Periodontal bone defects, primarily caused by periodontitis, are highly prevalent in clinical settings and manifest as bone fenestration, dehiscence, or attachment loss, presenting a significant challenge to oral health. In regenerative medicine, harnessing developmental principles for tissue repair offers promising therapeutic potential. Of particular interest is the condensation of progenitor cells, an essential event in organogenesis that has inspired clinically effective cell aggregation approaches in dental regeneration. However, the precise cellular coordination mechanisms during condensation and regeneration remain elusive. Here, taking the tooth as a model organ, we employed single-cell RNA sequencing to dissect the cellular composition and heterogeneity of human dental follicle and dental papilla, revealing a distinct Platelet-derived growth factor receptor alpha (PDGFRA) mesenchymal stem/stromal cell (MSC) population with remarkable odontogenic potential. Interestingly, a reciprocal paracrine interaction between PDGFRA⁺ dental follicle stem cells (DFSCs) and CD31⁺ Endomucin⁺ endothelial cells (ECs) was mediated by Vascular endothelial growth factor A (VEGFA) and Platelet-derived growth factor subunit BB (PDGFBB). This crosstalk not only maintains the functionality of PDGFRA⁺ DFSCs but also drives specialized angiogenesis. In vivo periodontal bone regeneration experiments further reveal that communication between PDGFRA⁺ DFSC aggregates and recipient ECs is essential for effective angiogenic-osteogenic coupling and rapid tissue repair. Collectively, our results unravel the importance of MSC-EC crosstalk mediated by the VEGFA and PDGFBB-PDGFRA reciprocal signaling in orchestrating angiogenesis and osteogenesis. These findings not only establish a framework for deciphering and promoting periodontal bone regeneration in potential clinical applications but also offer insights for future therapeutic strategies in dental or broader regenerative medicine.
Receptor, Platelet-Derived Growth Factor alpha/metabolism* ; Humans ; Neovascularization, Physiologic/physiology* ; Dental Sac/cytology* ; Single-Cell Analysis ; Transcriptome ; Mesenchymal Stem Cells/metabolism* ; Bone Regeneration ; Animals ; Dental Papilla/cytology* ; Periodontium/physiology* ; Stem Cells/metabolism* ; Regeneration ; Angiogenesis

OBJECTIVE To observe the effect of electroacupuncture(EA)preconditioning at Tianshu(ST25)and Neiguan(PC6)on cardiac function and UCP1/BMP3b signaling pathway in brown adipose tissue(BAT)in mice with acute myocardial infarc-tion(AMI),so as to explore the potential mechanism of EA at different acupoints in improving myocardial infarction.METHODS Healthy adult mice and BAT excision mice were selected as the research objects,and they were divided into sham operation group,model group,PC6 group,and ST25 group.After one week of adaptive feeding,the mice in the intervention group were pretreated with bilateral EA at PC6 or ST25 for 20 minutes,respectively,and the AMI model was established by ligating the left anterior descending branch of the coronary artery.In the BAT resection group,BAT resection of the scapular region was performed before EA,and the rest of the intervention remained the same as before.Echocardiography was used to detect the changes in cardiac function.TTC staining was used to observe the myocardial infarct size.ELISA was used to detect the serum levels of cTnT and BMP3b in each group.The qPCR was used to detect the relative expression of β3-AR,UCP1 and BMP3b mRNA in mouse BAT.The protein expression of BMP3b in BAT and p-Smad1/5 in the heart were detected by Western blot.RESULTS Compared with the sham operation group,the left ven-tricular EF and FS of the model group mice were decreased(P<0.001),the white infarct area was increased(P<0.001),the cTnT level in serum was increased(P<0.001),the mRNA expression levels of β3-AR,UCP1,and BMP3b in BAT were increased(P<0.05,P<0.01),and the protein expression of BMP3b was increased(P<0.01).The BMP3b content in serum was increased(P<0.001),and the protein expression of p-Smad1/5 in the heart was increased(P<0.01).Compared with the model group,the left ventricular EF and FS of mice in the ST25 and PC6 group were increased(P<0.001),the area of white infarction was reduced(P<0.001),and the cTnT level in serum was decreased(P<0.05,P<0.01).The mRNA contents of β3-AR,UCP1,and BMP3b in BAT of the ST25 group were significantly increased(P<0.05,P<0.01,P<0.001),the protein expression of BMP3b was increased(P<0.01),and the protein expression of p-Smad1/5 in the heart was increased(P<0.01),while there was no significant change in the PC6 group.After BAT resection,compared with the model group,the left ventricular EF and FS of the mice in the PC6 group were increased(P<0.001),the area of white infarction was reduced(P<0.001),and the cTnT in serum was decreased(P<0.001),while there was no significant change in the ST25 group;there was no significant change in the protein expression of p-Smad1/5 in the heart and the BMP3b content in the serum of the PC6 and ST25 groups.CONCLUSION EA pretreatment at either ST25 or PC6 acupoints can produce myocardial protective effects.The protective effect of ST25 may be through influencing the UCP1/BMP3b signaling path-way in BAT,while PC6 does not depend on this pathway.

OBJECTIVE To observe the effect of electroacupuncture(EA)preconditioning at Tianshu(ST25)and Neiguan(PC6)on cardiac function and UCP1/BMP3b signaling pathway in brown adipose tissue(BAT)in mice with acute myocardial infarc-tion(AMI),so as to explore the potential mechanism of EA at different acupoints in improving myocardial infarction.METHODS Healthy adult mice and BAT excision mice were selected as the research objects,and they were divided into sham operation group,model group,PC6 group,and ST25 group.After one week of adaptive feeding,the mice in the intervention group were pretreated with bilateral EA at PC6 or ST25 for 20 minutes,respectively,and the AMI model was established by ligating the left anterior descending branch of the coronary artery.In the BAT resection group,BAT resection of the scapular region was performed before EA,and the rest of the intervention remained the same as before.Echocardiography was used to detect the changes in cardiac function.TTC staining was used to observe the myocardial infarct size.ELISA was used to detect the serum levels of cTnT and BMP3b in each group.The qPCR was used to detect the relative expression of β3-AR,UCP1 and BMP3b mRNA in mouse BAT.The protein expression of BMP3b in BAT and p-Smad1/5 in the heart were detected by Western blot.RESULTS Compared with the sham operation group,the left ven-tricular EF and FS of the model group mice were decreased(P<0.001),the white infarct area was increased(P<0.001),the cTnT level in serum was increased(P<0.001),the mRNA expression levels of β3-AR,UCP1,and BMP3b in BAT were increased(P<0.05,P<0.01),and the protein expression of BMP3b was increased(P<0.01).The BMP3b content in serum was increased(P<0.001),and the protein expression of p-Smad1/5 in the heart was increased(P<0.01).Compared with the model group,the left ventricular EF and FS of mice in the ST25 and PC6 group were increased(P<0.001),the area of white infarction was reduced(P<0.001),and the cTnT level in serum was decreased(P<0.05,P<0.01).The mRNA contents of β3-AR,UCP1,and BMP3b in BAT of the ST25 group were significantly increased(P<0.05,P<0.01,P<0.001),the protein expression of BMP3b was increased(P<0.01),and the protein expression of p-Smad1/5 in the heart was increased(P<0.01),while there was no significant change in the PC6 group.After BAT resection,compared with the model group,the left ventricular EF and FS of the mice in the PC6 group were increased(P<0.001),the area of white infarction was reduced(P<0.001),and the cTnT in serum was decreased(P<0.001),while there was no significant change in the ST25 group;there was no significant change in the protein expression of p-Smad1/5 in the heart and the BMP3b content in the serum of the PC6 and ST25 groups.CONCLUSION EA pretreatment at either ST25 or PC6 acupoints can produce myocardial protective effects.The protective effect of ST25 may be through influencing the UCP1/BMP3b signaling path-way in BAT,while PC6 does not depend on this pathway.

Objective: To observe the effects of electroacupuncture (EA) at Neiguan (PC6) on arrhythmia during acute myocardial ischemia-reperfusion and the expression of connexin 43 (Cx43) in rats. Methods: A total of 40 Sprague-Dawley male rats were used. Ten rats were randomly selected as the blank group, and the remaining 30 rats were randomly divided into a model group and an EA group, with 15 rats in each group. Before modeling, rats in the EA group received one session of EA intervention at bilateral Neiguan (PC6) for 30 min; the other groups were treated with the same grasping and anesthesia for 30 min without intervention. PowerLab physiological recorder was used to record electrocardiograph within 30 min of infarction. After the experiment, cardiac tissue and serum were collected from rats. Hematoxylin-eosin (HE) staining was used to observe the morphological changes of myocardial tissue in the ventricular infarction area of rats in each group. The expression of Cx43 protein in the myocardium of each group was detected by Western blotting (WB). Enzyme-linked immunosorbent assay (ELISA) was used to determine the activity of Na+-K+-ATPase in myocardial tissue and the serum content of endogenous digitalis-like factor (EDLF) in rats. Results: There was no statistical difference in arrhythmia score between the EA group and the model group, but the total duration and average duration of arrhythmia in the EA group were decreased (P<0.01). HE staining showed that compared with the blank group, myocardial cells in the model group were disorganized and seriously damaged. The pathological changes in the EA group were similar to those in the model group, but the damage was relatively minor. The results of WB showed that compared with the blank group, the Cx43 expression in myocardial tissue of the model group was decreased (P<0.01); compared with the model group, the Cx43 expression in the EA group was increased (P<0.01); compared with the blank group, the Na+-K+-ATPase activity in myocardial tissue of the model group was significantly decreased (P<0.01); compared with the model group, the Na+-K+-ATPase activity in the EA group was increased (P<0.01). ELISA results showed that compared with the blank group, the serum EDLF content in the model group was significantly increased (P<0.01); compared with the model group, the EDLF content in the EA group was decreased (P<0.01). Conclusion: EA at Neiguan (PC6) can delay and reduce the onset of arrhythmia during myocardial infarction in the rat model of myocardial ischemia-reperfusion. Its mechanism of action may be related to the regulation of the Cx43 expression in myocardial tissue, improvement of the activity of Na+-K+-ATPase in myocardial tissue, and increase in the content of serum EDLF.

Objective:To analyze the epidemiological features of patients with plasma cell leukemia (PCL) and calculate the prevalence of PCL in urban China in 2016.Methods:Calculation in this study was based on China's urban basic medical insurance from 23 provinces between January 1, 2016 and December 31, 2016. The identification of the patients with PCL was based on the disease names and codes in the claim data. Subgroup analyses were carried out by sex, region, and age. To test the robustness of the results, we performed sensitivity analyses. Age-adjusted prevalence was calculated, based on the 2010 Chinese census data.Results:The prevalence of PCL in urban China in 2016 was 0.11 per 100 000 population (95% CI 0.05-0.19) , and the male prevalence and female prevalence were 0.12 per 100 000 population (95% CI 0.06-0.21) and 0.10 per 100 000 population (95% CI 0.04-0.19) , respectively. The prevalence of PCL peaked at 70-79 years old. Sensitivity analyses proved the robustness of the primary result. The age-adjusted prevalence based on 2010 Chinese census data was 0.12 per 100 000 population (95% CI 0.11-0.13) . Conclusion:This study firstly analyzed the epidemiological characteristics of PCL in China, which can provide evidence for the research and policies regarding PCL.

OBJECTIVE: To investigate the difference of protective effect of electroacupuncture (EA) at different times pretreating myocardial ischemia-reperfusion injury (MIRI) and the protein expressions of ULK1 and Beclin-1 in myocardial tissue, so as to explore the relation of the difference and autophagic mechanism. METHODS: Sixty-three male SD rats were randomized into 7 groups, a sham group, a model group, an EA pretreating for 1 d (EA-1d) group, an EA pretreating for 2 d (EA-2d) group, an EA pretreating for 3 d (EA-3d) group, an EA pretreating for 4 d (EA-4d) group, an EA pretreating for 5 d (EA-5d) group, 9 rats in each group. All the rats in the pretreating groups were treated with EA 1-5 days before MIRI surgery. EA (2 Hz/100 Hz, 2 mA) was used at bilateral "Neiguan (PC 6)" for 20 min. All the rats except for those in the sham group was underwent left thoracotomy and ligation of the left anterior descending (LAD) coronary artery for 30 minutes followed by 4 hours of reperfusion to establish the MIRI model. The same operation was performed in the sham group except for the ligation of the LAD. Throughout the experiment, electrocardiogram was continuously monitored. The myocardial infarct sizes were assessed by Evans blue and triphenyltetrazolium chloride (TTC) staining. The serum concentrations of cardiac troponinⅠ(cTnⅠ) was detected by ELISA. The expressions of ULK1 and Beclin-1 in the heart tissues were analyze by Western blot. RESULTS: Compared with the sham group, the concentrations of cTnⅠ and the protein expressions of ULK1 and Beclin-1 increased in the model group (all <0.05). Compared with the model group, the infarct sizes decreased in the EA-1d, EA-2d, EA-3d, EA-4d and EA-5d groups (all <0.05), with less risk sizes in the EA-3d, EA-4d and EA-5d groups (all <0.05). The levels of cTnⅠin the EA-4d and EA-5d groups decreased ( both <0.05); the expressions of ULK1 protein decreased in the EA-1d, EA-2d, EA-3d, EA-4d and EA-5d groups (all <0.05); the expressions of Beclin-1 protein decreased in the EA-2d, EA-3d, EA-4d and EA-5d groups (all <0.05). The infarct sizes in the EA-4d and EA-5d groups were lower than that in the EA-1d group (both 0.05). The cTnⅠconcentration in the EA-4d group was less than that in the EA-1d group (<0.05). CONCLUSION Pretreatment with EA for 1-5 days can improve the infarct size in MIRI, with better effect of the pretreatment for 4-5 days. The cardioprotective effect may be related to the inhibition of autophagy. But the difference of the protective effects is not related to the protein expressions of ULK1 and Beclin-1.
Acupuncture Points ; Animals ; Autophagy ; Electroacupuncture ; Male ; Myocardial Ischemia ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To compare the effects of electroacupuncture (EA) at different time during reperfusion on the expression of autophagy-related protein Bcl-2 and Beclin1 in myocardial tissue in rats with myocardial ischemia reperfusion injury (MIRI), and to explore the autophagy-related mechanism of EA on protecting MIRI. METHODS: A total of 72 SD rats were randomly divided into a sham operation group, a model group, a RA group, a RB group, a RC group and a RD group, 12 rats in each group. Except the sham operation group, the rats in the remaining groups were treated with ligating the left anterior descending artery (LAD) for 30 minutes followed by reperfusion to establish the model of MIRI. The rats in the sham operation group were treated with crossing a line through the LAD. The rats in the model group did not receive treatment. The rats in the RA group, RB group, RC group and RD group were treated with EA at "Neiguan" (PC 6) for 20 min, starting at 0 h, 0.5 h, 1 h, 2 h after reperfusion. Evans Blue-TTC double-staining was employed to evaluate myocardial infarct size; the serum CK-MB was detected by ELISA and the expression of Bcl-2 and Beclin1 protein in myocardial tissue were detected by Western blot. RESULTS: Compared with the model group, the percentage of myocardial infarct size in the RB group, RC group and RD group was decreased significantly (all <0.05), and the reduction in the RB group was more significant than that in the RC group and RD group (both <0.05). Compared with the sham operation group, the expression of CK-MB and Beclin1 in the model group was significantly increased, but the expression of Bcl-2 was significantly decreased (all <0.01). Compared with the model group, the expression of CK-MB and Beclin1 was decreased significantly in RA group, RB group, RC group and RD group (all <0.05), but the expression of Bcl-2 was significantly increased (all <0.05). Compared with the RA group, the expression of CK-MB was decreased in the RB group and RC group (both <0.05) but the expression of Bcl-2 was increased (both <0.01); the expression was not significantly different from that in the RD group (>0.05); the increasing of Bcl-2 in the RB group was more significant than that in RC group (<0.05). The expression of Beclin 1 in the RB group was significantly lower than that in the RA group (<0.05), but there was no significant difference among other EA groups (all >0.05). CONCLUSION EA at different time during reperfusion could reduce myocardial infarct size in rats with MIRI, and EA at 0.5 h after reperfusion has best efficacy; this protective effect may be achieved by increasing Bcl-2 expression and reducing Beclin1 expression to inhibit overautophagy during reperfusion.
Animals ; Beclin-1 ; Electroacupuncture ; Humans ; Myocardial Ischemia ; Myocardial Reperfusion Injury ; Myocardium ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effects of preventative moxibustion on adenosine monophosphate-activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) in myocardial tissue in rats with exhaustive exercise, and to explore the action mechanism of preventative moxibustion for myocardial injury by exhaustive exercise.

METHODSEighteen SD male rats were randomly divided into a blank group, an exhaustive exercise group and a moxibustion group, 6 rats in each one. Rats in the blank group were treated with immobilization for 5 min per day, without any intervention. Rats in the exhaustive exercise group were treated with no intervention in the first 10 days. Rats in the moxibustion group were treated with moxibustion at "Zusanli" (ST 36) and "Guanyuan" (CV 4), 5 cones for each acupoint, for 10 days.On 11th day, rats in the exhaustive exercise group and moxibustion group were sacrificed to collect sample after exhaustive swimming, and time of exhaustive exercise was recorded. HE staining was used to observe the inflammatory changes of myocardial tissue; colorimetric method was used to measure lactic dehydrogenase (LDH), activity of superoxide dismutase (SOD) and malonaldehyde (MDA); immune suppression method was used to measure the content of creatine kinase isoenzyme (CK-MB) in serum; Elisa method was used to measure the content of troponinT (cTnT) in serum; western blot method was applied to measure the content of AMPKα2 and mTOR in myocardial tissue.

RESULTSThe time of exhaustive exercise was (4 831.17±689.88) s in the moxibustion group, which was longer than (3 509.50±1 232.49) s in the exhaustive exercise group (<0.05); HE staining indicated structure of the myocardium was clear and inflammatory cell infiltration was reduced in the moxibustion group. Compared with the blank group, the contents of serum LDH, CK-MB, cTnT were increased in the exhaustive exercise group (all<0.05); MDA content was increased and SOD activity was decreased (both<0.05); the AMPK α2 content in myocardial tissue was increased and the mTOR content was decreased (both<0.05). Compared with the exhaustive exercise group, the contents of serum LDH, CK-MB, cTnT were reduced in the moxibustion group (all<0.05); MDA content was decreased and SOD activity was increased (both<0.05); the AMPKα2 content in myocardial tissue was increased, and the mTOR content was decreased (both<0.05); AMPKα2 and mTOR were negatively correlated (=-0.764,<0.05).

CONCLUSIONSPreventative moxibustion is likely to regulate the expression of AMPK and mTOR to induce signaling pathway to recover myocardial injury by exhaustive exercise.