Objective To explore the changes in Th1 cell,B cell and related gene expression during the healing of diabetic foot ulcers(DFU).Methods Lesional skin tissues from DFU patients surgically treated in our hospital from January 2022 to January 2023 were selected.DFU associated gene expression data were collected from GSE80178 and GSE143735 datasets.The expressions of genes(DEGs)between ulcer and no-ulcer patients were identified.Bioinformatics methods were used to identify abnormal immune cells and key genes.Finally,the key results were detected by RT-qPCR and flow cytometry.Results A total of 548 common DEGs were identified in two datasets.In 14 co-expression modules,darkgrey and darkgreen were most related to ulcer,which were mainly associated with the regulation of enzyme activity,immune function and endocrine regulation.Flow cytometry showed that Th1 and B cells were highly infiltrated in ulcer tissue and decreased in healing tissue.MMP13,S100A9 and STAT4 were involved in immune signaling pathways.MMP13 and STAT4 were lowly expressed in healed ulcers,whereas S100A9 was highly expressed.Conclusion Reduced levels of Th1 and B cell infiltration may promote DFU healing.

Objective: As a gate-keeper enzyme link, pyruvate dehydrogenase E1 subunit alpha (PDHA1) functions as a key regulator during glycolysis and the mitochondrial citric acid cycle, which has been reported in several tumors. Nevertheless, the effects of PDHA1 on biological behaviors and metabolism remain unclear in cervical cancer (CC) cells. The study aims to explore the PDHA1 effects on glucose metabolism in CC cells and its possible mechanism. Methods: We first determined the expression levels of PDHA1 and activating protein 2 alpha (AP2α) as a PDHA1 potential transcription factor. The effects of PDHA1 in vivo were evaluated through a subcutaneous xenograft mouse model. Cell Counting Kit-8 assay, 5-ethynyl-2′-deoxyuridine (EdU) labeling assay, Transwell invasion assay, wound healing assay, Terminal deoxynucleotidyl transferase dUTP nick end labeling assay and flow cytometry were performed in CC cells. Oxygen consumption rate (OCR) levels were determined to reflect aerobic glycolysis level in gastric cancer cells. Reactive oxygen species (ROS) level was measured with 2′, 7′-dichlorofluorescein diacetate kit. The relationship between PDHA1 and AP2α was examined by conducting chromatin immunoprecipitation assay and electrophoretic mobility shift assay. Results: In CC tissues and cell lines, PDHA1 was downregulated, while AP2α was upregulated. Overexpression of PDHA1 remarkedly inhibited the proliferation, invasion and migration of CC cells, and tumor growth in vivo, as well as promoted OCR, apoptosis and ROS production. Moreover, AP2α directly bound to PDHA1 within suppressor of cytokine signaling 3 promoter region to negatively regulate PDHA1 expression level. What is more, PDHA1 knockdown could effectively reversed the AP2α silencing-mediated suppressive effects on cell proliferation, invasion, migration, and the promotive effects of AP2α knockdown on OCR, apoptosis and ROS production. Conclusions Our findings demonstrate that AP2α negatively regulated PDHA1 via binding to PDHA1 gene promoter to promote malignant CC cell behaviors, which may provide a potential approach for CC therapeutics.

This article summarizes the whole-person care needs of patients in Intensive Care Unit (ICU) from the physiological, psychosocial, and spiritual aspects. This article reviews the whole-person care model of ICU palliative care from three aspects, namely physiological care, psychosocial support and spiritual support, so as to provide a reference for the development of ICU palliative care in China.

Objective To compare the efficacy and safety of the long-term intermittent maintenance treatment with tacrolimus 0.03％ ointment versus traditional treatment in reducing relapses and prolonging the recurrence interval in children with moderate to severe atopic dermatitis (AD).Methods A two-phase randomized,open-labelled,controlled clinical trial was conducted from September 2012 to November 2013.In the first phase,a total of 171 children aged 2-15 years with moderate to severe AD were enrolled from 7 hospitals in China,and received conventional treatment with tacrolimus 0.03％ ointment twice a day for 2-6 weeks.At the end of the treatment,the patients who achieved an investigator's global assessment (IGA) score ≤ 2 (n =125) were randomly classified into 2 groups to receive the second-phase treatment:test group (n =62) receiving intermittent maintenance treatment with tacrolimus 0.03％ ointment twice a week (Monday and Thursday),and control group (n =63) receiving no treatment.If the patients in the 2 groups experienced relapse,they received conventional treatment with tacrolimus 0.03％ ointment twice a day.The overall observation period was 6 months.The primary endpoint was the time to the first relapse,which was defined as the number of days from the end of the first-phase treatment to the first relapse.The secondary endpoints included the number of relapses at the second-phase trial,the disease severity at the time of relapse,the duration of relapse,the pruritus score at the time of relapse,the total amount of tacrolimus ointment used,the total response rate at the second-phase trial,and the incidence of adverse events.Results A total of 125 children with AD were enrolled into the second-phase trial,and 121 of them completed the follow-up.Among the 121 patients,the recurrence rate was significantly lower in the test group (25/60,41.7％) than in the control group (46/61,75.4％;x2 =14.20,P ＜ 0.001).The time to the first relapse was significantly longer in the test group (46.9 ± 37.7 d) than in the control group (28.8 ± 32.3 d;Z =1 093.50,P =0.020).The total number of recurrence was 31 and 86 in the test group and control group respectively,and the mean number of recurrence in each patient was significantly lower in the test group (0.52 ± 0.68) than in the control group (1.41 ± 1.23,t =4.96,P ＜ 0.001).There were no significant differences between the two groups regarding disease severity during relapse (eczema area and severity index:Z =971.50,P =0.39),duration of relapse (Z =747.00,P =0.07),and pruritus score during relapse (Z =894.00,P =0.95).The therapeutic drug was tolerated well in all the children,and no tacrolimus-related serious adverse events occurred.Conclusion The intermittent maintenance treatment with tacrolimus 0.03％ ointment twice a week for 6 months can effectively and safely prevent and reduce relapses,and prolong the recurrence interval in children with moderate to severe AD.

Objective To explore the association between post-stroke blood pressure and collaterals and functional outcome in acute ischemic stroke (AIS) patients with large vessel occlusion/stenosis.Methods Consecutive AIS patients with large vessel occlusion/stenosis who had symptom onset within 6 hours in Huashan Hospital from July 2011 to October 2017 were retrospectively recruited and enrolled.Collaterals were analyzed through CT perfusion imaging.And all the patients were followed up for 3 months.And the association between baseline blood pressure,collaterals and outcomes was analyzed using multivariate linear and Logistic models.A total of 126 patients were enrolled for multivariate Logistic and linear regression analysis.Results Multivariate Logistic regression analysis revealed that with every increment of 10 mmHg (1 mmHg =0.133 kPa) systolic blood pressure,the odds of poor functional outcome increased by 20％ (unadjusted OR =1.20,95％ CI 1.02-1.42,P =0.03).After adjusting for age and sex,the odds increased by 21％ (OR =1.21,95％ CI 1.01-1.46,P =0.04).However,no statistical significance was shown after multivariate adjustment (OR--1.02,95％ CI 0.99-1.05,P =0.06).Multivariate linear regression analysis demonstrated that diastolic blood pressure (DBP) was negatively associated with the volume of hypoperfused and severely hypoperfused lesion.With every increment of 10 mmHg DBP,the volume of delay time (DT) ＞ 6 s lesion downsized by 7.6 ml (unstandardized coefficient -7.56,95％ CI-14.89--0.24,P =0.04).The DT ＞6 s/DT ＞3 s ratio was also decreased with the elevation of DBP (unstandardized coefficient-0.03,95 ％ CI-0.06--0.01,P =0.01).Conclusions For AIS patients,though higher DBP seems to be associated with better collaterals,it may still relate with poor functional outcome.

Objective To explore the relation of gamma knife radiation doses and changes of trigeminal nerves after gamma knife irradiation by studying the changes of trigeminal somatosensory evoked potentials (TSEP) characteristics oftrigeminal nerve roots irradiated by gamma knife in normal animals.Methods Fifteen New Zealand rabbits were divided into 60 Gy irradiation group,80 Gy irradiation group,100 Gy irradiation group,120 Gy irradiation group and control group in accordance with random number table (n=3).The first 4 groups used Gamma Plan 5.34 planning system to design the radiation targets and radiation doses.The target was placed at trigeminal nerve root entry zone,bilateral irradiation was performed,and the central dose was divided into 60 Gy,80 Gy,100 Gy and 120 Gy.The bilateral TSEP was monitored before and 6 months after irradiation.The monitoring data were recorded and statistical analysis was performed.Results P1,N1,P2 wave latencies,P1-N1,N1-P2 interpeak latencies and amplitudes in the 60 Gy irradiation group after irradiation showed no significant differences as compared with those in the control group (P＞0.05);P1,N1,P2 wave latencies,P1-N1,N1-P2 interpeak latencies and amplitudes in the 80 Gy irradiation group,100 Gy irradiation group,and 120 Gy irradiation group after irradiation showed significant differences as compared with those in the control group (P＜0.05),and these changes increase with increase of doses.Conclusion The changes of TSEP,which increase with increase of doses,are closely related to the doses of gamma knife irradiation,of which 80 Gy is the lowest dose to produce changes.

Objective To explore the effect of apolipoprotein E (ApoE) on matrix metalloproteinases-9 (MMP-9) expression and its mechanism in astrocytes.Methods (1) Astrocytes in ApoE gene knock out (ApoE-/-) and wild type (WT) C57BL/6J suckling mice were cultured in vitro;glial fibrillary acidic protein antibodies were employed to identify the astrocytes;MMP-9.antibodies were employed to detect the MMP-9 expression in the astrocytes.(2) Astrocytes from the ApoE-/-and wild type WT C57BL/6J suckling mice were divided into activation group,antibody groups and control group randomly;cells in the control group did not give any treatment;cells in the activation group were given whooping cough toxin and thermal inactivation-mycobacterium tuberculosis H37Ra;cells in the antibody groups were given anti-interleukin-6 (IL-6),interferon gamma (IFN--γ),tumor necrosis factor-α (TNF-α) and IL-12,respectively,to inhibit their inflammatory factors.ELISA was performed to detect the concentrations of MMP-9 and tissue inhibitor of metalloproteinase-1 (TIMP-1),and the inflammatory factors concentrations ofTNF-,IL-12,IL-6,and IFN-γ.Results These two kinds of rat astrocytes could both qualitatively express MMP-9;as compared with the control group,the activation group had significantly higher MMP-9 concentration (P＜0.05);the activation group had significantly higher MMP-9 concentration as compared with the antibody groups (P＜0.05);activation group from ApoE-/-C57BL/6J suckling mice had significantly higher MMP-9 concentration than that from wild type C57BL/6J suckling mice (P＜0.05).The concentration of TIMP-1 was not significantly different among various groups (P＞0.05).The concentrations of inflammatory factors in the activation groups from two kinds of mice were significantly higher than those in the control group (P＜0.05);the concentrations of IL-6,IFN-γ TNF-α and IL-12 in each antibody group were significantly lower than those in the activated group (P＜0.05);the concentrations of inflammatory factors in ApoE-/-rats were significantly higher than those in WT rats (P＜0.05).Conclusion ApoE can regulate the MMP-9 expression by regulating the secretion of proinflammatory cytokines,which can affect the integrity of blood brain barrier.

Objective To explore the relation of gamma knife radiation of different doses with trigeminal nerve radiation damage by investigating the pathologic changes of trigeminal nerve roots after different doses of gamma knife radiation.Methods Fifteen New Zealand rabbits were randomly divided into 60 Gy radiation group,80 Gy radiation group,100 Gy radiation group,120 Gy radiation group and control group (n=3).Bilateral trigeminal nerves of all New Zealand rabbits,excepted for the control group,were irradiated;and a 4-mm isocenter was placed at trigeminal nerver root entry zone.All New Zealand rabbits for pathological observation were sacrificed after six months.The roots were examined by light microscopy,immunohistochemistry,and electron microscopy.Integrated optical density of the myelin staining and immunohistochemistry staining results of the four groups were examined by Image-pro plus 6.0 image processing system,and analyzed using method of statistics.Results Pathological observation showed the result of 60 Gy group after irradiation did not differ significantly as compared with the result of control group;in 80 Gy group,there was part nerve fiber fragmentation and degeneration,and demyelination;in 100 Gy group,there was nerve fiber fragmentation,degeneration and demyelination,and part of nervous necrosis;in 120 Gy group,there was almost all nerve fiberdissolution,disappearance and demyelination,and most of nervous necrosis.The integrated optical density of the myelin staining and immunohistochemistry staining results of 60 Gy group did not differ significantly as compared with that of the control group (P＞0.05);the results of 80 Gy,100 Gy and 120 Gy group differed significantly as compared with that of control group (P＜0.05) and decreased following the increase of irradiation doses.Conclusions The histopathological changes of the trigeminal nerve irradiated by gamma knife have significant correlation with irradiation doses.The result of 60 Gy group after irradiation does not change significantly;the radiation injury of 80 Gy-120 Gy group aggravates with the increase of irradiation doses;irradiation of high dose (100 Gy and 120 Gy) can cause nervous necrosis.

Objective To observe recombinant plasmids were constructed with the macrophage colony-stimulating factor ( GM-SCF) , interleukin -21 (IL-21) and retinoic acid early transcription factor-1 (Rae-1), and observe the inhibitory effects in subcutaneous liver cancer model in mice with the recombinant plasmids.Methods The recombinant plasmids of GM-SCF, IL-21 and Rae-1 were constructed with RT-PCR method, mouse model was constructed, the model mice were randomly divided into six groups including control, IRES/GFP, IRES/IL21, IRES/GM-SCF, IRES/GM-SCF-IL21 and IRES/combination with 10 mice included in each group, each groups (15 mice) were treated with the corresponding gene therapy.The survival rate were observed after 60 days.The blood levels of interferon -γ(IFN-γ) and interleukin -2 (IL-2) were detected in each group.Results The pGM-CSF-GFP-IRES-Rae-1-IL-21 has been successfully constructed.All mice had demised 14 and 16 days after treatment in the control and IRES/GFP groups, respectively.There were 2, 1, 11 mice remaining after 60 days of treatment in the IRES/GM-SCF, IRES/IL21 and IRES/GM-SCF-IL21 groups respectively.The survival rate of mice at 60 days of treatment was 73.33%, 13.33%, and 6.67% for groups IRES/GM-SCF-IL21, IRES/GM-SCF and IRES/IL21, respectively.The survival rate of the mice was significantly higher in IRES/GM-SCF-IL21 than the other groups.The levels of IL-2 and INF-γof mice 1-6 days after treatment gradually increased in the IRES/combination groups, including IRES/GM-SCF-IL21, IRES/GM-SCF and IRES/IL21.They were highest in the IRES/combination group and lowest (P<0.01) in the IRES/GM-SCF and IRES/IL21 groups, with the IRES/GM-SCF-IL21 group showing intermediate levels.By 6-10 days after treatment, IL-2 and INF-γlevels had stably increased in the IRES/combination groups, but had gradually decreased in the IRES/GM-SCF-IL21, IRES/GM-SCF and IRES/IL21 groups.At the end of treatment, IL-2 and INF-γlevels were significantly (P<0.01) higher in the IRES/GM-SCF-IL21 than were found in either the IRES/GM-SCF group or IRES/IL21 group, which were also significantly (P<0.01) higher than either the IRES/GFP or control groups.The levels of IL-2 and INF-γwere highest in the IRES/combination group ( P<0.01) and not significantly different among the IRES/GM-SCF, IRES/IL21, IRES/GFP, and control groups.Conclusion The inhibitory effects in subcutaneous liver cancer model in mice were obvious significantly, and its mechanism maybe be related to the activation of the body's immune.

Objective To investigate the prevalence and progression process of atopic diseases in adolescents, and to assess their relationship with filaggrin(FLG)mutations. Methods Totally, 334 adolescents aged from 11 to 19 years in a middle school in shanghai were enrolled into this study. A clinical interview was carried out to determine the prevalence of atopic diseases (such as ichthyosis, atopic dermatitis (AD), asthma, rhinitis, etc)in these subjects. Peripheral blood samples were collected from 285 out of the 334 adolescents for screening for common FLG mutations, including 3321delA and K4671X. Five years later, these adolescents were followed up for reevaluation of clinical presentations of atopic diseases. Statistical analysis was carried out by the chi-square test with the SPSS 20.0 software. Results As the baseline survey showed, 19 (5.69%)of the 334 adolescents had AD, 14 (4.19%)had ichthyosis vulgaris, 36(10.78%)had allergic rhinitis, and 4(1.20%)had asthma. FLG mutations were observed in 24(8.42%) of the 285 adolescents. Five years later, 265 adoscents completed the follow-up, and 69 (20.66%)were lost to follow-up. Of the 265 adolescents reevaluated, 13(4.89%)had AD, 15(5.64%)had ichthyosis vulgaris, 27(10.15%)had allergic rhinitis, and 1 (0.38%)had asthma. By the time the second survey was performed, 6 out of the 19 patients initially diagnosed with AD had achieved complete regression, 13 had experienced a marked decrease in SCORing atopic dermatitis (SCORAD)score, and symptoms had disappeared in 9 of the 36 patients initially diagnosed with allergic rhinitis. The frequency of FLG mutations was 10.0%in patients with AD, 55.6%in those with ichthyosis, and 40.0%in those with both AD and ichthyosis, and the development of ichthyosis was associated with FLG mutations(P<0.001). Conclusions The frequency of common FLG mutations was 8.42%in these adolescents. FLG gene may be a semidominant gene associated with ichthyosis vulgaris, and multiple factors influence its expression.