1.Effects of IL-7 and IL-2 on CD4+CD25-T cells proliferation in vitro
Jie ZHANG ; Jingsheng HAO ; Junfu LI ; Longlong XING ; Zhiwen LIU ; Li ZHONG ; Wei LIU
Chinese Journal of Immunology 2014;(7):879-883,892
Objective:To explore the promoting effects of IL-7 and IL-2 on CD4+CD25-T cells proliferation in vitro and construct a stable culture system in vitro for CD 4+CD25+regulatory T cells from human umbilical cord blood.To compare the inhibiting effects between induced proliferated CD 4+CD25+Tregs and naturally isolated CD 4+CD25+Tregs on PBMCs functional activity.Methods:CD4+CD25-T cells and CD4+CD25+T cells were isolated from human umbilical cord blood mononuclear cells by magnetic activated cell sorting ( MACS) system and then expanded in vitro.Four different concentration levels of IL-7 combined with proper concentration of IL-2 were added as inducer and the efficiency and optimal concentration of IL-7 on inducing,CD4+CD25-T cells were analyzed via 4 different methods.Flow cytometry method was used to detect the changes of CD 4+CD25-T cells.The inhibitory effect of expanded CD 4+CD25+T cells on peripheral blood mononuclear cells (PBMCs) was tested by MTS.The expressions of Foxp3,IL-10 and TGF-βgenes in CD4+CD25+T cells were test by RT-PCR.Results:The CD4+CD25+T cells from each groups were expanded significantly after three weeks of culture.The results indicated that use of IL-7 combined with IL-2 resulted in the highest cell expansion comparing to the other groups.It was shown by the inhibitory test that the expanded CD 4+CD25+regulatory T cells could inhibit the proliferation of PBMCs ,but IL-7 induced CD4+CD25+regulatory T cells exerted weaker suppressor activity than natural regulatory T cells .Only IL-7 (4 ng/ml) and IL-2 (2 000 U/ml) induced CD4+CD25+regulatory T cells showed the strongest killing activity.Conclusion:We successfully expand CD4+CD25+regulatory T cells in vitro.The protocol is established in which the use of mAbCD 3/CD28 combined with IL-7 and IL-2 resulted in the highest cell expansion ,and intensely expressed cell phenotype of CD 4 and CD25.
2.Preoperative PSMA PET-CT guidance for patients with high-risk prostate cancer and its effect on postoperative positive margin
Penghe QUAN ; Changjiang YU ; Xiaozheng FAN ; Longlong ZHANG ; Jianhua JIAO ; Xing SU ; Shuaijun MA ; Peng WU ; Weijun QIN ; Xiaojian YANG
Chinese Journal of Urology 2021;42(9):706-711
Objective:To explore the preoperative 68Ga-PSMA PET/CT examination on the guidance of surgical strategies for high-risk prostate cancer patients and the influence of positive surgical margins after surgery. Methods:The clinical data of 118 patients with high-risk prostate cancer who underwent robot-assisted laparoscopic radical prostatectomy from June 2019 to December 2020 in Xijing Hospital of Air Force Military Medical University was retrospectively analyzed. 47 patients received 68Ga-PSMA PET/CT examination before surgery (study group), and 71 cases without 68Ga-PSMA PET/CT examination before operation ( control group). There was no statistically significant difference in the age [69 (63, 76) vs. 67 (64, 74) years], PSA [PSA≤20ng/ml: 9.91 (6.00, 13.67) vs. 11.64 (8.15, 15.44) ng/ ml, PSA> 20ng/ml: 66.53 (53.66, 195.30) vs. 63.18 (30.08, 148.05) ng/ml], preoperative clinical staging (T 2/≥T 3: 21/26 cases vs. 34/37 cases), and Gleason score [8 (7, 9) vs. 8 (7, 9) points] (all P>0.05) between study group and control group. Both groups underwent robot-assisted laparoscopic radical prostatectomy. The surgical plan was based on the PSMA PET/CT and MRI results in study group and control group respectively. First, ensure that all tumors are removed, and secondly, preserve the patient's urethral length as much as possible to ensure postoperative urinary control.If there is seminal vesicle invasion, expand the scope of resection as needed. If lymph node metastasis is shown, lymph node dissection is performed. For those with negative lymph nodes in imaging studies, if enlarged lymph nodes are found during the operation, lymph node dissection is also performed. After the operation, the perioperative results and surgical margins of the two groups were compared, and the correlation between the PSA value and the SUVmax value of prostate cancer tissue was analyzed. Results:The operations of the two groups were successfully completed, and there was no transfer to open surgery. The operation time of the study group was shorter than that of the control group [175 (155, 205) min vs. 205 (155, 235) min, P=0.003], and the positive rate of resection margin was significantly lower than that of the control group [23.40% (11/47) vs. 45.07%(32/71), P=0.017]. For patients with pathological stage ≥pT 3, the positive rate of surgical margins in the study group was significantly lower than that in the control group [30.77%(8/26) vs. 62.16%(23/37), P=0.014]. In the study group, 11 cases of PSMA-PET showed positive lymph nodes before operation, 10 cases were pathologically positive after operation (90.91%). PSMA-PET showed negative lymph nodes in 1 case, which was pathologically positive after operation. In the control group, 26 cases underwent lymph node dissection, and 16 cases (61.54%) were pathologically positive after operation. The preoperative PSA value of 47 cases in the study group was positively correlated with the SUVmax value of prostate cancer tissue ( r=0.579, P<0.01). Conclusions:Preoperative 68Ga-PSMA PET/CT for high-risk prostate cancer patients can guide the surgeon to optimize the surgical plan, reduce the positive rate of resection margins, and effectively remove the metastatic lymph nodes, which will benefit the patients.
3.Research advances in the mechanism of action of nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome in autoimmune hepatitis
Lifei WANG ; Longlong LUO ; Guojing XING ; Lixia LU ; Bin LI ; Jiucong ZHANG ; Xiaohui YU
Journal of Clinical Hepatology 2024;40(10):2092-2097
Autoimmune hepatitis (AIH) is chronic hepatitis caused by the attack of live cells by the immune system, and at present, the pathogenesis of AIH remains unclear. Inflammasomes are important components of innate immunity and are involved in a variety of pathophysiological processes. Studies have shown that inflammatory response associated with nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) plays an important role in the pathogenesis of AIH, which mainly mediates the release of proinflammatory factors and pyroptosis, thereby participating in the pathophysiological process of AIH. Therefore, the development and progression of AIH can be delayed by inhibiting the activation of NLRP3 inflammasomes, which provides new ideas for the prevention and treatment of AIH.
4.Role of mesenchymal stem cells and their exosomes in the treatment of drug-induced liver injury
Guojing XING ; Longlong LUO ; Lifei WANG ; Shunna WANG ; Xiaofeng ZHENG ; Lixia LU ; Jiucong ZHANG
Journal of Clinical Hepatology 2024;40(3):633-638
The incidence rate of drug-induced liver injury (DILI) is increasing year by year with unknown mechanisms, and the treatment methods for DILI mainly include drugs, liver support systems, and liver transplantation, all of which have certain limitations. Therefore, the search for safer and more effective treatment methods has become a research hotspot at present. Studies have shown that mesenchymal stem cells and their exosomes can alleviate liver injury by reducing liver inflammation, promoting hepatocyte proliferation and regeneration, inhibiting the apoptosis of hepatocytes, improving oxidative stress, and regulating immunity. This article briefly reviews the role of mesenchymal stem cells and their exosomes in the treatment of DILI, so as to provide a reference for further research.
5.Nanoplateletsomes restrain metastatic tumor formation through decoy and active targeting in a preclinical mouse model.
Longlong ZHANG ; Yuefei ZHU ; Xunbin WEI ; Xing CHEN ; Yang LI ; Ying ZHU ; Jiaxuan XIA ; Yiheng HUANG ; Yongzhuo HUANG ; Jianxin WANG ; Zhiqing PANG
Acta Pharmaceutica Sinica B 2022;12(8):3427-3447
Platelets buoy up cancer metastasis via arresting cancer cells, enhancing their adhesion, and facilitating their extravasation through the vasculature. When deprived of intracellular and granular contents, platelet decoys could prevent metastatic tumor formation. Inspired by these, we developed nanoplatesomes by fusing platelet membranes with lipid membranes (P-Lipo) to restrain metastatic tumor formation more efficiently. It was shown nanoplateletsomes bound with circulating tumor cells (CTC) efficiently, interfered with CTC arrest by vessel endothelial cells, CTC extravasation through endothelial layers, and epithelial-mesenchymal transition of tumor cells as nanodecoys. More importantly, in the mouse breast tumor metastasis model, nanoplateletsomes could decrease CTC survival in the blood and counteract metastatic tumor growth efficiently by inhibiting the inflammation and suppressing CTC escape. Therefore, nanoplatelesomes might usher in a new avenue to suppress lung metastasis.
6. The application of multi-point puncture D2-40, CD31, CD34 staining for autologous skin grafting in Kaposiform hemangioendothelioma
Longlong SUN ; Bin SUN ; Yaping XING ; Yuchun MA ; Hongzhao LEI ; Miaomiao LI ; Changxian DONG
Chinese Journal of Plastic Surgery 2019;35(11):1102-1106
Objective:
To explore the feasibility of application of multi-point puncture immunohistochemical to determine the invasion extent of the epidermis in Kaposiform hemangioendothelioma before surgery and choose the surgical method.
Methods:
From July 2013 to April 2016, 17 patients with Kaposiform hemangioendothelioma were admitted to the People′s Hospital of Zhengzhou University, including 7 males and 10 females. The first consultation age was 75.60±31.55 days. Preoperative multi-point pathological puncture D2-40, CD31, CD34 immunohistochemical staining was to test the invasion extent of the epidermis in Kaposiform hemangioendothelioma and to determine whether undergo autologous skin grafting. Skin graft survival areas were recorded postoperatively. Platelet values were examined after 1 week, 4 weeks, 12 weeks and 24 weeks.
Results:
In these 17 cases, 15 cases with epidermis and dermis unaffected was performed in situ grafting. 2 cases with the tumor invasion, superficial skin grafting were repaired. In these 15 cases of in situ grafting, platelet values returned to normal at 1 week after the operations, and platelet values were higher than 100×109/L at 4 weeks, 12 weeks and 24 weeks. Among them, 12 cases of epidermal survival area were greater than 90%, 2 cases were 70%-90%, and 1 cases was about 50%.
Conclusions
During the complete resection of Kaposiform hemangioendothelioma, the multi-puncture D2-40, CD31 and CD34 staining are used to determine invasion extent of the tumor to the skin, which can maximize the retention of normal tissue surface, reduce postoperative complications. This is a new multi-disciplinary surgical approach.
7.Mechanism of action of follicular helper T cells in autoimmune hepatitis
Longlong LUO ; Lifei WANG ; Guojing XING ; Ying ZHENG ; Lixia LU ; Chuyi LI ; Bin LI ; Jun MA ; Xiaohui YU ; Jiucong ZHANG
Journal of Clinical Hepatology 2024;40(7):1466-1469
Autoimmune hepatitis (AIH) is a type of chronic hepatitis caused by the attack of hepatocytes by the autoimmune system, and with the prolongation of disease course, it may gradually progress to liver cirrhosis and even hepatocellular carcinoma. Although great achievements have been made in the understanding and treatment of AIH, its etiology and pathogenesis still remain unclear. T cells play a crucial role in the development and progression of AIH, and by focusing on follicular helper T cells, this article elaborates on the research advances in follicular helper T cells in AIH, in order to provide new ideas and strategies for the clinical treatment of AIH.