Objective To investigate the effect of high expression of functional IDO trom donor DCs on transplantation rejection in mice heterotopic cardiac transplantation.Method Adenovirus vector containing IDO gene was used to infect donor (C57BL/6) DCs to obtain IDO+ DCs.Mouse heterotopic cardiac transplantation models were established (C57BL/6-BALB/c) and the following groups were set up:control group,DCs injection group and IDO+ DCs injection group.Survival time of the donor heart in all groups was observed.Meanwhile,donor hearts were harvested at 7 th day post-transplantation for different examinations,including pathological examination,mRNA expression of IDO by real-time PCR,and IDO protein expression by Western blotting.Peripheral blood of recipients was also harvested for T lymphocyte apoptosis rate examination by FACS.Result Cardiac allograft median survival time in control group,DCs injection group and IDO+ DCs injection group was 7,7.5,and 17.5 days respectively.IDO+ DCs treatment significantly prolonged the survival time of donor hearts (P＜0.01).Pathological grading was significantly decreased (P＜0.01).The CD3+ T lymphocyte apoptosis rate in peripheral blood of recipients in IDO+ DCs injection group was (46.50 + 5.02)％,significantly higher than the other two groups (P＜0.01).Both IDO mRNA and protein expression showed significant increase (P＜0.01).Conclusion The preoperative medication with IDO+ DCs injection to the recipients could induce T lymphocyte apoptosis and significantly suppress the rejection in mice heterotopic cardiac homotransplantation.

In present study,comparisons were carried out to develop and improve in vitro mauration(IVM) systems of goat oocytes. Oocytes were cultured in TCM-199 supplemented with(1)10% sera (either estrous goat serum (EGS) or fetal bovine serum(FBS)) + 20 mg/L luteinizing hormone (LH) + 10 mg/L follicle stimulating hormone (FSH) + 1 mg/L estradiol (E2); (2)10% EGS with different gonadotropins(LH: FSH) at a concentration of 5 mg/L: 0.5 mg/L or at 20 mg/L: 10 mg/L with0. 075 IU/mL human menopausal gonadotropin(HMG),1 mg/L estradiol 17β; (3)10% EGS+ 0. 075 mg/L HMG+10-20 μg/EGF. The culture was also performed by M199 supplemented with 10% EGS+0. 075 mg/L HMG+ 10-20 μg/L EGF into ultra-filtrated water which was derived either from self-producing or from purchased for use. The oocytes were cultured at 38 ℃,5% CO2 in air for 24 h,and the meterphase Ⅱ stage oocytes were examined under dissecting microscope. The results showed that EGS was better than FBS in supporting goat oocyte IVM. An addition of HMG in M199 could improve oocyte maturation and induce a higher percentage of metaphase Ⅱ oocytes compared to gonadotropins. 10-20 μg/L EGF improved goat oocyte maturation but the influence was not significant. Fresh,high quality water was vital for oocyte IVM. In conclusion,under our conditions with IVM ,the best result in maturation of goat oocyte has been M199 supplemented with 10% EGS+0.075 IU/mL HMG+10-20 μg/L EGF and prepared in fresh purified water.

Type B aortic dissection is a life-threatening aortic disease.With the development of clinical classification and diagnostic method,the mortality of type B aortic dissection has been greatly decreased.In respect of treatment,endovascular repair due to minimally invasive advantages become the first choice for the treatment of complicated type B aortic dissection.For non-complicated type B aortic dissection,endovascular repair also gradually replace drug treatment,and showed good efficacy.Open surgery is only available for patients who ate not suitable for endovascular repair or repair failure or patients with connective tissue disease.

Objective:To investigate the effects of histone deacetylase 3 (HDAC3) on the pyroptosis of breast cancer (BC) cells via regulating miR-625/anti-silencing function 1B (ASF1B) and its mechanism.Methods:The expression level of HDAC3, miR-625 and ASF1B in BC tissue, adjacent normal tissue, BC cell lines (T47D, MCF7 and MDA-MB-231) and human normal breast epithelial cell MCF-10A was detected by qRT-PCR. The expression level of cell pyroptosis related protein NLRP3, Caspase-1 and GSDMD was detected by Western blot. The expression level of IL-18 and IL-1βwere detected by ELISA. ChIP experiment was used to determine the interaction between HDAC3 and miR-625. The dual luciferase reporter assay was used to verifiy the targeted regulation between miR-625 and ASF1B.Results:Compared with adjacent normal tissue and MCF-10A cells, the expression of HDAC3 and ASF1B was increased and the expression of miR-625 was decreased in BC tissue and cells (all P<0.05) . Compared with si-NC group, the protein expression level of NLRP3, Caspase-1 and GSDMD in si-HDAC3 group was increased, and the concentration of IL-18 and IL-1β in cell culture supernatant was increased (all P<0.05) . HDAC3 inhibited the expression of miR-625 by binding to the promoter region of miR-625 ( P<0.05) . Compared with si-HDAC3+miR-NC group, The expression of NLRP3, Caspase-1, GSDMD, IL-18 and IL-1β in si-HDAC3+miR-625 inhibitor group was decreased (all P<0.05) . ASF1B was confirmed as a target gene of miR-625, the level of pyroptosis related factors in si-HDAC3+pcDNA3.1-ASF1B group was significantly lower than that in si-HDAC3 + pcDNA3.1-NC group. Conclusion:HDAC3 up regulates the expression of ASF1B by inhibiting miR-625, and then inhibits BC cell pyroptosis.

Objective To investigate the effects of probiotics and synbiotics on inflammation and microbiota of acute colitis in mice.Methods C57BL/6J mice were divided into 4 groups randomly.Each group had 10 mice and was given 2.5％ dextran sulfate sodium (DSS) drinking water for 5 days other than the blank control group.Except for model control group,other two groups were administrated with probiotics and synbiotics,respectively.Probiotics was composed of Lactobacillus acidophilus,Lactobacillus rhamnosus and Bifidobacterium lactis,while synbiotics was composed of the aforementioned probiotics,inulin and galactooligosaccharide.Feces of different periods and mucosa samples were collected to analyze the differences of enteric flora by 16s rDNA sequencing.Results (1) Pathological scores in probiotics group and synbiotics group were 5.40±2.79 and 7.25±2.87,respectively,which were significantly lower than those in the model control group with scores 27.00 ± 7.94.Model control group,probiotics group and synbiotics group showed lower flora diversity,increased Bacteroides and decreased Faecalibacterium than blank control group.The mucosal microbiota was different from fecal flora in abundance and species for each group,and Mucispirillum was more common in mucosa.Conclusions Probiotics and synbiotics alleviate the inflammation of acute colitis in mice.Imbalance of beneficial genera to harmful genera is the characteristic of acute colitis.Supplementation of probiotics and synbiotics contributes to regulating the balance of intestinal microbiota.

Objective To improve the knowledge and early diagnostic rate of primary small intestinal tumor.Methods From August 2012 to August 2017,hospitalized patients with pathological diagnosis of primary small intestinal tumor (excluding duodenal neoplasm) from Peking Union Medical College Hospital were retrospectively enrolled.The data of clinical manifestations,laboratory examinations,imaging,endoscopy examination,pathological findings and treatment were collected and analyzed.Results A total of 180 patients with primary small intestinal tumor were enrolled.The common clinical manifestations included abdominal pain (76 cases,42.2 ％),gastrointestinal bleeding (64 cases,35.6％),and abdominal distension (30 cases,16.7％),and 22 (12.2％) patients had no overt clinical symptoms.The sensitivity of carbohydrate antigen 19-9 (CA19-9) in the diagnosis of small bowel adenocarcinoma was 57.1％ (12/21).The diagnostic rates of computed tomography enterodysis (CTE),positron-emission computed tomography (PET)/computed tomography (CT),and abdominopelvic enhanced CT were 96.5％ (83/86),100.0％ (29/29),and 91.5％ (43/47),respectively.The diagnostic small intestinal tumor patients of barium radiography (14 cases),abdominopelvic magnetic resonance imaging (MRI) (eight cases),small bowel endoscopy (18 cases) and capsule endoscopy (eight cases) were seven,six,fifteen and six cases,respectively.Among 180 patients,14 (7.8％) patients were considered gynecological tumors by imaging examination before surgery,seven (3.9％) patients underwent emergency operation because of intestinal obstruction,four (2.2％) patients underwent emergency surgery due to gastrointestinal bleeding,and four (2.2％) patients underwent emergency surgery because of intestinal perforation.Histopathological type included gastrointestinal stromal tumor (117 cases,65.0％),lymphoma (25 cases,13.9％) and adenocarcinomas (21 cases,11.7％).Except seven patients with intestinal lymphoma who received chemotherapy,the rest 173 patients underwent surgical resection.Conclusions Primary small bowel tumor has no specific clinical manifestations.It should be alert on patients without positive findings by regular gastroendoscopy and colonendoscopy examination but with symptoms of abdominal pain,gastrointestinal bleeding and intestinal obstruction.CTE should be the first choice for patients with symptoms but unclear diagnosis.

Objective To investigate the effectiveness and safety of volumetric modulated arc therapy (VMAT) hypofractionated radiotherapy and intensity modulated conformal radiotherapy technique (IMRT) conventional fractionated radiotherapy after breast cancer radical operation.Methods Eighty-five patients with breast cancer modified radical operation admitted and treated in this hospital from March 1,2021 to De-cember 30,2021 were selected as the research subjects and divided into the VMAT group (n=41) and the IM-RT group (n=42) according to the random number table method.The VMAT group adopted the hypofrac-tionated radiotherapy,with the single fractionated dose of 2.9 Gy/frequency and radiotherapeutic total dose of 43.5 Gy/15 frequencies;the IMRT group adopted the IMRT conventional fractionated radiotherapy,with the single fractionated dose of 2.0 Gy/frequency and radiotherapeutic total dose of 50.0 Gy/25 frequencies.The planning target region V95,V110,conformity index,homogeneity index,treatment time,V5,V20,V30,average dose (Dmean) in the affected side lung,humeral head Dmean and heart V30,Dmean were compared between the two groups.Meanwhile,local recurrence,distant metastasis,disease-free survival and acute and chronic radiation injury were compared between the two groups.Results Compared with the IMRT group,V95 in the VMAT was higher,V110 and homogeneity index were lower,the treatment time was shorter,V5 in the affected lung,Dmean and Dmean in the affected humeral head were lower,V30 in the affected lung was higher,heart V30 in the left side breast cancer was lower,heart Dmean in the right side breast cancer was lower,and the differences were statistically significant (P<0.05).All patients survived without local relapse.The distant metastasis rate and disease free survival rate had no statistical difference between the two groups (P>0.05).Follow up lasted for 12 months,the incidence rates of grade Ⅰ-Ⅱ acute radiodermatitis,radiation esophagitis,chronic radioder-matitis and radiation pneumonia had no statistical differences between the two groups (P>0.05).The inci-dence rate of grade Ⅰ-Ⅱ shoulder dysfunction in the VMAT group was lower than that in the IMRT group with statistical difference (P<0.05).No grade Ⅱ and above acute and chronic radiation injury in the two groups occurred.Conclusion VMAT hypofractionated radiotherapy after breast cancer radical operation is safe and effective.

Objective:To compare the efficacy and safety of azacitidine combined with venetoclax or CAG regimen in the treatment of newly treated elderly patients with acute myeloid leukemia (AML).Methods:A retrospective cohort study was conducted. The clinical data of 34 newly treated elderly patients with AML treated in the Fifth People's Hospital of Datong from May 2018 to August 2023 were retrospectively analyzed. According to the treatment regimen, all patients were divided into venetoclax group (azacitidine + venetoclax, 17 cases) and CAG group (azacitidine + CAG regimen, 17 cases). The clinicopathological characteristics, efficacy, adverse reactions and survival of the both groups were compared.Results:There were no statistically significant differences in the clinical data of both groups (all P > 0.05). The complete remission (CR) rate and the objective response rate (ORR) in venetoclax group were higher than those in CAG group [CR: 70.6%(12/17) vs. 47.1% (8/17); ORR: 82.4% (14/17) vs. 64.7% (11/17)],while the differences in CR and ORR were not statistically significant (χ 2 = 2.00, P = 0.163; χ 2 = 2.00, P = 0.244). The follow-up time[ M ( Q1, Q3)] was 25.4 months (7.2 months, 60.3 months). At the end of follow-up, 19 of 34 patients survived (13 cases in venetoclax group and 6 cases in CAG group); 15 died (4 cases in venetoclax group and 11 cases in CAG group). The median overall survival (OS) time was 14.22 months (95% CI: 8.2-60.3 months) and 10.56 months (95% CI: 7.2-50.2 months), respectively in venetoclax group and CAG group;the median progression-free survival (PFS) time was 9.97 months (95% CI: 5.4-40.5 months) and 6.82 months (95% CI: 5.0-36.2 months), respectively, and there were no statistically significant differences in OS and PFS between the two groups (all P > 0.05). Grade 3-4 hematological adverse reactions occurred in 16 and 14 patients in venetoclax group and CAG group, respectively. There were no significant differences in granulocyte deficiency time, platelet deficiency time, infection and bleeding incidence between the two groups (all P > 0.05). Conclusions:Azacitidine combined with venetoclax or CAG regimen have better clinical efficacy and safety for newly treated elderly patients with AML.

OBJECTIVETo study the suppressive effect of indoleamine 2, 3-dioxygenase on transplantation rejection in mice heterotopic cardiac transplantation.

METHODSAdenovirus vector containing IDO gene was used to infect donor (C57BL/6) DC to obtain IDO(+)DC. Mouse heterotopic cardiac transplantation models were established (C57BL/6-BALB/c) and the following groups were set up, including the control group, DC injection group, TC injection group, IDO(+)DC injection group and co-injection group of IDO(+)DC and TC, 12 donors and 12 recipients in each group.Survival time of the donor heart in every group was observed. Meanwhile, donor hearts were harvested 7 days post transplantation for different examinations, including pathological examination, mRNA expression of IDO through real-time PCR, IDO protein expression through Western blot. Peripheral blood of recipients was also harvested for CD3(+)T lymphocyte apoptosis rate examination through fluorescence-activated cell sorting.One-way ANOVA and Kaplan-Meier Survival Analysis were used for statistic analysis of IDO expression, CD3(+)T lymphocyte apoptosis rate and survival time of the donor heart respectively.

RESULTSCadiac allograft median survival time of each group were 7.0, 7.5, 11.0, 17.5, 24.0 days respectively. Compared with control and DC injection group, IDO(+)DC, TC and co-injection group significantly prolonged the survival time of donor hearts (t = 3.523-8.449, P < 0.01). Both IDO mRNA and protein expression showed significant increase(t = 5.974-16.176, P < 0.01). The CD3(+)T lymphocyte apoptosis rate was also significantly increased (t = 6.324-38.120, P < 0.01). Compared with IDO(+)DC or TC group alone, co-injection group significantly prolonged the survival time of the donor heart (t = 5.971 and 2.831, P < 0.05). Both IDO mRNA and protein expression showed significant increase (t = 2.853-15.194, P < 0.01).Furthermore, the CD3(+)T lymphocyte apoptosis rate was significantly increased as well (t = 26.069 and 7.643, P < 0.05).

CONCLUSIONSSuppressive effect of co-injection of IDO(+)DC and TC is much more effective than administration of IDO(+)DC or TC alone, which suggests that IDO achieved immune suppressive effect through the pathway of tryptophan depletion and accumulation of TC.

Animals ; Gene Transfer Techniques ; Graft Rejection ; drug therapy ; Heart Transplantation ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL

Objective:To establish a model C-GALAD for detecting hepatocellular carcinoma (HCC) from the chronic liver disease and healthy people based on the serum markers.Methods:A clinical cohort including 229 hepatocellular carcinoma patients, 2 317 patients with chronic liver disease and 982 healthy people, was retrospectively collected from eight hospitals or physical examination institutions from April 2018 to October 2020. The data were divided into a training set and a testing set by stratified sampling with a 6∶4 ratio. A predictive model was established on the training set using a logistic backward regression method and validated on the testing set. In addition, clinical data from March to July 2021 in Beijing You′ an Hospital affiliated to Capital Medical University, including 84 patients with liver cancer and 204 patients with chronic liver disease collected were used for external independent validation of the model. The receiver operating characteristic curve (ROC) area under curve (AUC), the sensitivity and the specificity were used to evaluate the effectiveness of the model.Results:Through the logistic backward regression method, the seven signatures including age, gender, alpha-fetoprotein (AFP), alpha-fetoprotein alloplasm-3 ratio (AFP-L3%), des-gamma-carboxyprothrombin(DCP), platelet (PLT) and total bilirubin (TBIL) were selected as risk factors in the detection model. The area under the ROC curve (AUC) of the model on the testing set was 0.954, with an 88.04% sensitivity and a 94.85% specificity, and the AUC of model on the external independent validation set was 0.943, with an 89.29% sensitivity and a 90.2% specificity, which were better than other published models.Conclusion:The C-GALAD Ⅱ model can accurately predict the risk of hepatocellular carcinoma occurrence, and thus provide a trustworthy diagnosis method of hepatocellular carcinoma.