Extensive epigenetic reprogramming involves in memory CD8+ T-cell differentiation. The elaborate epigenetic rewiring underlying the heterogeneous functional states of CD8+ T cells remains hidden. Here, we profile single-cell chromatin accessibility and map enhancer-promoter interactomes to characterize the differentiation trajectory of memory CD8+ T cells. We reveal that under distinct epigenetic regulations, the early activated CD8+ T cells divergently originated for short-lived effector and memory precursor effector cells. We also uncover a defined epigenetic rewiring leading to the conversion from effector memory to central memory cells during memory formation. Additionally, we illustrate chromatin regulatory mechanisms underlying long-lasting versus transient transcription regulation during memory differentiation. Finally, we confirm the essential roles of Sox4 and Nrf2 in developing memory precursor effector and effector memory cells, respectively, and validate cell state-specific enhancers in regulating Il7r using CRISPR-Cas9. Our data pave the way for understanding the mechanism underlying epigenetic memory formation in CD8+ T-cell differentiation.
CD8-Positive T-Lymphocytes/metabolism* ; Cell Differentiation ; Chromatin/immunology* ; Animals ; Mice ; Immunologic Memory ; Epigenesis, Genetic ; SOXC Transcription Factors/immunology* ; NF-E2-Related Factor 2/immunology* ; Mice, Inbred C57BL ; Gene Regulatory Networks ; Enhancer Elements, Genetic

Objective To observe the clinical efficacy of oral use combined with targeted transdermal delivery of Osteoking in treating knee osteoarthritis(KOA)of cold-damp blockage type.Methods A total of 120 patients with KOA of cold-damp blockage type who admitted to Hunan Provincial Hospital of Integrated Traditional Chinese And West Medicine from September 2022 to September 2023 were randomly divided into the control group,oral use group,transdermal delivery group and combination group according to the random number table method,with 30 cases in each group.The control group was treated with Celecoxib Capsule orally,the oral use group was treated with Osteoking orally,the transdermal delivery group was treated with Osteoking by targeted transdermal delivery,and the combination group was treated with oral use combined with targeted transdermal delivery of Osteoking.One course of treatment covered 12 days,and all of the four groups were treated for two continuous courses.Before and after treatment,the scores of traditional Chinese medicine(TCM)syndrome,Visual Analogue Scale(VAS)for pain,Western Ontario and McMaster Universities Osteoarthritis Index(WOMAC)for joint function,36-Item Short Form Health Survey(SF-36)for quality of life(QOL),as well as the levels of serum interleukin 1β(IL-1β),interleukin 6(IL-6),and tumor necrosis factor α(TNF-α)were observed in the patients of each group.After treatment,the clinical efficacy and safety of patients in each group were evaluated.Results(1)After two courses of treatment,the total effective rate of the combination group was 96.67%(29/30),which was higher than those of the control group[73.33%(22/30)],oral use group and transdermal delivery group[both being 70.00%(21/30)],and the difference was statistically significant(P<0.05).However,no statistically significant difference of the total effective rate was presented among the control group,oral use group,and transdermal delivery group(P>0.05).(2)After treatment,the scores of TCM syndrome,VAS for pain and WOMAC for joint function in the four groups were decreased compared with those before treatment(P<0.01).The intergroup comparison showed that the scores of TCM syndrome,VAS for pain and WOMAC in the combination group were lower than those in the other three groups(P<0.05),while no statistically significant difference of the scores of TCM syndrome,VAS for pain and WOMAC were presented among the control group,oral use group,and transdermal delivery group(P>0.05).(3)After treatment,the QOL scores of the eight dimensions of SF-36 such as physical functioning(PF),bodily pain(BP),general health(GH),vitality(VT),role-physical(RP),social functioning(SF),role-emotional(RE),and mental health(MH)in the four groups were significantly increased compared with those before treatment(P<0.01).The intergroup comparison showed that the scores of each dimension of SF-36 in the combination group were significantly higher than those in the other three groups(P<0.05),while no statistically significant difference of the dimension score of SF-36 was presented among the control group,oral use group,and transdermal delivery group(P>0.05).(4)After treatment,the levels of serum inflammatory factors of IL-1β,IL-6,and TNF-α in the four groups were decreased compared with those before treatment(P<0.01).The intergroup comparison showed that the decrease of serum levels of inflammatory factors in the combination group were significantly superior to those in the other three groups(P<0.05),and the decrease in the control group and oral use group were all superior to those in the transdermal delivery group(P<0.05),while the difference between the control group and oral use group was not statistically significant(P>0.05).(5)During the trial,no serious adverse reactions were found in the patients of four groups,which is of high safety.Conclusion Oral use combined with targeted transdermal delivery of Osteoking is effective on improving the joint pain,joint function and QOL of patients with KOA of cold-damp blockage type,and can speed up the rehabilitation of patients.Its therapeutic mechanism may be related to the decrease of the expression level of inflammatory factors.

BACKGROUND:Abnormal extracellular matrix accumulation and excessive proliferation of fibroblasts are the main manifestations of pathological scars.Excessive proliferation of fibroblasts leads to the production of large amounts of collagen-based extracellular matrix.Therefore,to investigate the role of fibroblast fibrosis in the formation of pathological scar will provide a new idea for revealing the mechanism of pathological scar and biological therapy. OBJECTIVE:To investigate the effect of RAS-selective lethal small molecule 3(RSL3)on the fibrosis of human pathological scar fibroblasts. METHODS:Then cases of pathological scar tissue and normal skin tissue samples from the same individuals,provided by the Department of Burn Plastic Surgery,General Hospital of Ningxia Medical University,were collected.Fibroblasts of human pathological scar and human normal skin were extracted and used in the following experiments.The general condition of the pathological scar tissue and the normal skin tissue was detected by hematoxylin-eosin staining.The appearance of fibroblasts from pathological scar and normal skin were observed by inverted microscope.The fibroblasts were verified by immunofluorescence assay.The cells were treated with different concentrations of RSL3(1,3,5,7,9,11,13 μmol/L).The inhibitory concentration of RSL3 on fibroblasts was detected by cell counting kit-8.Control group(without treatment)and RSL3 intervention group(treated with 7 μmol/L RSL3 for 24 hours)were set up.The mRNA and protein expressions of glutathione peroxidase 4,type Ⅰ collagen,type Ⅲ collagen and α-smooth muscle actin were detected by Qrt-PCR and western blot,respectively.Level of malondialdehyde in cells was detected.The residual scratch area was measured by cell scratch test after 24 hours to calculate the percentage of residual scratch area. RESULTS AND CONCLUSION:The expression of glutathione peroxidase 4 in the pathological scar group was higher than that in the normal skin group(Mrna:t=3.252,P<0.01;protein:t=5.075,P<0.01).The expression of glutathione peroxidase 4 in the pathological scar fibroblast group was higher than that in the normal skin fibroblast group(Mrna:t=10.32,P<0.01;protein:t=26.22,P<0.01).Compared with the control group,the expression of glutathione peroxidase 4 was decreased(Mrna:t=2.798,P<0.05;protein:t=4.643,P<0.01),the content of malondialdehyde was increased(t=2.917,P<0.05),the expression of type Ⅰ collagen(Mrna:t=15.84,P<0.01;protein:t=4.610,P<0.01),type Ⅲ collagen(Mrna:t=28.86,P<0.01;protein:t=7.713,P<0.01)and α-smooth muscle actin(Mrna:t=2.671,P<0.05;protein:t=7.417,P<0.01)were decreased in the RSL3 intervention group.Compared with the control group,the migration ability was weakened in the RSL3 intervention group(t=14.06,P<0.01).To conclude,RSL3 can inhibit the expression of glutathione peroxidase 4 and then inhibit the ability of fibrosis and migration of pathological scar fibroblasts.

Multiple primary lung cancer(MPLC)refers to patients with two or more primary lesions of lung cancer.It can be divided into synchronous MPLC(sMPLC)and metachronous MPLC(mMPLC)based on the timing of occurrence.In recent years,the detection rate of MPLC has gradually increased.However,considerable controversy exists in distinguishing MPLC from intrapulmonary metastasis(IM),especially when the histopathological types are identical.Given the significant differences in treatment strategies and prognosis in clinical practice currently,accurate diagnosis of MPLC is cru-cial for personalized precision therapy.Molecular genetics and sequencing technologies offer effective strategies for assessing the clonal origin of tumors.There have been reports of coexisting mutations in the epidermal growth factor receptor(EGFR)and anaplastic lymphoma kinase(ALK)fusion genes in non-small cell lung cancer,but case of EGFR mutation following an ALK mutation has not been mentioned.This article accurately diagnoses and retrospectively analyzes the clinical data of a case of ALK mutant adenocarcinoma in a male patient who developed an EGFR mutation with multiple metastases four years after surgery,and reviews the relevant literature.This paper aims to deepen the understanding of mMPLC and provide clinical refer-ences for the diagnosis and treatment of such patients.

Objective:To investigate the clinical outcome of biplanar botulinum toxin type A injection in alleviating platysmal bands.Methods:From November 2022 to May 2023, the clinical data of patients with platysmal bands treated by botulinum toxin type A injection in Department of Face and Neck Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, and Department of Plastic Surgery, Chengdu Badachu Cosmetic Hospital were retrospectively analyzed. The platysmal bands were marked, while patients were grinning, before injection. Using a 13 mm 30 G needle, 20 U/ml botulinum toxin was injected into the muscle layer along the bands from the clavicle direction. The dose was 1 U at a single point every 1.5 cm. Using a 3 mm 30 G needle, 10 U/ml botulinum toxin was injected into the deep surface of dermis along the bands with a single point dose of 0.5 U. Effects were evaluated by overall subjective satisfaction of patients, which were categorized into 4 grades: very satisfied, satisfied, dissatisfied, very dissatisfied. In addition, accessment by a third-party physician with global aesthetic improvement scale (GAIS) (1-5 points, the lower the score, the more significant the improvement is) and Geister platysmal band scale (0-4 points, the higher the score, the more severe the platysmal band is). Normal distribution data was represented by Mean±SD.Results:A total of 19 patients were included, including 3 males and 16 females, with the average age of 36.1 years. After a mean follow-up of 1.3 months (1-5 months), the overall subjective satisfaction was 100%(19/19). The GAIS score of third-party physicians was 1.12±0.33. 100%(19/19) of patients received a rating over moderate improvement(significant improvement in 17 cases and moderate improvement in 2 cases). The Geister platysmal band score decreased from preoperative 3.65 ± 0.33 to postoperative 0.76 ± 0.44. No serious complications were found except 5 cases of local congestion and 2 cases of injection pain, which were relieved in 1 week and 3 hours respectively. 2 cases felt mild neck weakness, but neck activity was not affected. The adverse symptoms all completely resolved spontaneously within 4 weeks. All patients have no mouth deviation, difficulty speaking, facial paralysis, allergies, or other noticeable complications.Conclusion:The injection of botulinum toxin type A in dual-plane of platysmal intramuscular layer and deep intradermal layer can effectively alleviate platysmal bands and achieve neck rejuvenation.

Objective:To investigate the clinical outcome of biplanar botulinum toxin type A injection in alleviating platysmal bands.Methods:From November 2022 to May 2023, the clinical data of patients with platysmal bands treated by botulinum toxin type A injection in Department of Face and Neck Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, and Department of Plastic Surgery, Chengdu Badachu Cosmetic Hospital were retrospectively analyzed. The platysmal bands were marked, while patients were grinning, before injection. Using a 13 mm 30 G needle, 20 U/ml botulinum toxin was injected into the muscle layer along the bands from the clavicle direction. The dose was 1 U at a single point every 1.5 cm. Using a 3 mm 30 G needle, 10 U/ml botulinum toxin was injected into the deep surface of dermis along the bands with a single point dose of 0.5 U. Effects were evaluated by overall subjective satisfaction of patients, which were categorized into 4 grades: very satisfied, satisfied, dissatisfied, very dissatisfied. In addition, accessment by a third-party physician with global aesthetic improvement scale (GAIS) (1-5 points, the lower the score, the more significant the improvement is) and Geister platysmal band scale (0-4 points, the higher the score, the more severe the platysmal band is). Normal distribution data was represented by Mean±SD.Results:A total of 19 patients were included, including 3 males and 16 females, with the average age of 36.1 years. After a mean follow-up of 1.3 months (1-5 months), the overall subjective satisfaction was 100%(19/19). The GAIS score of third-party physicians was 1.12±0.33. 100%(19/19) of patients received a rating over moderate improvement(significant improvement in 17 cases and moderate improvement in 2 cases). The Geister platysmal band score decreased from preoperative 3.65 ± 0.33 to postoperative 0.76 ± 0.44. No serious complications were found except 5 cases of local congestion and 2 cases of injection pain, which were relieved in 1 week and 3 hours respectively. 2 cases felt mild neck weakness, but neck activity was not affected. The adverse symptoms all completely resolved spontaneously within 4 weeks. All patients have no mouth deviation, difficulty speaking, facial paralysis, allergies, or other noticeable complications.Conclusion:The injection of botulinum toxin type A in dual-plane of platysmal intramuscular layer and deep intradermal layer can effectively alleviate platysmal bands and achieve neck rejuvenation.

Objective:To investigate the clinical characteristics and potential predisposing factors of the external cervical resorption(ECR).Methods:22 ECR cases with 38 affected teeth from 2016 to 2022 were retrospectively reviwed.Descriptive analysis combined with single factor analysis was used to study the clinical characteristics and influencing factors of ECR.Results:Maxillary anterior teeth(34.2％)were the most affected by ECR.Univariate analysis showed that ECR was more commonly noted in teeth without percussion pain and palpation pain,the probing depth of the periodontal pocket was greater than 3mm,with pulp activity reaction,without forma-tion of abscess and/or sinus tract,and without periapical lesions.There were statistically significant differences in percussion tender-ness,palpation tenderness and probing depth among the different Heithersary stages(P＜0.05).In the advanced cases,deep periodon-tal pockets and abscess formation were observed.The most common related dental factors of ECR were orthodontic treatment(15.87％)and dental traumatic injury(28.57％).Conclusion:ECR affected teeth often lack of clinical signs and symptoms.Radiology is the key to early diagnosis.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

The clinical and genetic characteristics of the family reported with neurodevelopmental syndrome caused by the SPTBN1 gene mutation were analyzed for clinical diagnosis.The proband was a boy, 2 years and 3 months old, admitted to the Department of Neurology, Guangzhou Women and Children′s Medical Center in June 2022 with comprehensive developmental delays as the main manifestation.The boy was backward in development since childhood.He was able to raise his head at the age of 3 months and sit alone at the age of 11 months.He could stand up with support but was unable to climb.He occasionally spoke polysyllabic words.The proband′s elder brother, 3 years and 1 month old, was able to walk at the age of 1 year and 6 months, and could speak " Mom and Dad" consciously and understand some instructions.He liked to play with other children.The mother of the proband was mentally retarded, while the father and grandparents of the proband had no symptoms.The proband was found to have a heterozygous mutation of the SPTBN1 gene (NM_003128.3), c.811G>A (p.Val271Met).The proband′s mother and elder brother also had a heterozygous mutation, which, however, was not detected in the proband′s father.The neurodevelopmental syndrome caused by the SPTBN1 gene mutation is rare in China, which can be manifested as language and motor delays and intellectual disabilities from early childhood, and individuals with the same genetic variation show different clinical phenotypes.