An introduction to the classified management of such trials and clinical application,documents submission for ethical review,operating procedures and review key points,as well as the problems found in the review.These efforts aim to provide guidance to the review of medical technique clinical trials,and to promote ethical supervision for new medical techniques.

Objective To knockout Rag2 and IL2rg genes and construct severe combined immunodeficiency mice based on CRISPR/Cas9 technology. Method Design and synthesis of 25 bp sgRNA were made according to the Rag2 and IL2rg sequences in Genbank. After annealing, sgRNA was cloned into pX330 vector. Recombination plasmid Rag2?sgRNA, IL2rg?sgRN and Cas9 were then transcribed into RNA, these RNA were microinjected into zygotes and the zygotes were transplanted into recipient ICR mice. F0 founders were born and mutated F0 founders mated with wild type mice to obtain F1 generation heterozygous mice. Mutated F1 mice were crossed and got F2 generation homozygous mice. Genotype and phenotype of the knockout mice were identified by sequencing, flow cytometry and xenograft model. Results Rag2?sgRNA and IL2rg?sgRNA recombination plasmids were constructed and transcribed into RNA. After microinjection and mat? ing, F0 founders were born and F2 homozygous mice were obtained. The results of sequencing showed that there were two types of genotype in IL2rg gene, 10 bp or 11 bp deletion;however, there was only one genotype in Rag2 gene, which was 8 bp deletion. Compared with wild?type BALB/c mice, the number of CD3 +, B220 + and NKp46 + cells in peripheral blood of the knockout mice was reduced significantly. After inoculation of human breast cancer cell line SKBR?2HL cells, tumor size in the xenograft mouse model was increased gradually along with time extension. Conclusion CRISPR/Cas9 is an efficient way to mutate Rag2 and IL2rg gene in mice in vivo, leading to aberrant T cells, B cells and NK cells.

Objective To construct miRNA-29b1 gene knockout mice based on CRISPR/Cas9 technology. Methods To design and synthesize sgRNA according to the miRNA-29b1 sequence in Genbank .sgRNA and Cas9 were transcribed to RNA in vitro, these RNA were then microinjected into zygotes of C 57BL/6 mice.After mouse birth, the genome DNA was extracted and sequenced to identify its genotype; meanwhile , real-time PCR was used to assay the expression of miRNA-29b1 in the heart, liver, spleen, lung and kidney of mutated mice .Result A 20 bp sgRNA targeted on miRNA-29b1 was synthesized and transcribed to RNA with Cas 9.After microinjection, miRNA-29b1 gene-mutated mice were obtained.The sequencing results showed that there were two types of genotype for the mutated mice , one was 10 bp deletion, and another was 23 bp deletion accompanied with a 3 bp insertion.Compared with the wild-type mice, the expression of miRNA-29b1 in the heart, liver, spleen, lung and kidney was reduced significantly .Conclusions miRNA-29b1 gene knockout mice are constructed successfully by using CRISPR /Cas9 technology.

Objective:To study the performance of immune reconstitution in patients with chimeric antigen receptor (CAR)-T cell immunotherapy bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:A total of 61 patients with acute B lymphocytic leukemia (B-ALL) who received CAR-T cell bridging allo-HSCT in Beijing Lu Daopei Hospital from August 2018 to December 2021 were enrolled, and the clinical medical records of the above patients were retrospectively analyzed. The average age was 14 (7, 30) years old, including 39 males and 22 females. 32 patients were treated with CAR-T cell immunotherapy(CAR-T Group) and 29 didn't with CAR-T cell immunotherapy(non-CAR-T group). The follow-up period was 561 (235,784) days. Multicolor flow cytometry was used to detect the peripheral blood lymphocyte subsets, i.e. total lymphocytes, T lymphocytes, helper T cells, cytotoxic T cells, B lymphocytes, NK cells, and Treg cell counts before transplantation and 1, 2, 3, 6, 8, 10, and 12 months after transplantation, to evaluate the immune reconstitution performance post allo-HSCT.Results:Serum globulin before transplantation: The IgA level in the CAR-T group was 0.18 (0.06, 0.49) g/L, which was lower than that of 1.03 (0.63, 1.56) g/L in the non-CAR-T group ( U=103.5, P<0.001). The IgG level in the CAR-T group was 5.54 (4.04, 7.09) g/L, lower than that of 6.78 (5.27, 9.26) g/L in the non-CAR-T group, ( U=1 298.5, P=0.017), and the IgM level in the CAR-T group was 0.18 (0.05, 0.30) g/L, lower than that of 0.40 (0.26, 0.71) g/L in the non-CAR-T group ( U=166.0, P<0.001). In the CAR-T group before transplantation, the absolute count of total lymphocyte in peripheral blood was 833.00 (335.00, 1 727.50) /μl, lower than that of 1 052.00 (545.75, 1 812.50) /μl in the non-CAR-T group ( U=404.0, P<0.001). The absolute count of T lymphocyte in the CAR-T group before transplantation was 686.00 (233.00, 1 307.00)/μl, lower than that of 860.00 (391.00, 1 419.75) /μl in the non-CAR-T group ( U=406.0, P<0.001). The absolute count of helper T lymphocytes in the CAR-T group was 146.00 (40.50, 327.50) /μl, lower than that of 162.50 (66.00, 384.75) /μl in the non-CAR-T group ( U=494.0, P=0.002). The absolute count of cytotoxic T lymphocytes in the CAR-T group was 343.00 (56.50, 924.00) /μl, lower than that of 478.00 (143.50, 992.25) /μl in the non-CAR-T group ( U=483.5, P=0.001). The absolute count of B lymphocytes in CAR-T group was 22.00 (6.00, 186.00) /μl, lower than that of 33.00 (8.00, 220.00) /μl in the non-CAR-T group ( U=498.0, P=0.002). And when two groups of patients were monitored after transplantation, there was no statistical difference in absolute cell counts of each immune cell subpopulation( P>0.05). Comparing the clinical features of the two groups, the pre-transplant history of the CAR-T group was 981.00 (368.50, 1 514.75) d, longer than that of 323.00 (167.50, 450.50) d in the non-CAR-T group ( U=263.0, P=0.004). The dose of rabbit anti-human thymic immunoglobulin (ATG) in the pretreatment protocol of patients in the CAR-T group was 5.00 (5.00, 7.50) mg/Kg, lower than that of 7.00 (5.00, 7.50) mg/kg in the non-CAR-T group ( U=288.5, P=0.018). The infusion dose of CD34 +cells in the CAR-T group was 5.91 (4.23, 6.02) ×10 6/kg, higher than that of 4.51 (4.00, 5.93)×10 6/kg in the non-CAR-T group ( U=291.0, P=0.012). The duration of the application of cyclosporine after transplantation in the CAR-T group was 167.00 (119.25, 299.50) d, which was shorter than that of 197.00 (102.50, 450.50) d in the non-CAR-T group ( U=421.0, P=0.001). Conclusions:For patients in CAR-T group with low immune function before transplantation, it may be possible to make them comparable to non-CAR-T group in immune reconstitution state by reducing the dose of pretreatment ATG, increasing the counts of CD34 + cells infusion in the graft, and discontinuing cyclosporine as soon as possible after transplantation.

Objective:To investigate the current situation of motivations for hospice care volunteerism among undergraduate nursing students, and to analyze its influencing factors.Methods:Using the convenient sampling method, a total of 575 nursing students from Nursing Department of Xinjiang Medical University were selected as the research objects in August 2022. They were investigated using general information questionnaire, Chinese version of Inventory of Motivations for Hospice Palliative Care Volunteerism Scale, The Palliative Care Quiz for Nursing, the Organizational Climate Scale and Prosocial Tendencies Measure Scale.Results:The score of Motivations for Hospice Palliative Care Volunteerism Scale for 575 nursing students was (87.32±20.54) , score of Palliative Care Quiz for Nursing was (7.89±3.44) , score of Organizational Climate Scale was (49.35±10.33) and the score of Prosocial Tendencies Measure scale was (96.71±18.25) . Qualities and abilities that college student volunteers should possess, access to hospice care, pro-social tendencies, and knowledge of palliative care were influential factors in nursing students' motivation to volunteerism for hospice care ( P<0.05) , which could explain 19.9% of the total variation. Conclusions:The motivations for hospice care volunteerism of undergraduate nursing students is in a medium and above level. Nursing educators should strengthen education and training related to nursing students' participation in hospice care, so that nursing students will participate in voluntary service activities through their own professional advantages, strengthen the voluntary team of hospice care, and promote the development of voluntary hospice care services.

Objective:To explore the role and mechanism of nuclear receptor subfamily 4 group A member 1 ( NR4A1) in suppressing cisplatin nephrotoxicity. Methods:The expression of NR4A1 gene in renal cell subpopulations was analyzed using the "Tabula-muris" single cell transcriptome sequencing database. NR4A1 gene was over-expressed by lentivirus infection in HK-2 cell line and primary renal proximal tubular epithelial cells. Cell counting kit-8 was used to detect the cytotoxicity of cisplatin. The cell death ratio was analyzed using propidium iodide (PI) staining by flow cytometry. The expression of NR4A1 and nuclear factor erythroid 2-related factor 2 ( NRF2) was detected by real-time fluorescent quantitative PCR and Western blotting. Ferroptosis was analyzed by detecting the contents of malondialdehyde (MDA), oxidized glutathione (GSSG) and lipid reactive oxygen species (ROS). Results:The single cell transcriptome sequencing database showed that NR4A1 gene was the lowest expression in renal proximal tubular epithelial cell subsets. Cisplatin (50 μmol/L or 100 μmol/L) could significantly induce MDA, GSSG and lipid ROS production in renal proximal tubular epithelial cells (all P<0.01), and higher cisplatin concentration accompanied with a more increase of MDA, GSSG and lipid ROS. Compared with the control HK-2 cells, the lipid ROS content and iron ion content of HK-2 cells over-expressing NR4A1 were significantly lower (all P<0.01), and the over-expression of NR4A1 inhibited cisplatin-induced cytotoxicity and ferroptosis in renal proximal tubular epithelial cells. Mechanistically, NR4A1 up-regulated the expression of anti-ferroptosis gene NRF2 in proximal renal tubular epithelial cells ( P<0.01). Furthermore, single cell data analysis showed that, similar to the expression of NR4A1 in renal tissue subsets, NRF2 was also the lowest in renal proximal tubular epithelial cells. Conclusions:Cisplatin can induce ferroptosis of renal proximal tubular epithelial cells in a dose-dependent manner. NR4A1 can inhibit cisplatin-induced ferroptosis by up-regulating NRF2 in renal proximal tubular epithelial cells, thereby alleviating the cytotoxicity of cisplatin.

Objective:The purpose of this study is to establish the trajectory of targeted grafting for facial fat compartment based on anatomical research, and then bring it to clinical practice.Methods:The boundary of fat compartment and the relationship of adjacent vessel and nerve were clarified through autopsy. The recommended injection points and trajectory for targeted fat grafting were established on the anatomical findings. Retrospective clinical data of facial rejuvenation of 46 patients through targeted fat grafting were collected from June 2017 to June 2019 in Shanghai Ninth People’s Hospital. The result of 3D scanning were analyzed to evaluate the survival rate of fat grafts.Results:There were subcutaneous superficial fat compartments in the frontal region, and there were both deep and superficial fat compartments in the temporal and middle face. According to the anatomical characteristics, a targeted fat grafting technique was established with the frontal hairline and the oral commissure corner mucosa as the entry points. In the clinical study, 46 patients were evaluated by 3D scanning 6 months after the last fat grafting. The amount of fat grafts in the temporal region was (17.84±8.47) ml and (11.2±2.44) ml was left after operation, and the survival rate was 63%. The amount of fat grafts in mid-face was (26.81±10.36) ml and (16.09±4.48) ml was left after operation, and the survival rate was 60%. Overall satisfaction rate of patients was 93% (43/46).Conclusions:Compartment-based targeted fat grafting is an accurate injection method, which meets the requirement of physiological augmentation. The trajectory of targeted fat grafting will further improve the efficacy and safety of injection.

Panax notoginseng saponins(PNSs)as the extracted bioactive components of Panax notoginseng,have a long history of application in prevention of thrombosis.PNSs down-regulate the expression of inflammatory factors,promoting endothelial cell growth,up-regulating the expression of anticoagulants and vasodilators,and regulating endothelial cell function;With multiple targets at which PNSs inhibit platelet adhesion,release,and aggregation;PNSs maintain the activity of the fibrinolytic system by regulating the dynamic balance of tissue type plasminogen activators and inhibitors;PNSs reduce blood viscosity,improve red blood cell indicators,and inhibit thrombosis through multiple pathways.

To understand the current situation, progress, main contents, and the relevant assessment tools of family care in palliative for pediatric oncology patients, this paper reviewed the relevant literature on family care in palliative for pediatric oncology patients and its assessment tools at home and abroad. Taking family care in palliative care as the starting point, this paper discussed the effect of effective family care on improving the treatment outcome, quality of life, prognosis of pediatric oncology patients and the psychological problems of their families, and to provide a basis for continuing to improve the hospital-family-community care model for pediatric oncology patients, bringing into play the active role of family in palliative care, and promoting the continued development of family care for pediatric oncology patients.

Objective:The purpose of this study is to establish the trajectory of targeted grafting for facial fat compartment based on anatomical research, and then bring it to clinical practice.Methods:The boundary of fat compartment and the relationship of adjacent vessel and nerve were clarified through autopsy. The recommended injection points and trajectory for targeted fat grafting were established on the anatomical findings. Retrospective clinical data of facial rejuvenation of 46 patients through targeted fat grafting were collected from June 2017 to June 2019 in Shanghai Ninth People’s Hospital. The result of 3D scanning were analyzed to evaluate the survival rate of fat grafts.Results:There were subcutaneous superficial fat compartments in the frontal region, and there were both deep and superficial fat compartments in the temporal and middle face. According to the anatomical characteristics, a targeted fat grafting technique was established with the frontal hairline and the oral commissure corner mucosa as the entry points. In the clinical study, 46 patients were evaluated by 3D scanning 6 months after the last fat grafting. The amount of fat grafts in the temporal region was (17.84±8.47) ml and (11.2±2.44) ml was left after operation, and the survival rate was 63%. The amount of fat grafts in mid-face was (26.81±10.36) ml and (16.09±4.48) ml was left after operation, and the survival rate was 60%. Overall satisfaction rate of patients was 93% (43/46).Conclusions:Compartment-based targeted fat grafting is an accurate injection method, which meets the requirement of physiological augmentation. The trajectory of targeted fat grafting will further improve the efficacy and safety of injection.