Objective To search for new biomarkers to predict prognosis in colorectal cancer (CRC) patients.Methods A prognostic model was developed for colorectal cancer with immune-related genes from the cancer ge-nome atlas (TCGA) database using one-way Cox regression analysis and least absolute shrinkage and selection op-erator (LASSO) regression analysis.Moreover, the immune infiltration characteristics of patients in high and low risk groups was compared by sstimation of stromal and immune cells in malignant tumor tissues using expression da-ta (ESTIMATE) and cell-type identification by estimating relative subsets of RNA transcripts (CIBERSORT) .In addition, the expression levels of immune checkpoints were analyzed in patients from different risk groups.The sen-sitivity of patients in the two risk groups to chemotherapeutic agents was also compared based on genomics of drug sensitivity in cancer (GDSC).Results It was found that the prognostic model constructed based on immune genes could better predict the overall survival (OS) of CRC patients,and the results showed area under curve (AUC) values of 0.764 (95％ CI:0.751-0.793), 0.773 (95％ CI:0.761 -0.779), and 0.760 (95％ CI:0.742 -0.774) for 1-, 3-, and 5-year OS, respectively.Patients in the low-risk group had higher expression levels of im-mune checkpoints and more abundant immune cells such as T cells (P<0.001) , dendritic cells (P<0.001) , macrophages (P<0.001) , neutrophils (P<0.001) .Patients in the high-risk group might be more sensitive to some chemotherapeutic agents such as axitinib, imatinib, methotrexate, pazopanib, rapamycin, sunitinib and tasig-arnib.Conclusion A prognostic model based on 19 immune genes was effective in predicting the prognosis of CRC patients.The number and activity of immune cells in the immune microenvironment in different patients may be an important factor influencing their response to immunocheck inhibitors and chemotherapeutic agents.

Objective:To analyze the clinical features of MDA5 antibody positive dermatomyositis (MDA5-DM) and to provide evidence for early diagnosis and treatment.Methods:From March 2019 to June 2021, 272 patients with anti-MDA5-DM from the Nanjing Medical University myositis-associated interstitial lung disease cohort were enrolled, with 76 patients with anti-synthetase syndrome (ASS) as the control group. The clinical characteristics and the occurrence of interstitial lung disease were analyzed. T-test was used for normally distributed and variance-homogeneous independent samples, Mann-Whitney U test for non-normally distributed data, and chi-square test or Fisher′s exact test for dichotomous variables. Results:Among the 272 anti-MDA5-DM patients, 88.6% (241/272) developed interstitial lung disease (ILD), and 33.8% (92/272) developed rapidly progressive ILD (RP-ILD). The six-month all-cause mortality rate of anti-MDA5-DM patients was 16.9% (46/272), and it was as high as 47.8% (44/92) for those with RP-ILD. Compared with ASS patients, anti-MDA5-DM patients had a significantly higher proportion of males, arthritis, Gottron's sign, heliotrope rash, V-sign, periungual erythema, and skin ulcers ( P<0.05). The levels of ALT, AST, and ferritin were significantly increased ( P<0.05). Compared with non-RP-ILD patients, RP-ILD patients had a significantly higher proportion of males [35.9%(33/92) vs. 23.3%(42/180), χ2=4.79, P=0.029], higher levels of LDH [387 (276, 547) U/L vs. 310 (245, 400) U/L, Z=-3.67, P<0.001], ESR [45.5 (29.25, 63.25) mm/1 h vs. 31.2 (20, 51) mm/1 h, Z=-3.71, P<0.001], CRP [10.9 (4.1, 25.2) mg/L vs. 4.54 (2.58, 9.08) mg/L, Z=-4.97, P<0.001], ferritin [1 340 (650, 2 000) ng/ml vs. 556 (203, 1 186) ng/ml, Z=-4.40, P<0.001], and a higher proportion of anti-Ro52 antibody and anti-MDA5 antibody co-positivity [87.0%(80/92) vs. 52.2%(94/180), χ2=31.87, P<0.001]. Conclusion:Anti-MDA5-DM patients are prone to develop RP-ILD and have poor prognosis.

Objective:To investigate the role of dendritic cells (DC) in Chlamydia muridarum ( Cm) respiratory infection and their effect on adaptive immune response. Methods:C57BL/6 mice were exposed to 1×10 3 inclusion-forming units (IFU) of Cm through inhalation to establish the mouse model of Cm respiratory infection. The proportion of CD11c + MHCⅡ + DC and the expression of costimulatory molecules (CD40, CD80 and CD86) in spleen tissues were detected by flow cytometry on 0, 3 and 7 d after infection. The expression of IL-12p40, IL-10 and IL-6 at mRNA level in spleen tissues was detected by qPCR. Mouse splenic DC isolated on 7 d after Cm infection were sorted by magnetic beads and then transferred to recipient mice. Th1 response in the recipient mice was measured using intracellular cytokine staining 14 d after infection. Results:Cm respiratory infection induced massive infiltration of DC and promoted the expression of costimulatory molecules on splenic DC. The expression of IL-12 and IL-10 at mRNA level in splenic DC reached the peak on 3 d after infection. Transferring the splenic DC of Cm-infected mice into the recipient mice could alleviate the disease condition in the recipient mice after Cm infection with reduced Cm inclusion-forming units in lung tissues and significantly increased proportion of Th1 cells in lung and spleen tissues. Conclusions:Cm respiratory infection could induce the maturation and activation of DC, which promoted Th1 immune response. DC played an important role in Cm infection.

Objective:To investigate the role of IL-21/IL-21R-mediated CD4 + T cells in Chlamydia muridarum ( Cm) respiratory infection. Methods:C57BL/6 mice (WT mice) and IL-21R -/- mice were used to establish the models of Cm respiratory infection through intranasal inhalation of Cm. Flow cytometry was used to detect the proportion, number, activity and function of CD4 + T cells in lung and spleen tissues at 0, 3, 7 and 14 d after Cm respiratory tract infection. IFN-γ and IL-4 levels in spleen cell culture supernatants were detected by ELISA. Na?ve WT mice were transferred with CD4 + T cells in the spleen tissues of IL-21R -/- mice or WT mice on 7 d after infection and given Cm intranasally 2 h later. Then the mice were weighed daily and sacrificed on 14 d after infection. The bacterial load and pathological changes in lung were analyzed. Flow cytometry was performed to detect the proportions and numbers of neutrophils (CD45 + CD11b + Gr-1 high) and alveolar macrophages (CD45 + F4/80 + CD11c high)as well as the proportions of Th1 (IFN-γ + CD4 + ) and Th2 (IL-4 + CD4 + ) cells. ELISA was also performed to measure IFN-γ and IL-4 levels in spleen cell culture supernatants. Results:Compared with WT mice, IL-21R -/- mice showed elevated numbers and enhanced activation of CD4 + T cells, increased proportion of Th1 cells and decreased proportion of Th2 cells in spleen and lung tissues after Cm respiratory infection. Besides, IFN-γ levels increased, while IL-4 levels decreased in spleen cell culture supernatants of IL-21R -/- mice. After Cm infection, the na?ve WT transferred with CD4 + T cells from IL-21R -/- mice showed less body weight loss, reduced bacterial load and alleviated pathological changes in lung tissues, increased proportion of Th1 cells in lung tissue and higher IFN-γ level in spleen cell culture supernatants. Conclusions:IL-21/IL-21R-mediated CD4 + T cells could aggravate Cm respiratory infection by suppressing Th1 cell immune responses.

Objective:To investigate the role of HIS (hyperinflammatory syndrome) score in predicting the prognosis of anti-melanoma differentiation associated gene 5(MDA5) antibody-positive dermatomyositis (DM) patients with interstitial lung disease (ILD).Methods:A total of 43 patients with anti-MDA5 antibody-positive dermatomyositis and 228 connective tissue disease (CTD) patients with ILD hospitalized in the First Affiliated Hospital of Nanjing Medical University from January 2018 to April 2021 were enrolled into this study. All patients were complicated with ILD and their HIS score were assessed. Non-parametric Mann-Whitney U test, Chi-squared test, Fisher exact probability and receiver operating characteristic (ROC) curve were used for data analysis. Results:The HIS score of 43 patients with anti-MDA5 antibody-positive dermat-omyositis were collected. Among 228 CTD-ILD patients in the control groups, the primary disease consisted of 33(14.5%) anti-synthetase antibody syndrome (ASS), 44(19.3%) rheumatoid arthritis (RA), 65(28.5%) Sj?gren's syndromes (SS), 43 (18.9%) systemic sclerosis (SSc) and 43 (18.9%) systemic lupus erythematosus(SLE). The HIS score of anti-MDA5-positive DM-ILD patients [2(1, 3)] was higher than those in ASS patients [1(0, 2), Z=-2.06, P<0.05] and significantly higher than those in RA-ILD [1(0, 2), Z=-2.87, P<0.01], SS-ILD [0(0,1), Z=-5.78, P<0.01], SSC-ILD [1(0, 1), Z=-3.84, P<0.01] and SLE-ILD [1(0, 2), Z=-3.81, P<0.01]. Comparing HIS score of anti-MDA5-positive DM-ILD patients, the 3-months mortality rate in the low, medium and high groups was 0(0/15), 38.1%(8/21) and 85.7%(6/7). The area under ROC curve (95% CI) was 0.857[(0.747, 0.967), P<0.001] for HIS score in predicting 3-months mortality probability of anti-MDA5-positive DM-ILD. Conclusion:HIS score of anti-MDA5-positive DM-ILD is higher than that in CTD-ILD patients and the baseline level is related to the 3-months mortality rate.

Objective:To investigate the effects of Chlamydia muridarum ( Cm) respiratory tract infection on the infiltration and polarization of alveolar macrophages (AMs) and pulmonary interstitial macrophages (IMs). Methods:A C57BL/6 mouse model of Cm respiratory tract infection was established through nasal inhalation. Flow cytometry was used to detect AMs (CD45 + F4/80 + CD11c + ) and IMs (CD45 + F4/80 + CD11c -) in lung tissues at 0, 3, 7 and 14 d after Cm respiratory tract infection. The proportions of M1 (CD80 + , CD86 + , MHCⅡ + , iNOS + ) and M2 (CD206 + , Arg1 + ) macrophages in AMs and IMs were also detected. Results:(1) Cm respiratory tract infection induced the infiltration of AMs and IMs. Compared with the uninfected group (0 d), the proportions and the numbers of AMs and IMs of were significantly increased 3 d after infection ( P<0.05, P<0.01). The numbers of AMs and IMs reached the peak 7 d after infection ( P<0.001). (2) Compared with the uninfected group, the proportions of CD80 + and CD86 + cells in AMs were significantly up-regulated 3 d after infection ( P<0.05, P<0.01); the proportion of MHCⅡ + cells in AMs increased after infection and reached the peak at 14 d ( P<0.05), while the proportion of CD206 + cells decreased after infection ( P<0.05). (3) Compared with the uninfected group, the proportions of CD80 + and CD86 + cells in IMs were increased 3 d after infection ( P<0.05, P<0.001) and the proportion of MHCⅡ + cells was significantly increased 14 d after infection ( P<0.01), while there was no significant change in the proportion of CD206 + cells. (4) In AMs, the proportion of iNOS + cells increased continuously after infection ( P<0.01), while the proportion of Arg1 + cells decreased continuously after infection, especially at 7 d and 14 d ( P<0.05). In IMs, the proportion of iNOS + cells reached the peak at 7 d ( P<0.001), but the proportion of Arg1 + cells showed no significant change after infection. Conclusions:Cm respiratory tract infection induced the infiltration of AMs and IMs, stimulated the polarization of AMs and IMs towards the M1 phenotype and weakened the polarization of AMs to M2 macrophages, but had no significant influence on the polarization of IMs towards the M2 phenotype.

Objective:To evaluate the clinical efficacy of modified Qiwei Baizhu Powder combined with conventional therapy in the treatment of type 2 diabetes mellitus (T2DM) with abnormal lipid metabolism.Methods:A total of 96 patients with T2DM and abnormal lipid metabolism from March 2018 to March 2021 in Anhui Integrated Traditional Chinese and Western Medicine Hospital who met the inclusion criteria were divided into 2 groups according to the random number table method, with 48 in each group. The control group was treated with conventional western medicine, while the observation group was treated with modified Qiwei Baizhu Powder and treatment of the control group. Both groups were treated for 3 months. TCM syndrome scores were performed before and after treatment. Fasting blood glucose (FPG) and 2 hPG were detected by glucose oxidase method, HbA1c was detected by HPLC, TC, TG, HDL-C and LDL-C were detected by cholesterol peroxidase method, glycerophosphate oxidase method, direct inhibition method and direct surfactant clearance method. Adverse events during treatment were recorded and clinical efficacy was evaluated.Results:The total effective rate was 93.8% (45/48) in the observation group and 79.2% (38/48) in the control group, with a statistically significant difference between the two groups ( χ2=4.36, P=0.037). After treatment, the scores of dry mouth, fatigue, anorexia, dizziness, tongue, pulse and total scores in the observation group were significantly lower than those in the control group ( t values were 3.58, 3.17, 3.24, 3.59, 3.58, 2.76 and 8.44, respectively, all Ps<0.05); the levels of FPG, 2 hPG and HbA1c in the observation group were significantly lower than those in the control group ( t values were 3.37, 2.05 and 3.73 respectively, all Ps<0.05). After treatment, the levels of TC [(4.30±0.85) mmol/L vs. (4.78±0.94) mmol/L, t=2.62], TG [(3.00±0.37) mmol/L vs. (3.19±0.54) mmol/L, t=2.01], LDL-C [(2.60±0.71) mmol/L vs. (2.95±0.44) mmol/L, t=2.90] were significantly lower than those in the control group ( P<0.05). After treatment, the HDL-C [(2.07±0.63) mmol/L vs. (1.82±0.55) mmol/L, t=2.01] level was significantly higher than that of the control group ( P<0.05). Conclusion:Modified Qiwei Baizhu Powder combined with conventional therapy can improve blood glucose and blood lipid levels in T2DM patients with abnormal lipid metabolism, relieve clinical symptoms and improve curative effect.

Objective:To investigate the efficacy of continuous intravenous injection of hyaluronidase (HAase) combined with urokinase (UK) in the treatment of ischemia of hyaluronic acid (HA) arterial embolized skin flap.Methods:(1)Establish an animal model of superficial abdominal artery embolization with HA: 24 SD rats were used to make a square island flap (2 cm×2 cm, the width of the pedicle is 2 mm) on the left and right sides of the abdominal white line with superficial epigastric artery as feeding artery. The left and right flaps of rats were used as experimental group and control group. After the preparation of the experimental skin flap, HA 10 μl was injected into the superficial abdominal artery, while the self-control flap was only made and the vessels were peeled off without embolization.(2)The rats were randomly divided into A, B, C and D groups by drawing lots with 6 rats in each group, After the successful establishment of embolic animal model of 45 min, HAase(2 000 IU/kg)+ UK(50 000 IU/kg), HAase(2 000 IU/kg), UK(50 000 IU/kg) and normal saline were continuously infused through caudal vein with microinjection pump. The volume of solution in each group was 6 ml in 12 minutes. The general condition of rats and the skin color, edema, congestion and skin necrosis in the operation area of rats were observed, which were photographed and compared immediately, and at 3 days, 5 days and 7 days after embolization. The images were analyzed by Photoshop software, and the percentage of survival area of the flap after operation 7 days, was measured by pixel method. The percentage of survival area was compared by single factor analysis of variance (ANOVA) and head-to-head comparison by LSD- t test. Results:Immediately after operation, all the flaps in the four groups were pale and there was no obvious swelling. All the flaps showed different degrees of swelling within 3 days after operation, and the swelling basically disappeared within 3-5 days after operation, and gradually changed from dark red ecchymosis to dark purple or black. Seven days after operation, the necrotic area flap gradually hardened and its boundary was obvious. The skin flaps of the 4 groups showed different degrees of necrosis. The postoperative reaction of the flap was the slightest in group A, the performance of group B and C was similar, which were both between group A and group D, and there was the heaviest postoperative reaction in group D. At 7 days after operation, the percentage of survival area of flaps in groups A, B, C and D was 90.30%±5.95%, 52.63%±6.90%, 51.14%±5.95% and 7.70%±2.18%, respectively. The percentage of survival area of skin flap in group A was significantly higher than that in groups B, C and D (analysis of ANOVA: P<0.01; LSD- t test: P<0.01). Conclusions:Continuous intravenous infusion of HAase combined with UK can effectively alleviate the flap ischemia caused by HA artery embolism, increase tissue perfusion and increase the survival area of the flap.

Objective:To explore the current status of pediatric nurses ' transfer tendency and its influencing factors, so as to provide a basis for reducing pediatric nurses ' transfer tendency. Methods:This research conducted the cross-sectional survey. From July to August 2020, convenience sampling was used to select pediatric nurses from 3 Class Ⅲ Grade A hospitals in Henan Province as the research object. The General Information Questionnaire, Nurse Transfer Tendency Scale, Work-Family Conflict Scale, and Job Burnout Inventory in Nurses were used for investigation. The structural equation was used to analyze the influencing factors of pediatric nurses ' transfer tendency. A total of 747 questionnaires were returned including 728 valid questionnaires with the valid response rate of 97.46%. Results:Among 728 pediatric nurses, the score of Nurse Transfer Propensity Scale was (11.64±2.24) , and the score of Work-Family Conflict Scale was (52.92±10.08) , and the score of Job Burnout Scale in Nurses was (62.48±19.14) . The structural equation model showed that work-family conflict and job burnout had a direct effect on the transfer tendency ( P<0.05) , and the number of children had direct and indirect effects on the tendency to transfer through work-family conflicts ( P<0.05) , and nurse-patient disputes and monthly night shift frequency had direct and indirect effects on the tendency to transfer through job burnout ( P<0.05) . Conclusions:Pediatric nurses have a high level of transfer tendency. The number of children, nurse-patient disputes, monthly night shift frequency, work-family conflicts, and job burnout have direct or indirect effects on the transfer tendency. Nursing managers should optimize staffing, improve the working environment, and reduce pediatric nurses ' transfer tendency.

Objective:To investigate the efficacy of continuous intravenous injection of hyaluronidase (HAase) combined with urokinase (UK) in the treatment of ischemia of hyaluronic acid (HA) arterial embolized skin flap.Methods:(1)Establish an animal model of superficial abdominal artery embolization with HA: 24 SD rats were used to make a square island flap (2 cm×2 cm, the width of the pedicle is 2 mm) on the left and right sides of the abdominal white line with superficial epigastric artery as feeding artery. The left and right flaps of rats were used as experimental group and control group. After the preparation of the experimental skin flap, HA 10 μl was injected into the superficial abdominal artery, while the self-control flap was only made and the vessels were peeled off without embolization.(2)The rats were randomly divided into A, B, C and D groups by drawing lots with 6 rats in each group, After the successful establishment of embolic animal model of 45 min, HAase(2 000 IU/kg)+ UK(50 000 IU/kg), HAase(2 000 IU/kg), UK(50 000 IU/kg) and normal saline were continuously infused through caudal vein with microinjection pump. The volume of solution in each group was 6 ml in 12 minutes. The general condition of rats and the skin color, edema, congestion and skin necrosis in the operation area of rats were observed, which were photographed and compared immediately, and at 3 days, 5 days and 7 days after embolization. The images were analyzed by Photoshop software, and the percentage of survival area of the flap after operation 7 days, was measured by pixel method. The percentage of survival area was compared by single factor analysis of variance (ANOVA) and head-to-head comparison by LSD- t test. Results:Immediately after operation, all the flaps in the four groups were pale and there was no obvious swelling. All the flaps showed different degrees of swelling within 3 days after operation, and the swelling basically disappeared within 3-5 days after operation, and gradually changed from dark red ecchymosis to dark purple or black. Seven days after operation, the necrotic area flap gradually hardened and its boundary was obvious. The skin flaps of the 4 groups showed different degrees of necrosis. The postoperative reaction of the flap was the slightest in group A, the performance of group B and C was similar, which were both between group A and group D, and there was the heaviest postoperative reaction in group D. At 7 days after operation, the percentage of survival area of flaps in groups A, B, C and D was 90.30%±5.95%, 52.63%±6.90%, 51.14%±5.95% and 7.70%±2.18%, respectively. The percentage of survival area of skin flap in group A was significantly higher than that in groups B, C and D (analysis of ANOVA: P<0.01; LSD- t test: P<0.01). Conclusions:Continuous intravenous infusion of HAase combined with UK can effectively alleviate the flap ischemia caused by HA artery embolism, increase tissue perfusion and increase the survival area of the flap.