This study intends to establish a colorectal cancer(CRC)organoid model,expand and isolate CRC-reactive tumor-infiltrating lymphocytes(TILs),screen tumor-specific T cell receptors(TCRs)and perform functional verification,in order to provide a technological platform and research foundation for the clinical transformation of individualized adoptive T-cell immunotherapy for colorectal cancer.An organoid model derived from colon cancer patient tissues was constructed using in vitro 3D culture techniques,which then subjected to HE staining and immunohistochemistry for detecting morphological characteristics and representative molecular expression.Subsequently,CRC organoids were co-cultured with TILs for sorting reactive TILs using flow cytometry,and the characteristics of reactive TCR clones was analyzed through single T cell receptor gene cloning technology.Furthermore,the function of TCRs was verified through cytotoxicity experiments.Morphological analysis and representative molecules(CK20 and CDX2)expression indicated that there is high similarity between colorectal cancer organoids and patient tumors.In the in vitro expanded and cultured TILs,colorectal cancer-reactive T cells with upregulated CD137 expression and increased IFN-γ secretion were screened out successfully,among which TCR2-T cells demonstrated superior tumor reactivity and in vitro tumor killing function.In conclusion,a platform for screening and function validation of reactive TCRs based on CRC-Org has been established,providing a technological platform for the translational application of individualized T-cell therapy for colorectal cancer.

The aim of this study is to explore the protective mechanism of Jiedu Tongluo Tiaogan decoction(JTTD)on islet cells of type 2 diabetic SD rats based on autophagy regulation.8-week-old SD male mice were fed with high-fat diet for 4 weeks and then induced with a single intraper-itoneal injection of streptozotocin 40 mg/kg.The Chinese medicine group was given the low(1.5 g·kg-1·d-1),medium(4.5 g·kg-1·d-1)and high(13.5 g·kg-1·d-1)doses of JTTD,and the western medicine group was given metformin hydrochloride(0.15 g·kg-1·d-1).Fasting blood glucose(FBG),glycated serum protein(GSP),triglycerides(TG),cholesterol(CHO),high-densi-ty lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),free fat(FFA)were measured.Moreover,the fasting serum insulin was determined by ELISA method and insulin resistance index(HOMA-IR)was calculated,glucose tolerance test was performed.The pancreatic tissues of SD mice were stained with HE,and the expression of Beclin-1 and LC3 were checked by immunohistochemistry,the expression of Beclin-1,LC3 and P62 was determined by Western blot,the expression of Beclin-1 and LC3RNA was determined by RT-PCR.The results showed that both the JTTD group and metformin hydrochloride group had better hypoglycemic and lipid-lowering effects(P＜0.05,P＜0.01).HE staining of pancreatic tissues showed that disorganized structure,unclear boundary,irregular arrangement,less cytoplasm,and lighter cytoplasmic staining or vacuo-lation in the islets in the JTTD group and metformin hydrochloride group were improved.In terms of autophagy regulation,immunohistochemistry,Western blot and RT-PCR,compared with the model group,the expression of Beclin-1 and LC3 mRNA was significantly increased(P＜0.01)and the level of p62 protein was decreased(P＜0.05,P＜0.01)in the JTTD group and metformin hydrochloride group.In conclusion,the JTTD can alleviate the trend of weight loss and improve ex-cessive drinking effectively in type 2 diabetic mellitus SD mice,as well as have better effects on im-proving glucose and lipid metabolism,moreover,it can alleviate the damage of pancreatic his-topathological structure and protect islet cells,which may be achieved by regulating the level of au-tophagy.

This study was performed to explore the impact of glutamine(Gln)on the anti-tumor immune response of CD8+ T cells and its mechanism.TCGA database was used to analysis the relationship between tumor Gln metabolism and the quantity and functionality of infiltrating CD8+ T cells.CRISPR/Cas9 was employed to knock down GLS expression in mouse MC38 cells,and a mouse tumor model was established.Flow cytometry was conducted to assess tumor proliferation,apoptosis,and the quantity and functionality of tumor-infiltrating immune cells.Lymphocytes isolated from health individuals were treated with Gln-deficient media,complete media or media supplemented with GSH,RSL3 in vitro.Then the apoptosis,the expression levels of GPX4,Lipid-ROS,and effector function protein of CD8+ T cells were detected by flow cytometry.Furthermore,RNA-seq was performed to analyze the differential gene expression on the Gln-depleted CD8+ T cells.Data showed that tumor Gln metabolism was inversely associated with the quantity and functionality of tumor-infiltrating CD8+ T cells.Low expression of GLS in MC38 cells could inhibit C57BL/6 tumor growth,decrease Ki-67 expression,promote casepase-3 expression,increase the amount of tumor-infiltrating immune cells,suppress PD-1,TIM-3,and LAG-3 expression,and enhance CD137,CD107a,IFN-γ and TNF-α expression in tumor-infiltrating CD8+ T cells.RNA-seq results indicated an upregulation of ferroptosis genes TFRC,HMOX1,CYBB and SLC7A11 in CD8+ T cells following glutamine deficiency.Gln deficiency led to lower CD137,CD107a,IFN-γ,GSH,GPX4 expression,increased Lipid-ROS level,and caused cell death in CD8+ T cells.Supplementation of GSH upregulated GPX4 expression,downregulated Lipid-ROS level,and increased IFN-γ secretion in CD8+ T cells.In conclusion,Gln deficiency inhibits the effector function of CD8+ T cells by inducing ferroptosis,and promotes tumor growth.

Objective:To investigate the effects of peripartum administration of low-dose corticosteroids or intravenous immunoglobulin (IVIG) on delivery outcomes in pregnant patients with primary immune thrombocytopenia (ITP).Methods:This prospective cohort study involved pregnant women (≥34 gestational weeks) who were diagnosed with ITP in Peking University People's Hospital from January 2017 to December 2021. Their platelet counts were between 20×10 9/L to 50×10 9/L without bleeding and none of them had been treated with any medications. All patients were divided into medication group (prednisone or IVIG) and platelet transfusion group based on their preference. Differences in vaginal delivery rate, postpartum hemorrhage rate and platelet transfusion volume between the two groups were compared using t-test, Wilcoxon rank sum test and Chi-square test. Binary logistic regression was used to investigate the factors influencing the rates of vaginal delivery and postpartum hemorrhage. Multiple linear regression was used to analyze the factors influencing the platelet transfusion volume. Results:A total of 96 patients with ITP were recruited with 70 in the medication group and 26 in the platelet transfusion group. The vaginal delivery rate in the medication group was higher than that in the platelet transfusion group [60.0% (42/70) vs 30.8% (8/26), χ 2=6.49, P=0.013]. After adjusted by the proportion of multiparae and the gestational age at delivery, binary logistic regression showed that the increased vaginal delivery rate in patients undergoing the peripartum treatment ( OR=4.937, 95% CI: 1.511-16.136, P=0.008). The incidence of postpartum hemorrhage in the two groups was 22.9% (16/70) and 26.9% (7/26), respectively, but no significant difference was shown ( χ 2=0.17, P=0.789). The median platelet transfusion volume was lower in the medication group than in the platelet transfusion group [1 U(0-4 U) vs 1 U(1-3 U), Z=－2.18, P=0.029]. After adjustment of related factors including the platelet count at enrollment, obstetrical complications and anemia, multiple linear regression showed that the platelet transfusion volume was also lower in the medication group (95% CI:0.053-0.911, P=0.028). Ninety-six newborns were delivered without intracranial hemorrhage. The overall incidence of neonatal thrombocytopenia was 26.0% (25/96). There was no significant difference in birth weight, and incidence of neonatal asphyxia or thrombocytopenia between the two groups. Conclusion:Peripartum therapy in ITP patients may increase vaginal delivery rate and reduce platelet transfusion volume without causing more postpartum hemorrhage.

OBJECTIVE: To explore the feasibility of using network system to carry out desktop health emergency response exercise for sudden poisoning incidents,and to understand the emergency response capability on sudden poisoning incident in Guangdong Province. METHODS: A chemical poisoning incident caused by drinking polluted water was simulated. Eight questions were designed in 4 sections including the emergency report,response,disposal and strain capacity to the emergency. A total of 10 teams from prefecture-level city in Guangdong Province answered through the desktop network exercise system designed by National Institute of Occupational Health and Poison Control. Meanwhile,the experts judged score using this system. RESULTS: Ten teams were familiar and adapted to the network system in a relatively short period of time. The scores of emergency report,response,disposal and strain capacity on emergency were(14. 9 ± 2. 3),(6. 0 ±2. 2),(20. 7 ± 4. 1) and(13. 2 ± 3. 5),respectively. The score rates of above 4 sections were(74. 5 ± 11. 5) %,(50. 4 ±18. 7) %,(43. 5 ± 8. 5) % and(66. 2 ± 17. 5) %,respectively. The score rate of emergency report was higher than that of response and disposal( P < 0. 05). The score rate of strain was higher than that of disposal( P < 0. 05). The total score of desktop exercises of the 10 teams was(55. 0 ± 8. 0),which was low on the whole. CONCLUSION: The network system can be used for desktop exercise for sudden poisoning incidents. There is a weak link in the health emergency response capacity of sudden poisoning incidents in Guangdong Province and the capability of emergency response and disposal should be strengthened.

Objective: To explore the effect of histone deacetylase (HDAC) inhibitor valproic acid (VPA) on the epithelial-mesenchymal transition (EMT) of colon cancer. Methods: With four colon carcinoma cell lines (DLD-1, HCT116, SW480 and HT29) as study subjects, the effect of different concentrations of VPA(0.5，5 mmol/L) on cell proliferation was detected by MTT assay. The expression level of EMT-related proteins (E-cadherin and vimentin) was detected by Western blotting; Phenotypic changes of E-cadherin and vimentin were detected by immunofluorescence staining; Cell migration and invasion ability was detected by wound healing and Transwell invasion assay, respectively. Results:After treated with different concentrations of VPAfor 48 h, low concentration of VPAmerely exerted any effect on the cell proliferation rate of four colon cancer cell lines, and thus was chosen as the experiment concentration; The results of Western blotting showed that the expression of E-cadherin was reduced (P<0.05) and vimentin was increased (P<0.05) in colon carcinoma cells by VPAtreatment (0.5 mmol/L); Immunofluorescence staining revealed membranous attenuation or nuclear translocation of E-cadherin and enhanced expression of vimentin after VPA treatment (0.5 mmol/L), and these responses occurred after 6 h and sustained until 24 h; Wound healing and Transwell invasion assay showed increased migration and invasion ability following VPA treatment (0.5 mmol/L). Conclusion: Low concentration VPA could induce the development of EMT in colon cancer cells by nuclear translocation of E-cadherin, and obviously enhance the migration and invasion ability of colon cancer cells; Thus, HDAC inhibitors, as a new type anti-cancer option, shall be carefully considered before their application in colon cancer.

Objective In order to acquaint with the prevalence of Tibetan sheep plague in this area, we conducted a serum epidemiological investigation of Tibetan sheep plague in Qinghai Province. Methods Indirect hemagglutination assay (IHA) and colloidal gold immunochromatography (GICA) were applied to test serum samples of Tibetan sheep and whole blood samples from jugular vein of Tibetan sheep were collected in 8 Prefectures of Qinghai Province from 2013 to 2016. Results A total of 86 positive Tibetan sheep serum samples with plague F1 antibody were detected by both methods, and the positive rate was 0.68％ (86/12710), the samples collected in Xinghai County Hainan Prefecture had the highest positive rate, which was 5.20％ (27/519). The Haixi Prefecture and Yushu Prefecture were historical epidemic areas, the positive rates were 0.65％(15/2313) and 0.26％(6/2293), respectively. Hainan Prefecture, Guoluo Prefacture and Huangnan Prefecture were newly confirmed epidemic areas, the positive rates were 1.61％ (28/1741), 1.01％ (15/1481), and 1.44％(19/1316), respectively. The antibody titers were 1:20 to 1:5120, the samples collected in Maqin County Guoluo Prefecture had the highest titer, namely 1 :5120. Conclusions In Qinghai Province, Tibetan sheep plague is endemic, and there are outbreaks in some regions. So we have to enhance the Tibetan sheep plague monitoring especially in Marmot plague epidemic area.

Objective: To investigate the effect of hypertensive disorder complicating pregnancy (HDCP) on the mortality and early complications of premature infants. Methods: The general clinical data of preterm infants with gestational age 24-36^{+ 6} weeks were collected from the cooperative units in the task group from January 1, 2013 to December 31, 2014.According to the severity of HDCP, the infants were divided into 4 groups: HDCP group, preeclampsia group, eclampsia group and non HDCP group, the mortality and major complications of preterm infants were compared, and the influencing factors were analyzed. Results: The mortality rate of preterm in the HDCP group was significantly higher than that of non HDCP group, and there was statistical significance (χ²=9.970, P=0.019). Eclampsia had a highest fatality rate (4.8%) in the early stage, compared with non HDCP group (2.2%), and the difference was statistically significant.Comparison of HDCP group (1.8%) and eclampsia group (3.2%) suggested that there was no statistically significant difference.The incidence of respiratory distress syndrome (RDS) in preterm in HDCP group was significantly higher than that of non HDCP group, and there was statistical significance (χ²=13.241, P=0.004). Eclampsia group showed the highest incidence (35.4%), compared with non HDCP group (16.2%), the difference was statistically significant, but compared with HDCP group (19.9%), preeclampsia group (17.1%), there was no significant diffe-rence.The incidence of bronchopulmonary dysplasia (BPD) in preterm in HDCP group was significantly higher than that of non HDCP group (χ²=9.592, P=0.022), the highest incidence showed up in eclampsia group (9.7%), compared with non HDCP group (2.0%) and HDCP group (1.7%), the difference was statistically significant.But there was no statistically significant difference, compared with preeclampsia group.As the degree of HDCP aggravated, the incidence of BPD gradually rose.There was no significant impact on necrotizing enterocolitis (NEC), retinopathy of prematurity (ROP), intraventricular hemorrhage (IVH) and sepsis of HDCP (χ²=7.054, 7.214, 0.358, 3.852; P=0.070, 0.065, 0.949, 0.278). Considering the overall outcome of the child, that was, whether the child died or survived, he had at least one complication, and HDCP had an effect on it (χ²=15.697, P=0.001), so the incidence increased while the degree of HDCP rose gradually.After adjusting gestational age, birth weight, sex, way of delivery, placental abruption and front placenta, prenatal hormonal, gestational diabetes, neonatal asphyxia and other factors, the results displayed that HDCP was the factor leading to the death of premature baby (OR=2.159, 95%CI: 1.093-4.266), and comparison between preeclampsia and eclampsia showed no statistical difference (P=0.714, 0.389); HDCP had no significant influence on RDS, BDP, ICH, NEC, ROP and sepsis. Conclusions HDCP leads to increased risk of premature death, but also leads to the increased incidence of RDS and BPD, but it had no obvious effect on NEC, ROP, IVH, sepsis and other complications.

Objective To construct a motivation goal framework for changes in transcultural self-management health behaviors among senile patients with chronic heart failure (CHF). Methods Totally 15 experts were selected to participate in Delphi consultancy from September 2016 to September 2017. Two rounds of questionnaire consultancy were conducted in terms of the proposed items for the motivation goal framework for changes in transcultural self-management health behaviors among senile CHF patients, with an unanimous understanding among the experts attained. Finally, the motivation goal framework for changes in transcultural self-management health behaviors among senile CHF patients was preliminarily constructed. Results The preliminarily constructed motivation goal framework for changes in transcultural self-management health behaviors among senile CHF patients included 3 primary items, 9 secondary items and 52 tertiary items. The authority coefficient, judgement coefficient and degree of familiarity of these experts were 0.930, 0.950 and 0.910, respectively. The return rate of valid questionnaires in the two rounds of expert consultancy reached 100%. Conclusions The motivation goal framework for changes in transcultural self-management health behaviors among senile CHF patients constructed under the transcultural nursing theory and the cross-theoretical model is scientific and practical, which provides reference for personalized, precise and normalized self-management education.

BACKGROUND: Because of the relatively similar size of organs to human and the physiological and structural similarities, the use of porcine as xenograft donors is progressing very actively. In this study, we analyzed the characteristics of porcine ear cartilage and evaluated its suitability as graft material in reconstructive and cosmetic surgery. METHODS: The auricular cartilage was harvested from two pigs, and subjected to histological examination by immunohistochemical staining. To determine the collagen content, samples were treated with collagenase and weight changes were measured. After sterilization by irradiation, the samples were grafted into rats and stained with Hematoxylin and Eosin and Masson Trichrome to observe inflammation and xenograft rejection. RESULTS: In IHC staining, extracellular matrices were mainly stained with type II collagen (20.69%), keratin sulfate (10.20%), chondroitin sulfate (2.62%), and hyaluronic acid (0.84%). After collagenase treatment, the weight decreased by 68.3%, indicating that about 70% of the porcine ear cartilage was composed of collagen. Upon xenograft of the sterilized cartilages in rats, inflammatory cells were observed for up to 2 months. However, they gradually decreased, and inflammation and reject-response were rarely observed at 5 months. CONCLUSION: The porcine ear cartilage was covered with perichondrium and cellular constituents were found to be composed of chondrocytes and chondroblasts. In addition, the extracellular matrices were mainly composed of collagen. Upon xenograft of irradiated cartilage into rats, there was no specific inflammatory reaction around the transplanted cartilage. These findings suggest that porcine ear cartilage could be a useful alternative implant material for human cosmetic surgery.
Animals ; Cartilage ; Chondrocytes ; Chondroitin Sulfates ; Collagen ; Collagen Type II ; Collagenases ; Ear Cartilage* ; Ear* ; Eosine Yellowish-(YS) ; Extracellular Matrix ; Hematoxylin ; Heterografts* ; Humans ; Hyaluronic Acid ; Inflammation ; Rats ; Sterilization ; Surgery, Plastic ; Swine ; Tissue Donors ; Transplants