ObjectiveTo observe the effects of high mobility group box-1 protein ( HMGB1 ) on function of murine spleen IL-10-producing dendritic cells (DCs) subset CD11clowCD45RBhighDCs.MethodsSplenic CD11clowCD45RBhighDCs and CD4 +T cells in Balb/c mice were purified by magnetic beads sorting.CD11clowCD45RBhighDCs were treated with various doses of HMGB1 (0,20,100,500ng/ml).Flow cytometry was used to determine expressions of CD11clowCD45RBhighDC surface molecules including CD40,CD80,CD86,Ⅰ-a/e and Toll-like receptor (TLR) 4.IL-10 level in CD11clow CD45RBhighDC culture supernatants was determined by ELISA method.The CD4+ T cells were divided into four groups: control group (without only treatment),untreated group (HMGB1-untreated CD11clow CD45RBhigh DCs plus CD4+T cells),high-dose HMGB1-treated group (500 ng/ml HMGB1-treated CD11clowCD45RBhigh DCs plus CD4 +T cells),high-dose HMGB1-treated + antibody 1 group (500 ng/ml HMGB1-treated CD11clowCD45RBhigh DCs plus IL-10 antibody and CD4+T cells),high-dose HMGB1-treated + antibody 2 group (500 ng/ml HMGB1-treated CD11clow CD45RBhigh DCs plus homotype IL-10 antibody and CD4 +T cells).IL-4 as well as interferon( IFN-γ) contents in CD4 + T cell culture supernatants were determined by flow cytometry.ResultsHMGB1 could markedly enhance the expressions of CD40,CD86 and TLR4 on CD11clowCD45RBhigh DC surface as well as level of IL-10 secreted by CD11clow CD45RBhigh DCs as compared with the non-HMGB1 treatment.Meantime,the secretion of IL-10 was HMGB1 concentration dependent.IFN-γ level in high-dose HMGB1-treated group was (279 ± 17) pg/ml,which was markedly lower than that in the untreated group [ (963 ± 11 ) pg/ml,P ＜ 0.05 \ ].IL-4 level in high-dose HMGB1-treated group was (372 ± 14) pg/ml,which was significantly higher than that in the untreated group [ (213 ± 10) pg/ml,P ＜0.05) \].ConclusionsHMGB1 promotes IL-10 production in CD11clowCD45RBhighDCs,induces the differentiation of CD4 +T cells to Th2 cells and downregulates the immune response via activating CD11clowCD45RBhigh DCs.

OBJECTIVE: To investigate the gene mutation occurved in AML patients with 29 kinds of fusion genes and 51 kinds of tumor gene. METHODS: Next-generation sequencing (NGS) was used to detected the 49 kinds of targeted gene. FLT3 internal tandem duplication (FLT3-ITD), CALR, NPM1 and CEBPA mutation were detected by DNA-based PCR and Sanger sequencing. Twenty-nine kinds of fusion genes were dected by multiplex nested RT-PCR. RESULTS: The total gene mutation rate was 91% (109/121) in all the 121 patients. On average, 2.1 mutated genes per patient were identified, among these 121 patients, coexistence of ≥ 3 mutations was frequent (34.7%). The most commonly mutated genes were NRAS (23.96%, n=29), followed by NPM1 (14.04%, n=17), CEBPA double mutations (14.04%, n=17), KRAS (11.57%, n=14)，FLT3-ITD (10.74%, n=13), CSF3R (10.74%, n=13), TET2 (9.92%, n=12) and IDH1 (9.1%, n=11). Overall, fusion genes were detected in 47 (37.3%) patients, including AML/ETO (n=12), CBFβ/MYH11 (n=11), PML/RARa (n=12), MLL rearranagement realated mutation MLL-X (n=10). TLS/ERG (n=1) and DEK/CAN (n=1) in an order of decreasing frequency. Patients with normal karyotype (NK)- AML exhibited more mutations in CEBPA, NPM1, TET2, RUNX1 and IDH1, comparing with abnormal karyotype patients. KRAS mutation in abnormal kayotype patients was significantly higher than that in normal kayotype patients (P=0.014). TP53 mutations were predominantly associated with complex cytogenetics (P=0.199). KRAS mutations were more frequent in core binding factor (CBF) acute myeloid leukemia (AML) and 11q23/MLL rearrangement leukemia, compared with NK-AML (P=0.006 and 0.003, respectively). KIT mutations predominated in CBF-AML (P=0.006). JAK2V617F mutations were detected in two patients and co-occurred with AML-ETO fusions. CONCLUSION At least one mutation is observed in more than 90% patients. On average, more than 2 mutated genes per patient are identified. Some gene mutations are associated with gene rearrangement.
Chromosomal Proteins, Non-Histone ; Genomics ; High-Throughput Nucleotide Sequencing ; Humans ; Leukemia, Myeloid, Acute ; Mutation ; Oncogene Proteins ; Poly-ADP-Ribose Binding Proteins ; Prognosis

Objective To explore the value of SPECT/CT in evaluation on long-term healing of limb fractures.Methods SPECT/CT imaging characteristics of 68 patients with limb fractures 6 months after treatment were analyzed retrospectively,and the self-healing property of fractures was evaluated.Compared with clinical follow-up results or surgical findings,the accuracy of SPECT/CT in evaluation on long-term healing of fractures was calculated.Results For all 68 patients with limb fractures,X-ray and CT were not able to determine whether the fractures would heal in the future.On SPECT/CT fusion images,radioactivity concentrations through the fracture lines and cross on both sides of fracture were found in 52 patients,which indicated that the fractures were in the process of healing and had the ability of healing in the future.Clinical follow-up confirmed that fractures then healed in 46 patients,while in 6 patients did not.Another 16 patients showed no nuclide concentrations on the fractures,indicated that the fractures were nonunion and had little ability of healing in the future,and were finally confirmed with surgery.The accuracy of SPECT/CT in evaluation on fracture healing was 91.18％ (62/68).Conclusion SPECT/CT imaging can accurately evaluate the long-term healing of limb fractures,which contributes to therapeutic planning in the clinic.

Objective To investigate the characteristics,treatment and prognosis of radiation-induced sympathetic chain injury.Methods We observed 6 cases with radiation-induced sympathetic chain injury and re-viewed the clinical features,treatment and prognosis of these patients. Results The median age of these 6 cases was 50 and the interval between radiotherapy to onset was 7.7 years.Dizziness was the first and most common symp-tom and dramatic fluctuation of blood pressure was the most common physical sign.Steroids might be considered for the treatment of radiation-induced sympathetic nerve injury. However,the therapeutic effect still needs to be proved.Conclusions Sympathetic nerve injury is an uncommon but dramatic symptom in NPC patients after radio-therapy.

Objective To study the effect of dopamine D1 receptor antagonist SCH23390 on the behavior of parkinsonian (PD) rats and on electrophysiological characteristics of the entopeduncular nucleus (EP).Methods Intracranial injection of 6-OHDA was conducted in 23 adult male Wistar rats to create PD animal models (experimental group) while the same amount of normal saline was injected in another 19 rats as a control group.(1) After intraperitoneal injection of SCH23390 at various concentrations of 0.010,0.015,0.020,0.025 and 0.030 mg/kg,the step frequency and the discontinuous moving frequency tests were carried out to determine the optimal concentration and time of intervention at 6 time points (10 main before,and 5～15,20～30,30～40,40～50 and 68～78 min after intervention) when all the rats were put on a treadmill at a speed of 8 r/min.(2) After recording electrodes were implanted into the EP in the rats,the signals of spikes in the EP in still and movement conditions were recorded simultaneously using the 16-Channel OmniPlex Neural Data Acquisition System.The spike signals collected were imported into Offiine Sorter to do cluster-sorting analysis and then into NeuroExplorer to analyze alterations in the firing pattern and firing rate of each type of neuron.(3) Frozen sections of the brain samples harvested by perfusion from the rats in the 2 groups were subjected to Nissl's staining and histological assay.Results (1) The behavioral tests showed that intraperitoneal injection of SCH23390 at 0.020 mg/kg at time points of 20～30 min and 30～40 min was optimal.The step frequency for the experimental group (24.47±1.35 step/min) was significantly decreased compared with that for the control group (30.77±2.06 step/min) (t=7.392,P=0.000).(2) After intervention by SCH23390,in movement and still conditions,the firing rate and variation coefficient of EP neurons were significantly higher in the experimental group than in the control group (P＜0.05).(3) The Nissl's staining showed a small number of lightly stained neurons were sparsely disposed in the injured cerebral substantia nigra and the histological assay found altogether 14 rats from which correct EP sites were collected.Conclusions This study demonstrated a negative regulating effect of dopamine D1 receptor antagonist SCH23390 on the behavior of PD rats,which may be exerted by changing the electrophysiological activity of EP.

Objective:To study the safety and efficacy of laparoscopic cholecystectomy (LC) in treatment of T2a and lower stages of gallbladder carcinoma.Methods:A retrospective study was performed on patients who were diagnosed with gallbladder cancer and underwent surgical treatment from January 2016 to January 2021 at Hunan Provincial People's Hospital. These patients were divided into the simple treatment group and the radical treatment group based on the surgical methods used. The simple treatment group consisted of 64 patients who underwent LC for accidental gallbladder cancers. The radical treatment group consisted of 30 patients who underwent laparoscopic radical cholecystectomy (LRC). The baseline characteristic of the two groups of patients were matched in a 1∶1 ratio using propensity score matching. After matching, there were 26 patients in each of the 2 groups. There were 7 males and 19 females in the simple group, with mean ± s. d. age of (60.6±9.6) years. There were 8 males and 18 females, with mean ± s. d. age (60.9±9.1) years in the radical treatment group. Blood loss, operation time, postoperative hospital stay, biliary leakage, acute pulmonary embolism, and incisional infection were compared between the two groups.Results:In the simple group, the operative time was (78.7±62.9) min, intraoperative blood loss was (10.7±11.6) ml and postoperative hospital stay was (4.4±2.6) d. These results were significantly better than those in the radical group with operative time (298.7±101.3) min, intraoperative blood loss (161.9±96.7) ml and postoperative hospital stay (9.9±4.0) d (all P<0.05). There were no significant difference in the postoperative complications and disease free survival rates between the two groups (both P>0.05). Conclusion:LC was safe and effective for treatment of T2a and lower stages of gallbladder cancer, and it could achieve a similar disease-free survival rate as LRC.

Objective:To observe the effect of noninvasive positive pressure ventilation (NIPPV) and high-flow nasal cannula oxygen therapy (HFNC) on the prognosis of patients with coronavirus disease 2019 (COVID-19) accompanied with acute respiratory distress syndrome (ARDS).Methods:A retrospective study was conducted in Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology when authors worked as medical team members for treating COVID-19. COVID-19 patients with pulse oxygen saturation/fraction of inspiration oxygen (SpO 2/FiO 2, S/F) ratio < 235, managed by medical teams [using S/F ratio instead of oxygenation index (PaO 2/FiO 2) to diagnose ARDS] from February to April 2020 were included. The patients were divided into NIPPV group and HFNC group according to their oxygen therapy modes. Clinical data of patients were collected, including general characteristics, respiratory rate (RR), fraction of FiO 2, SpO 2, heart rate (HR), mean arterial pressure (MAP), S/F ratio in the first 72 hours, lymphocyte count (LYM), percentage of lymphocyte (LYM%) and white blood cell count (WBC) at admission and discharge or death, the duration of dyspnea before NIPPV and HFNC, and the length from onset to admission. The differences of intubation rate, all-cause mortality, S/F ratio and RR were analyzed, and single factor analysis and generalized estimation equation (GEE) were used to analyze the risk factors affecting S/F ratio. Results:Among the 41 patients, the proportion of males was high (68.3%, 28 cases), the median age was 68 (58-74) years old, 28 cases had complications (68.3%), and 34 cases had multiple organ dysfunction syndrome (MODS, 82.9%). Compared with HFNC group, the proportion of complications in NIPPV group was higher [87.5% (21/24) vs. 41.2% (7/17), P < 0.05], and the value of LYM% was lower [5.3% (3.4%-7.8%) vs. 10.0% (3.9%-19.7%), P < 0.05], the need of blood purification was also significantly lower [0% (0/24) vs. 29.4% (5/17), P < 0.05]. The S/F ratio of NIPPV group gradually increased after 2 hours treatment and RR gradually decreased with over time, S/F ratio decreased and RR increased in HFNC group compared with baseline, but there was no significant difference in S/F ratio between the two groups at each time point. RR in NIPPV group was significantly higher than that in HFNC group after 2 hours treatment [time/min: 30 (27-33) vs. 24 (21-27), P < 0.05]. There was no significant difference in rate need intubation and hospital mortality between NIPPV group and HFNC group [66.7% (16/24) vs. 70.6% (12/17), 58.3% (14/24) vs. 52.9% (9/17), both P > 0.05]. Analysis of the factors affecting the S/Fratio in the course of oxygen therapy showed that the oxygen therapy mode and the course of illness at admission were the factors affecting the S/F ratio of patients [ β values were -15.827, 1.202, 95% confidence interval (95% CI) were -29.102 to -2.552 and 0.247-2.156, P values were 0.019 and 0.014, respectively]. Conclusion:Compared with HFNC, NIPPV doesn't significantly reduce the intubation rate and mortality of patients with COVID-19 accompanied with ARDS, but it significantly increases the S/F ratio of those patients.

Adult ; Cladribine/therapeutic use* ; Histiocytosis, Langerhans-Cell/drug therapy* ; Humans

Myocardial fibrosis （MF） is a common pathological manifestation of various heart diseases. Due to the non-renewable nature of myocardial cells， the occurrence of MF represents irreversible damage to the myocardium. Previous studies have suggested that fibroblast-mediated collagen deposition is the main mechanism of MF. Recent studies have found that there is an immune regulation mechanism in the heart itself， and macrophage activation/polarization plays an important role in MF. With the deepening of traditional Chinese medicine research， scholars have found that traditional Chinese medicine can interfere with MF by regulating the renin-angiotensin-aldosterone system （RAAS） system and the inflammatory process， repairing the extracellular matrix， managing oxidative stress， and maintaining the balance of autophagy. This process is closely related to the activation and M1/M2 polarization of macrophages. Throughout the MF process， macrophage activation is beneficial， but excessive activation will be harmful. In the early stage of MF， appropriate M1 macrophage polarization is conducive to activating immunity and removing harmful substances. In the middle and late stages of MF， appropriate M2 macrophage polarization is conducive to remodeling the damaged myocardium. If macrophage activation is excessive/insufficient， or the balance of M1/M2 macrophage polarization is broken， the effect changes from improvement to destruction. Traditional Chinese medicines that regulate the activation/polarization of macrophages have the effects of replenishing Qi and nourishing Yin， as well as regulating Qi and activating blood， but there are also some heat-clearing， dampness-drying， and detoxification products. Therefore， the occurrence of MF may be caused by Qi and Yin deficiency， damp heat accumulation， and Qi stagnation and blood stasis. By summarizing the biological processes involved in macrophage activation/polarization in MF， this paper expounded on the research progress of traditional Chinese medicine in regulating macrophage activation and M1/M2 polarization from different angles to improve MF， so as to provide a reference for the treatment of MF with traditional Chinese medicine.