0.05).The VAS scores of the research group at 1,2,4,8,and 12 weeks after treatment were significantly lower than those of the control group(all P<0.05).There was no statistically significant difference in basal CD4+/CD8+,CRP,IL-6 and IL-10 levels between the two groups of patients.IL-6 and CRP levels in the research group were significantly lower after treatment than those in the control group,and the differences were statistically significant(all P=0.000).CD4+/CD8+ and IL-10 levels were significantly higher in the research group than those in the control group after treatment,and the difference was statistically significant(all P=0.000).No adverse reactions such as local nerve damage,epidural hematoma,infection,pneumothorax or allergy occurred in both groups of patients during the treatment.Conclusion·DPT can significantly reduce the pain of PHN patients,improve patients'T lymphocyte subpopulations and cytokine expression,and can be safely applied to the clinic.

Objective:To observe any effect of early recumbent treadmill training on the balance and functional independence during hospitalization of children who have received hematopoietic stem cell transplantation (HSCT).Methods:This was a retrospective analysis of 106 children who had received HSCT. Sixty-nine of them were qualified for study. Of those, 32 had performed recumbent treadmill training and the other 37 had not. The children in both groups received routine clinical treatment and nursing care, and also health education advocating exercise and giving exercise programs before and after the transplantation. The daily exercise was conducted with the help of parents. It lasted 20 to 30 minutes each time, 4 or 5 times a week. The treadmill group additionally spent 30 minutes training on a recumbent treadmill 5 times a week for 6 weeks. Balance, functional independence and fatigue levels were quantified before and after the treatment using the Berg Balance Scale (BBS), the Functional Independence Measure for Children (WeeFIM) and the Pediatric Quality of Life Inventory-Multidimensional Fatigue Scale.Results:After the 6 weeks, significant improvement was observed in the experimental group′s average BBS score, motor function domain score, total WeeFIM score, general fatigue, and sleep/rest fatigue. All were then significantly better than the non-treadmill group′s results.Conclusion:Early recumbent treadmill training can promote the recovery of balance and functional independence of children after HSCT.

Objective To investigate the infection status and changing trend of hepatitis C virus(HCV)infection in hospitalized patients in medical institutions,and provide reference for formulating HCV infection prevention and control strategies.Methods HCV infection surveillance results from cross-sectional survey data reported to China Healthcare-associated Infection(HAI)Surveillance System in 2020 were summarized and analyzed,HCV positive was serum anti-HCV positive or HCV RNA positive,survey result was compared with the survey results from 2003.Results In 2020,1 071 368 inpatients in 1 573 hospitals were surveyed,738 535 of whom underwent HCV test,4 014 patients were infected with HCV,with a detection rate of 68.93％and a HCV positive rate of 0.54％.The positive rate of HCV in male and female patients were 0.60％and 0.48％,respectively,with a statistically sig-nificant difference(x2=47.18,P＜0.001).The HCV positive rate in the 50-＜60 age group was the highest(0.76％),followed by the 40-＜50 age group(0.71％).Difference among all age groups was statistically signifi-cant(x2=696.74,P＜0.001).In 2003,91 113 inpatients were surveyed.35 145 of whom underwent HCV test,resulting in a detection rate of 38.57％;775 patients were infected with HCV,with a positive rate of 2.21％.In 2020,HCV positive rates in hospitals of different scales were 0.46％-0.63％,with the highest in hospital with bed numbers ranging 600-899.Patients'HCV positive rates in hospitals of different scales was statistically signifi-cant(X2=35.34,P＜0.001).In 2020,12 provinces/municipalities had over 10 000 patients underwent HCV-rela-ted test,and HCV positive rates ranged 0.19％-0.81％,with the highest rate from Hainan Province.HCV posi-tive rates in different departments were 0.06％-0.82％,with the lowest positive rate in the department of pedia-trics and the highest in the department of internal medicine.In 2003 and 2020,HCV positive rates in the depart-ment of infectious diseases were the highest,being 7.95％and 3.48％,respectively.Followed by departments of orthopedics(7.72％),gastroenterology(3.77％),nephrology(3.57％)and general intensive care unit(ICU,3.10％)in 2003,as well as departments of gastroenterology(1.35％),nephrology(1.18％),endocrinology(0.91％),and general intensive care unit(ICU,0.79％)in 2020.Conclusion Compared with 2003,HCV positive rate decreased significantly in 2020.HCV infected patients were mainly from the department of infectious diseases,followed by departments of gastroenterology,nephrology and general ICU.HCV infection positive rate varies with gender,age,and region.

The incidence of intrahepatic cholangiocarcinoma (ICC) continues to rise, and there are no effective drugs to treat it. The immune microenvironment plays an important role in the development of ICC and is currently a research hotspot. Icaritin (ICA) is an innovative traditional Chinese medicine for the treatment of advanced hepatocellular carcinoma. It is considered to have potential immunoregulatory and anti-tumor effects, which is potentially consistent with the understanding of "Fuzheng" in the treatment of tumor in traditional Chinese medicine. However, whether ICA can be used to treat ICC has not been reported. Therefore, in this study, sgp19/kRas, an in situ ICC mouse model with intact immune system, was selected to evaluate the efficacy of ICA in the treatment of ICC in vivo for the first time, and the effects of ICA on the tumor immune microenvironment of ICC mice were analyzed by flow cytometry. This experiment was approved by the Experimental Animal Ethics Committee of Capital Medical University (approval number: AEEI-2023-138). In this study, sgp19/kRas ICC mouse model was treated with oral gavage of 100 mg·kg^-1 ICA. We found that ICA significantly inhibited tumor growth and tumor cell proliferation in the sgp19/kRas ICC mouse model after 3 weeks of treatment. Flow cytometry analysis indicated that ICA treatment markedly reduced the proportion of M2 macrophages and increased the number of CD3⁺CD4⁺ T cells. In vitro experiments demonstrated that ICA promoted macrophage polarization towards the M1 phenotype while inhibiting polarization towards the M2 phenotype. Furthermore, transcriptomic analysis suggested that ICA enhanced Toll-like receptor 9 (TLR9) expression, influencing macrophage nitric oxide metabolism synthesis pathways and receptor-related activities, thereby regulating macrophage polarization. In summary, this study demonstrates that ICA treatment significantly delays tumor progression in sgp19/kRas ICC mouse models. Mechanistically, ICA may achieve its anti-ICC effects by upregulating TLR9 receptor expression levels, promoting macrophage polarization towards the M1 phenotype, and altering the tumor immune microenvironment.

Objective To establish a nude mouse model of type 2 diabetes mellitus(T2DM)and pancreatic cancer that allows dynamic observation of tumor formation process and facilitates in vivo research.Methods At first,human pancreatic cancer PANC-1 cells were transfected with lentiviral vector GV260 to construct the pancreatic cancer cell line PANC-1-Luc with stable expression of firefly luciferase.Then,36 specific pathogen-free nude mice were randomly divided into control group with 12 mice and model group with 24 mice(nude mice with T2DM and pancreatic cancer).The mice in the control group were fed with breeding diet and were then given ectopic subcutaneous implantation of PANC-1-Luc cells,and those in the model group were first given high-fat diet and intraperitoneal injection of 1%STZ,followed by ectopic subcutaneous implantation of PANC-1-Luc cells.The fluorescence in vivo imaging system and the manual measurement method were used for simultaneous and dynamic monitoring of the growth of pancreatic cancer in nude mice in the two groups,and the tumor growth curve was plotted to investigate the correlation between fluorescence value and tumor volume.Subcutaneous tumors and pancreatic islets were observed under a microscope to verify whether the model was successfully established,and immunohistochemistry was used to measure the expression of Ki-67 in tumor tissue to investigate the influence of hyperglycemia on the growth of pancreatic cancer in nude mice.The independent-samples t test was used for comparison of normally distributed continuous data between groups,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups.Results The optimal virus titer was determined as 5×107 TU/mL for the stable transfection of lentiviral vector in PANC-1 cells,and the optimal concentration selected with puromycin was 20 μg/mL,with an optimal selection time of 9 days.The fluorescence value of PANC-1-Luc cells was linearly and positively correlated with the number of cells,with the linear equation of y=42.56x-42 504(r=0.977,P=0.004).The blood glucose value of T2DM nude mice was 23.05(19.25-26.40)mmol/L,with a blood glucose level of＞11.1 mmol/L in each nude mouse,and there was a significant difference in blood glucose value between the T2DM nude mice and the control nude[6.15(5.20-7.30)mmol/L](Z=-8.45,P＜0.001).Compared with the control group,the model group had reductions in the number and volume of pancreatic islets,with irregular shapes and unclear boundaries,and pathological examination confirmed that the xenograft tumor was pancreatic cancer tissue,which showed that the model was established successfully.In the model group,there was a linear positive correlation between subcutaneous tumor size and fluorescence values,with the linear equation of y=232 348 691x-8 258 608(r=0.911,P=0.031).The model group had a significantly higher positive rate of Ki-67 than the control group(50.333%±7.808%vs 15.917%±4.055%,t=13.55,P＜0.001),suggesting rapid tumor proliferation in the model group.Conclusion The T2DM nude mouse model of pancreatic cancer established in this study can simulate the pathological process of the development and progression of pancreatic cancer in the context of T2DM and dynamically observe the influence of hyperglycemia on the growth of pancreatic cancer cells in vivo,thereby providing a new experimental vector for the in vivo study of the development and progression of pancreatic cancer in the context of T2DM.

Objective To investigate the effect of Anchusa italica Retz.extract on cough variant asthma(CVA)and its mechanism of action.Methods Forty-eight SD rats of each eight were randomly divided into blank group(equal amount of saline),model group(equal amount of saline),control group(250 mg·kg-1 prednisone acetate)and experimental-A1,-A5,-A7 groups(61.1 mg·kg-1 Anchusa italica Retz.extract A1,48.8 mg·kg-1 Anchusa italica Retz.extract A5,201.8 mg·kg-1 Anchusa italica Retz.extract A7),continuous gavage for 28 d;except for the blank group,the other groups were made with ovalbumin-complete Freund's adjuvant.The number of cough times of each rat was observed.The changes in the expression of Toll-like receptor 4(TLR4),myeloid differentiation factor(MyD88)and nuclear factor κB(NF-κB)proteins were observed in the lung tissue.Results The cough times in the blank group,model group,control group and experimental-A1,-A5,-A7 groups were 1.10±0.22,8.33±1.24,3.08±0.65,3.31±0.99,3.08±1.02 and 3.06±0.68;the relative expression levels of TLR4 protein were 0.61±0.01,0.84±0.04,0.66±0.02,0.64±0.04,0.64±0.03 and 0.69±0.02;the relative expression levels of MyD88 protein were 0.54±0.08,0.86±0.06,0.71±0.06,0.65±0.05,0.64±0.08 and 0.70±0.06;the relative expression levels of p-NF-κB protein were 0.48±0.11,0.94±0.06,0.80±0.08,0.68±0.04,0.68±0.06 and 0.78±0.09.Compared with the experimental-A1,-A5,-A7 groups,control group,normal group,the differences were statistically significant in the model group(all P＜0.05).Conclusion The extract of Anchusa Italica Retz.can inhibit the symptoms of CVA rats,and the mechanism may be related to the inhibition of TLR4/MyD88/NF-κB signaling pathway.

Background::A phase II trial on recombinant human tenecteplase tissue-type plasminogen activator (rhTNK-tPA) has previously shown its preliminary efficacy in ST elevation myocardial infarction (STEMI) patients. This study was designed as a pivotal postmarketing trial to compare its efficacy and safety with rrecombinant human tissue-type plasminogen activator alteplase (rt-PA) in Chinese patients with STEMI.Methods::In this multicenter, randomized, open-label, non-inferiority trial, patients with acute STEMI were randomly assigned (1:1) to receive an intravenous bolus of 16 mg rhTNK-tPA or an intravenous bolus of 8 mg rt-PA followed by an infusion of 42 mg in 90 min. The primary endpoint was recanalization defined by thrombolysis in myocardial infarction (TIMI) flow grade 2 or 3. The secondary endpoint was clinically justified recanalization. Other endpoints included 30-day major adverse cardiovascular and cerebrovascular events (MACCEs) and safety endpoints.Results::From July 2016 to September 2019, 767 eligible patients were randomly assigned to receive rhTNK-tPA ( n = 384) or rt-PA ( n = 383). Among them, 369 patients had coronary angiography data on TIMI flow, and 711 patients had data on clinically justified recanalization. Both used a –15% difference as the non-inferiority efficacy margin. In comparison to rt-PA, both the proportion of patients with TIMI grade 2 or 3 flow (78.3% [148/189] vs. 81.7% [147/180]; differences: –3.4%; 95% confidence interval [CI]: –11.5%, 4.8%) and clinically justified recanalization (85.4% [305/357] vs. 85.9% [304/354]; difference: –0.5%; 95% CI: –5.6%, 4.7%) in the rhTNK-tPA group were non-inferior. The occurrence of 30-day MACCEs (10.2% [39/384] vs. 11.0% [42/383]; hazard ratio: 0.96; 95% CI: 0.61, 1.50) did not differ significantly between groups. No safety outcomes significantly differed between groups. Conclusion::rhTNK-tPA was non-inferior to rt-PA in the effect of improving recanalization of the infarct-related artery, a validated surrogate of clinical outcomes, among Chinese patients with acute STEMI.Trial registration::www.ClinicalTrials.gov (No. NCT02835534).

Objective To investigate the clinical efficacy of standardized comprehensive pure traditional Chinese medicine(TCM)treatment for short-course sudden deafness and to explore the influencing factors of clinical efficacy.Methods The clinical data of 516 short-course sudden deafness inpatients(TCM group)who were given standardized comprehensive pure TCM treatment were retrospectively analyzed,and their efficacy were compared with the efficacy of 1 024 cases in the Chinese multicenter clinical study of sudden deafness(western medicine group).The analysis was carried out for the influence of the gender,left or right ear,age group,degree of deafness,type of hearing curve,concomitant symptoms(tinnitus,vertigo,and dullness of the ear),and the TCM syndrome types on the efficacy.Results(1)The comparison of baseline data showed that in comparison with the western medicine group,TCM group had relatively low proportion of low-frequency descending type while relatively high proportion of total deafness type,and had relatively high treatment difficulty.The course of treatment in TCM group was shortened by nearly 2/3 in comparison with the western medicine group(12.60 days vs 30 days).(2)Among the 516 patients of the TCM group,175 cases were cured,121 cases were markedly effective,90 cases were effective and 130 cases were ineffective,and the total effective rate was 74.80%.Compared with the 1 024 cases in the western medicine group,there was no significant difference in the overall efficacy or the efficacy evaluated with the hearing curve classification(P＞0.05).(3)Analysis of influencing factors of curative effect showed that the type of hearing curve was closely related to the curative effect of sudden deafness(P＜0.001).The curative effect of low frequency descent type was the best,and the curative effect of total deafness type was the worst.The cure of deafness was correlated with the degree of deafness,the milder the degree of deafness,the greater the likelihood of recovery(P＜0.001).The curative effect of the patients without vertigo was superior to that of the patients with vertigo(P＜0.001).The curative effect of adolescent patients was superior to that of middle-aged and elderly patients(P＜0.05).There was no correlation between TCM syndrome types and curative effect(P＞0.05).Conclusion Standardized comprehensive pure TCM treatment exerts certain curative effect on short-course sudden deafness,and may have some advantages compared with conventional western medicine treatment.The type of hearing curve,degree of deafness,accompanied by vertigo or not,and age group are the influencing factors of the efficacy.

Objective To investigate the effects of exercise and complex environment on lipopolysaccharide(LPS)-induced dopaminergic neuron death in the substantia nigra of midbrain.Methods C57BL/6 mice were divided into control group,LPS group,LPS+swimming group and LPS+complex environment group,with 7 mice in each group.The mice in the LPS group were injected with LPS into the brain to establish an inflammatory model of Parkinson's disease and lived in cages for 2 weeks.Mice in LPS+swimming group were forced to swim for 15 minutes every day for 2 weeks after modeling.The mice in the LPS+complex environment group were placed in a complex environment for 2 weeks after modeling.The control group mice were not treated.After 14 days of modeling,behavioral experiments such as footprint,open field and rotating rod were performed on each group of mice to detect the autonomous exercise ability,exercise balance ability and depression level of mice.The expressions of tyrosine hydroxylase(TH)in substantia nigra was detected by immunohistochemical staining and Western blotting.The expressions of brain-derived neurotrophic factor(BDNF),Caspase-3,interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the substantia nigra of the midbrain were detected by Western blotting.The transcription levels of IL-1β,IL-6 and TNF-α in substantia nigra were detected by RT-PCR.Results Compared with the control group,the exercise ability and balance ability of mice in LPS group,LPS+swimming group and LPS+complex environment group decreased,the depression level increased(P＜0.001),the number of TH positive neurons and BDNF protein decreased significantly(P＜0.001),and the contents of Caspase-3,IL-1β,IL-6 and TNF-α increased significantly(P＜0.001).Compared with the LPS group,the exercise ability and balance ability of the mice in the LPS+swimming group and the LPS+complex environment group were restored,the depression level decreased significantly(P＜0.01),the survival number of TH positive neurons and the content of BDNF increased significantly(P＜0.01),Caspase-3,IL-1β,IL-6 and TNF-α reduced significantly(P＜0.01),and the phenomenon in the LPS+complex environment group was more significant.Conclusion Exercise and complex environment can inhibit LPS-induced central nervous system inflammation in mice,thereby reducing damage to midbrain substantia nigra neurons,and the inhibitory effect of LPS+complex environment group is more significant.