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 compare differences in the pharmacological activities of Angelica sinensis and its components on hematopoietic and laxative effects.Methods Kunming mice were randomly divided into eight groups,with 12 mice in each group consisting of equal numbers of males and females.These groups included a normal group,a model group,a positive group,a Angelica sinensis(AS)group,an Angelica sinensis water-soluble(AW)group,an Ethanol extract of Angelicae sinensis(AE)group,and an Angelica sinensis essential oil(AO)group.Except for the normal group,all other groups were established as blood deficiency constipation mouse models through subcutaneous injection of N-acetylphenylhydrazine combined with oral administration of loperamide hydrochloride.On the 7th day of modeling,each group received oral administration of the respective test substance once daily for three consecutive days.General condition and body weight changes of the mice were observed,peripheral blood cells were counted,stool morphology and fecal output were recorded,fecal moisture content and colonic tissue moisture content were determined,small intestine propulsion rate was assessed by a charcoal meal method,and serum levels of β-endorphin(β-EP),cholecystokinin octapeptide(CCK-8),substance P(SP),and vasoactive intestinal peptide(VIP)were determined by ELISA.Differences in the pharmacological activities of Angelica sinensis and its components on hematopoietic and laxative effects were analyzed.Results Compared with the normal group,model group mice showed significantly reduced white blood cell(WBC),red blood cell(RBC),hemoglobin(HGB),hematocrit(HCT),and platelet(PLT)counts,and body weight(P＜0.05 or P＜0.01).Additionally,fecal moisture content,colon moisture content,and small intestine propulsion rate were decreased(P＜0.05 or P＜0.01)and serum CCK-8 and SP levels were also lower(P＜0.01),while serum β-EP and VIP levels increased(P＜0.05).Compared with the model group,AS and AW groups had higher WBC,RBC,HGB,HCT,and PLT counts,defecation volume,fecal moisture content,and colon moisture content(P＜0.05 or P＜0.01).The AE group showed increased WBC,RBC,HGB,HCT,and PLT counts,and colon moisture content(P＜0.05 or P＜0.01),but defecation volume and fecal moisture content were not significantly altered.The AO group exhibited increased fecal moisture content,colon moisture content,and defecation volume(P＜0.05 or P＜0.01),but no significant changes in WBC,RBC,HGB,HCT,and PLT counts.The AE group showed no significant changes in defecation volume,fecal moisture content,and colon moisture content.The AS and AO groups had increased small intestine propulsion rates(P＜0.01),while there was no significant difference in small intestine propulsion rate between the AW and AE groups.The AS group had elevated serum CCK-8 and SP levels(P＜0.01)and decreased serum β-EP and VIP levels(P＜0.01).The AO group had increased serum CCK-8 and SP levels(P＜0.05),but no significant change in serum β-EP and VIP levels.The AW group had decreased serum VIP levels(P＜0.05),but no statistically significant difference in serum CCK-8 and SP levels.Compared with the AS group,the AW group had higher WBC,RBC,HGB,HCT,and PLT counts,while the AO and AE groups had lower levels of these parameters(P＜0.05).Both AW and AO groups had increased fecal moisture content(P＜0.05),and both AW and AE groups had increased colon moisture content(P＜0.05).AO,AE,and AW groups had elevated serum CCK-8 and SP levels and decreased serum β-EP and VIP levels(P＜0.05).In summary,the groups were ordered as follows:AE＞AO＞AS＞AW in terms of blood replenishment,AO＞AS＞AW＞AE in terms of promoting bowel movements,and AO＞AS＞AE＞AW in terms of intestinal motility.Conclusions Angelica sinensis and its components have varying degrees of blood replenishing and bowel-promoting activities.The AE component has strong blood replenishing activity,while the AO component has strong bowel-promoting and defecation-stimulating activity.These findings provide a reference for the development of traditional Chinese medicines based on Angelica sinensis components.

Objective To compare differences in the pharmacological activities of Angelica sinensis and its components on hematopoietic and laxative effects.Methods Kunming mice were randomly divided into eight groups,with 12 mice in each group consisting of equal numbers of males and females.These groups included a normal group,a model group,a positive group,a Angelica sinensis(AS)group,an Angelica sinensis water-soluble(AW)group,an Ethanol extract of Angelicae sinensis(AE)group,and an Angelica sinensis essential oil(AO)group.Except for the normal group,all other groups were established as blood deficiency constipation mouse models through subcutaneous injection of N-acetylphenylhydrazine combined with oral administration of loperamide hydrochloride.On the 7th day of modeling,each group received oral administration of the respective test substance once daily for three consecutive days.General condition and body weight changes of the mice were observed,peripheral blood cells were counted,stool morphology and fecal output were recorded,fecal moisture content and colonic tissue moisture content were determined,small intestine propulsion rate was assessed by a charcoal meal method,and serum levels of β-endorphin(β-EP),cholecystokinin octapeptide(CCK-8),substance P(SP),and vasoactive intestinal peptide(VIP)were determined by ELISA.Differences in the pharmacological activities of Angelica sinensis and its components on hematopoietic and laxative effects were analyzed.Results Compared with the normal group,model group mice showed significantly reduced white blood cell(WBC),red blood cell(RBC),hemoglobin(HGB),hematocrit(HCT),and platelet(PLT)counts,and body weight(P＜0.05 or P＜0.01).Additionally,fecal moisture content,colon moisture content,and small intestine propulsion rate were decreased(P＜0.05 or P＜0.01)and serum CCK-8 and SP levels were also lower(P＜0.01),while serum β-EP and VIP levels increased(P＜0.05).Compared with the model group,AS and AW groups had higher WBC,RBC,HGB,HCT,and PLT counts,defecation volume,fecal moisture content,and colon moisture content(P＜0.05 or P＜0.01).The AE group showed increased WBC,RBC,HGB,HCT,and PLT counts,and colon moisture content(P＜0.05 or P＜0.01),but defecation volume and fecal moisture content were not significantly altered.The AO group exhibited increased fecal moisture content,colon moisture content,and defecation volume(P＜0.05 or P＜0.01),but no significant changes in WBC,RBC,HGB,HCT,and PLT counts.The AE group showed no significant changes in defecation volume,fecal moisture content,and colon moisture content.The AS and AO groups had increased small intestine propulsion rates(P＜0.01),while there was no significant difference in small intestine propulsion rate between the AW and AE groups.The AS group had elevated serum CCK-8 and SP levels(P＜0.01)and decreased serum β-EP and VIP levels(P＜0.01).The AO group had increased serum CCK-8 and SP levels(P＜0.05),but no significant change in serum β-EP and VIP levels.The AW group had decreased serum VIP levels(P＜0.05),but no statistically significant difference in serum CCK-8 and SP levels.Compared with the AS group,the AW group had higher WBC,RBC,HGB,HCT,and PLT counts,while the AO and AE groups had lower levels of these parameters(P＜0.05).Both AW and AO groups had increased fecal moisture content(P＜0.05),and both AW and AE groups had increased colon moisture content(P＜0.05).AO,AE,and AW groups had elevated serum CCK-8 and SP levels and decreased serum β-EP and VIP levels(P＜0.05).In summary,the groups were ordered as follows:AE＞AO＞AS＞AW in terms of blood replenishment,AO＞AS＞AW＞AE in terms of promoting bowel movements,and AO＞AS＞AE＞AW in terms of intestinal motility.Conclusions Angelica sinensis and its components have varying degrees of blood replenishing and bowel-promoting activities.The AE component has strong blood replenishing activity,while the AO component has strong bowel-promoting and defecation-stimulating activity.These findings provide a reference for the development of traditional Chinese medicines based on Angelica sinensis components.

Objective: To propose updates and amendments for the nitroglycerin tablets quality in the Chinese pharmacopoeia 2020 due to the failure of effective separation of 4 known impurities and nonseparation of free nitrate ion and excipients.
Methods: Related substances were analyzed using gradient elution by HPLC, and free nitrate ion was determined on SAX column by different HPLC method.
Results: Using the improved method to test the related substances and free nitrate ion of nitroglycerin tablets,the content of the maximum individual impurity were not more than 0.5%, the total content was not more than 2.4% and the content of free nitrate ion was not more than 6.3%.
Conclusion: The improved method is accurate and feasible. It provided a reference for the updates and amendments of the quality standard of nitroglycerin tablets in the Chinese Pharmacopoeia 2020.

This paper reports a case of reparing the tissue defect after resection of a malignant tumor in the glabellar nasion with bilateral modified horizontal advancement flaps. The patient, a 20-year-old female, was admitted to the hospital because of a mass on the nasion between the eyebrows for more than 5 years, with enlargement and ulceration for 5 months. Surgical resection of the tumor was performed. Intraoperative rapid frozen sections showed that the tumor was basal cell carcinoma, and there were no cancer cells at the margins. Bilateral modified horizontal advancement flap repair was performed. At three months after the operation, the incision area healed well, the appearance and eyelid function were normal, and there was no sign of recurrence of basal cell carcinoma. Bilateral modified horizontal advancement flap was effective in repairing the tissue defect after resection of glabellar nasal root basal cell carcinoma.

Objective:To explore the surgical effects of continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty.Methods:A retrospective study was conducted on patients who underwent the operation of continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty at the Department of Plastic and Aesthetic Surgery of Qingdao Municipal Hospital from September 2021 to September 2023. Six points were marked on the skin surface of the designed double eyelid line on patient' s upper eyelid, 8 points were marked on the corresponding conjunctival surface of the designed double eyelid line. A 6-0 nylon thread attached with one needle on each end was used for suturing, with two ends running through and between the subcutaneous and subconjunctival layers alternatively, forming a " trapezoidal" shape of path. Finally, two ends exited at the same point on the most lateral side of the designed double eyelid line on skin surface and were tied together with the knot hidden within the orbicularis oculi muscle. The morphology of the double eyelids and postoperative complications were observed and evaluated after surgery, the follow-up period was at least 6 months. At the last time of follow-up, the satisfaction rates were evaluated by patients themselves. Four aspects of the surgical effects of the double eyelids were put into the evaluation system, which were radian, width, smoothness and symmetry, with score from 0-5 points for each. The total score was 0-20 points, of which 17-20 points indicated high satisfaction, score of 13-16 points indicated relative satisfaction, ≤ 12 points indicated dissatisfaction.Results:A total of 52 female patients were included, aged 18 to 29 years old, with an average age of 22.5 years. Among all patients, 33 cases had shallow double eyelid lines while 19 cases had single eyelids. All patients’ upper eyelids were relatively thin and without obvious looseness. The surgical procedures were all done smoothly and successfully, with an average operation time of (0.72±0.13) h. Immediately after surgery, the radian of the double eyelid lines on both sides looked smooth, the widths of the double eyelids were also basically symmetrical. During the follow-up period of 6-12 months postoperatively, the morphologies of the double eyelids were relatively natural and realistic, the double eyelid lines and widths remained deep and stable, and the incision scars were left unobvious for all patients. Besides, no complications such as ptosis, incomplete upper eyelid closure and ectropion were observed. Among the 52 patients, 45 cases were highly satisfied with the surgical result (17-20 points) , 7 were relatively satisfied (13-16 points) , while none of them required reoperation.Conclusions:The continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty has advantages of short operation time, good and stable surgical effects such as natural and realistic morphology of double eyelids, few complications, and high patients' satisfaction rate.

Objective:To explore the surgical effects of continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty.Methods:A retrospective study was conducted on patients who underwent the operation of continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty at the Department of Plastic and Aesthetic Surgery of Qingdao Municipal Hospital from September 2021 to September 2023. Six points were marked on the skin surface along the designed double eyelid line on patient’s upper eyelid, 8 points were marked on the corresponding conjunctival surface along the designed double eyelid line. A 6-0 nylon thread attached with one needle on each end was used for suturing, with two ends running through and between the subcutaneous and subconjunctival layers alternatively, forming a "trapezoidal" shape of path. Finally, two ends exited at the same point on the most lateral side of the designed double eyelid line on skin surface and were tied together with the knot hidden within the orbicularis oculi muscle. The morphology of the double eyelids and postoperative complications were observed and evaluated after surgery, the follow-up period was at least 6 months. At the last time of follow-up, the satisfaction rates were evaluated by patients themselves. Four aspects of the surgical effects of the double eyelids were put into the evaluation system, which were radian, width, smoothness and symmetry, with score from 0-5 points for each. The total score was 0-20 points, of which 17-20 points indicated high satisfaction, 13-16 points indicated relative satisfaction, ≤12 points indicated dissatisfaction.Results:A total of 52 female patients were included, aged 18 to 29 years old, with an average age of 22.5 years. Among all patients, 33 cases had shallow double eyelid lines while 19 cases had single eyelids. All patients’ upper eyelids were relatively thin and without obvious looseness. The surgical procedures were all done smoothly and successfully, with an average operation time of (0.72±0.13) h. Immediately after surgery, the radian of the double eyelid lines on both sides looked smooth, the widths of the double eyelids were also basically symmetrical. During the follow-up period of 6-12 months postoperatively, the morphologies of the double eyelids were relatively natural and realistic, the double eyelid lines and widths remained deep and stable, and the incision scars were unobvious for all patients. Besides, no complications such as ptosis, incomplete upper eyelid closure and ectropion were observed. Among the 52 patients, 45 cases were highly satisfied with the surgical result, 7 cases were relatively satisfied, while none of them required reoperation.Conclusion:The continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty has advantages of short operation time, good and stable surgical effects such as natural and realistic morphology of double eyelids, few complications, and high patients’ satisfaction rate.

This paper reports a case of reparing the tissue defect after resection of a malignant tumor in the glabellar nasion with bilateral modified horizontal advancement flaps. The patient, a 20-year-old female, was admitted to the hospital because of a mass on the nasion between the eyebrows for more than 5 years, with enlargement and ulceration for 5 months. Surgical resection of the tumor was performed. Intraoperative rapid frozen sections showed that the tumor was basal cell carcinoma, and there were no cancer cells at the margins. Bilateral modified horizontal advancement flap repair was performed. At three months after the operation, the incision area healed well, the appearance and eyelid function were normal, and there was no sign of recurrence of basal cell carcinoma. Bilateral modified horizontal advancement flap was effective in repairing the tissue defect after resection of glabellar nasal root basal cell carcinoma.

Objective:To explore the surgical effects of continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty.Methods:A retrospective study was conducted on patients who underwent the operation of continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty at the Department of Plastic and Aesthetic Surgery of Qingdao Municipal Hospital from September 2021 to September 2023. Six points were marked on the skin surface of the designed double eyelid line on patient' s upper eyelid, 8 points were marked on the corresponding conjunctival surface of the designed double eyelid line. A 6-0 nylon thread attached with one needle on each end was used for suturing, with two ends running through and between the subcutaneous and subconjunctival layers alternatively, forming a " trapezoidal" shape of path. Finally, two ends exited at the same point on the most lateral side of the designed double eyelid line on skin surface and were tied together with the knot hidden within the orbicularis oculi muscle. The morphology of the double eyelids and postoperative complications were observed and evaluated after surgery, the follow-up period was at least 6 months. At the last time of follow-up, the satisfaction rates were evaluated by patients themselves. Four aspects of the surgical effects of the double eyelids were put into the evaluation system, which were radian, width, smoothness and symmetry, with score from 0-5 points for each. The total score was 0-20 points, of which 17-20 points indicated high satisfaction, score of 13-16 points indicated relative satisfaction, ≤ 12 points indicated dissatisfaction.Results:A total of 52 female patients were included, aged 18 to 29 years old, with an average age of 22.5 years. Among all patients, 33 cases had shallow double eyelid lines while 19 cases had single eyelids. All patients’ upper eyelids were relatively thin and without obvious looseness. The surgical procedures were all done smoothly and successfully, with an average operation time of (0.72±0.13) h. Immediately after surgery, the radian of the double eyelid lines on both sides looked smooth, the widths of the double eyelids were also basically symmetrical. During the follow-up period of 6-12 months postoperatively, the morphologies of the double eyelids were relatively natural and realistic, the double eyelid lines and widths remained deep and stable, and the incision scars were left unobvious for all patients. Besides, no complications such as ptosis, incomplete upper eyelid closure and ectropion were observed. Among the 52 patients, 45 cases were highly satisfied with the surgical result (17-20 points) , 7 were relatively satisfied (13-16 points) , while none of them required reoperation.Conclusions:The continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty has advantages of short operation time, good and stable surgical effects such as natural and realistic morphology of double eyelids, few complications, and high patients' satisfaction rate.

Objective:To explore the surgical effects of continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty.Methods:A retrospective study was conducted on patients who underwent the operation of continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty at the Department of Plastic and Aesthetic Surgery of Qingdao Municipal Hospital from September 2021 to September 2023. Six points were marked on the skin surface along the designed double eyelid line on patient’s upper eyelid, 8 points were marked on the corresponding conjunctival surface along the designed double eyelid line. A 6-0 nylon thread attached with one needle on each end was used for suturing, with two ends running through and between the subcutaneous and subconjunctival layers alternatively, forming a "trapezoidal" shape of path. Finally, two ends exited at the same point on the most lateral side of the designed double eyelid line on skin surface and were tied together with the knot hidden within the orbicularis oculi muscle. The morphology of the double eyelids and postoperative complications were observed and evaluated after surgery, the follow-up period was at least 6 months. At the last time of follow-up, the satisfaction rates were evaluated by patients themselves. Four aspects of the surgical effects of the double eyelids were put into the evaluation system, which were radian, width, smoothness and symmetry, with score from 0-5 points for each. The total score was 0-20 points, of which 17-20 points indicated high satisfaction, 13-16 points indicated relative satisfaction, ≤12 points indicated dissatisfaction.Results:A total of 52 female patients were included, aged 18 to 29 years old, with an average age of 22.5 years. Among all patients, 33 cases had shallow double eyelid lines while 19 cases had single eyelids. All patients’ upper eyelids were relatively thin and without obvious looseness. The surgical procedures were all done smoothly and successfully, with an average operation time of (0.72±0.13) h. Immediately after surgery, the radian of the double eyelid lines on both sides looked smooth, the widths of the double eyelids were also basically symmetrical. During the follow-up period of 6-12 months postoperatively, the morphologies of the double eyelids were relatively natural and realistic, the double eyelid lines and widths remained deep and stable, and the incision scars were unobvious for all patients. Besides, no complications such as ptosis, incomplete upper eyelid closure and ectropion were observed. Among the 52 patients, 45 cases were highly satisfied with the surgical result, 7 cases were relatively satisfied, while none of them required reoperation.Conclusion:The continuous trapezoidal suture method between subcutaneous and subconjunctival tissue in buried-suture double-eyelid blepharoplasty has advantages of short operation time, good and stable surgical effects such as natural and realistic morphology of double eyelids, few complications, and high patients’ satisfaction rate.