Objective:To observe the effects of kidney-tonifying and mind-calming acupuncture therapy on sleep,mood,sex hormone levels,and traditional Chinese medicine(TCM)symptoms in patients with perimenopausal insomnia(PMI). Methods:A total of 90 patients with PMI were randomly divided into a treatment group and a control group,with 45 cases in each group.Patients in the treatment group were treated with acupuncture at Shenshu(BL23),Taixi(KI3),Baihui(GV20),and Anmian(Extra).The control group was treated with sham acupuncture.Both groups were treated 3 times a week for 4 weeks.Pittsburgh sleep quality index(PSQI)and insomnia severity index(ISI)were used to evaluate the sleep quality of the subjects before treatment,after treatment,and 1 month after treatment(follow-up).Beck depression inventory(BDI)and Beck anxiety inventory(BAI)were used to evaluate the depression and anxiety of the subjects before treatment,after treatment,and at 1-month follow-up.The TCM symptom scale was used to evaluate the TCM symptoms of the subjects before treatment,after treatment,and 1 month after treatment.Serum levels of estradiol(E2),follicle-stimulating hormone(FSH),and luteinizing hormone(LH)were measured before and after treatment. Results:During the study,2 cases dropped out of the treatment group,and no cases dropped out of the control group.The PSQI scores of the treatment group were significantly lower after treatment and at 1-month follow-up compared with those before treatment(P<0.05),and the difference was statistically significant compared with that of the control group(P<0.05).In the control group,the PSQI score was significantly lower after treatment compared with before treatment(P<0.05),and the difference was not statistically significant at 1-month follow-up compared with before treatment(P>0.05).Compared with the pre-treatment,the ISI,BDI,BAI,and TCM symptom scale scores of the treatment group were lower after treatment and at 1-month follow-up(P<0.05),and the differences with the control group at the same time point were statistically significant(P<0.05).The differences in ISI,BDI,BAI,and TCM symptom scale scores of the control group before treatment,after treatment,and at 1-month follow-up were not statistically significant(P>0.05).After treatment,the serum E2 level in the treatment group was significantly higher than that before treatment(P<0.05),and the difference with the control group was statistically significant(P<0.05).The difference in the serum E2 level before and after treatment in the control group was not statistically significant(P>0.05).The differences in the serum FSH and LH levels between before and after treatment were not statistically significant in either group of subjects(P>0.05). Conclusion:Kidney-tonifying and mind-calming acupuncture therapy can improve sleep quality,relieve anxiety and depression,delay the decrease of serum E2 level,and improve related TCM symptoms in patients with PMI.

Objective The aim is to utilize CRISPR/Cas9 gene editing technology to construct Dmd gene mutant mice with a point mutation in exon 23 of the Dmd gene. Subsequently, the phenotypic changes of the mice in muscles and immune systems are analyzed and verified, providing an evaluation model for Duchenne muscular dystrophy and other related diseases.MethodsBased on the sequence characteristics of exon 23 of the Dmd gene, small guide RNA (sgRNA) was designed and synthesized. Cas9 mRNA, sgRNA fragments, and oligo donor DNA were microinjected into fertilized eggs of C57BL/6J mice. After transferring the fertilized eggs to surrogate mice, F0 generation mice were born. After mating with F0 generation mice, offspring mice were obtained, and Dmd gene positive mutant (Dmd^Mu/+) mice were obtained after genotype identification. Male hemizygous Dmd^Mu/+(Dmd^Mu/Y) mice were selected for phenotype validation. The body weight of live 3- and 9-month-old mice were recorded. Muscle tension was evaluated through the grid test. Hearts and semitendinosus muscles were collected, and the histopathological changes were observed using HE staining. Further, the expression of Dmd protein in muscle tissue of 9-month-old mice was analyzed by Western blotting.An acute inflammation model was established in Dmd^Mu/Y mice using lipopolysaccharide induction. Peripheral blood from the submandibular vein was collected, and the changes in the proportion of neutrophils and monocytes were detected by flow cytometry.Results The results of genome sequencing and Western blotting confirmed the successful construction of Dmd gene point mutant mice (Dmd^Mu/+ mice). Dmd protein expression was not detected in skeletal muscle and myocardium of Dmd^Mu/+ mice, and it was significantly reduced compared to wild-type C57BL/6J mice (P<0.05). Compared with wild-type mice of the same background, Dmd^Mu/Y mice at 3 and 9 months of age showed significant weight loss (P<0.01) and decreased muscle tension (P<0.05). 9-month-old Dmd^Mu/Y mice exhibited significant pathological changes in skeletal muscle and myocardium, including widening of intermuscular space. Under normal condition, compared with wild-type mice, the proportion of neutrophils and monocytes in the peripheral blood of 3-month-old Dmd^Mu/Y mice was significantly lower than that of wild-type mice (P<0.01). After lipopolysaccharide stimulation, the proportion of neutrophils in peripheral blood of 3-month-old Dmd^Mu/Y mice remained significantly lower compared to that of wild-type mice (P<0.01). The proportion of neutrophils in peripheral blood of 9-month-old Dmd^Mu/Y mice significantly decreased after lipopolysaccharide induction (P<0.01), with a trend of change observed in monocytes between groups.Conclusion The successful construction of the Dmd gene mutant mouse model has confirmed the vital function of Dmd gene in maintaining normal muscle tissue morphology and muscle tone. It preliminarily indicated that Dmd gene deletion could significantly reduce the proportion of neutrophils in peripheral blood, offering a new perspective for the study of immune system alterations in Duchenne muscular dystrophy patients.

OBJECTIVE To study the effects of transferrin-targeting peptide T7 （7pep） on intracellular transportation of polyethylene glycol-polycaprolactone （PEG-PCL） micelles in human cervical cancer HeLa cells. METHODS Using coumarin-6 （C6） as fluorescent indicator probe， both coumarin-6 （C6）-loaded PEG-PCL （PEG-PCL-C6） micelles and 7pep-modified PEG- PCL （7pep-PEG-PCL-C6） micelles were prepared by film-dispersion method. The particle size， polydispersity index and appearance morphology were compared between two types of micelles； the real-time uptake of two types of micelles by HeLa cells was compared， and the colocalization of two types of micelles with early endosomes （EE）， endocytic recycling compartments （ERC） and late endosomes （LE） after entry into the cells was observed. RESULTS The particle sizes of PEG-PCL-C6 and 7pep-PEG-PCL- C6 micelles were（75.0±2.3）and（82.0±1.5）nm； the polymer dispersity indexes were 0.17±0.20 and 0.17±0.32， respectively， with a regular spherical appearance. The colocalization results showed that entry speed and amount of 7pep-PEG-PCL-C6 micelles were significantly faster/more than those of PEG-PCL-C6 micelles. 7pep-PEG-PCL-C6 micelles entered EE faster than PEG-PCL-C6 micelles， while PEG-PCL-C6 micelles entered ERC at a faster rate than 7pep-PEG-PCL-C6 micelles， and both PEG-PCL-C6 micelles and 7pep-PEG-PCL-C6 micelles tended to accumulate gradually in LE； Pearson coefficient， signal overlap ratio， and colocalization ratio of 7pep-PEG-PCL-C6 micelles with LE were significantly lower 60 minutes after entering the cell than those 30 minutes after entering the cell （P＜0.05 or P＜0.01）. CONCLUSIONS Targeting 7pep modification can increase the entry speed and amount of PEG-PCL-C6 micelles， and also alter their intracellular transportation behavior.

The timely treatment of dental caries and pulp disease in primary teeth holds significant importance for maintaining children's oral health.Direct pulp capping(DPC)is a vital pulp treatment that involves covering the ex-posed pulp with bioactive materials to promote dentin bridge formation.DPC is commonly used in primary teeth with vi-tal pulp and mechanical pulp exposure not exceeding 1 mm.DPC offers advantages such as minimal invasiveness,com-fort,simplicity of operation and short chair-side time,making it suitable for pediatric dental clinical practice.Early stud-ies suggested negative treatment outcomes for DPC in primary teeth with carious pulp exposure.Over the years,there have been advancements in materials and technology demonstrating positive outcomes in the clinical research of prima-ry teeth with deep caries.However,due to the limited quality of related studies,DPC has not been widely recommended for the treatment of primary teeth with carious pulp exposure,and its widespread use needs further support by more high-quality evidence-based medical research.The success rate of DPC in primary teeth is influenced by factors including pulp status,clinical operations(such as isolation and caries removal),pulp capping material,cavity type,tooth position,coronal sealing,and dental fear.In clinical operation,dentists should accurately assess pulp status and minimize bacte-rial contamination.Mineral trioxide aggregate(MTA)is a DPC agent with relatively sufficient evidence and good thera-peutic effects,and the crown should be tightly sealed after pulp capping.Additionally,the effects of novel biocompati-ble materials such as iRoot BP Plus used in DPC of primary teeth,and the influence of other factors like hemostatic methods on the prognosis of affected teeth,need further exploration.

The timely treatment of dental caries and pulp disease in primary teeth holds significant importance for maintaining children's oral health.Direct pulp capping(DPC)is a vital pulp treatment that involves covering the ex-posed pulp with bioactive materials to promote dentin bridge formation.DPC is commonly used in primary teeth with vi-tal pulp and mechanical pulp exposure not exceeding 1 mm.DPC offers advantages such as minimal invasiveness,com-fort,simplicity of operation and short chair-side time,making it suitable for pediatric dental clinical practice.Early stud-ies suggested negative treatment outcomes for DPC in primary teeth with carious pulp exposure.Over the years,there have been advancements in materials and technology demonstrating positive outcomes in the clinical research of prima-ry teeth with deep caries.However,due to the limited quality of related studies,DPC has not been widely recommended for the treatment of primary teeth with carious pulp exposure,and its widespread use needs further support by more high-quality evidence-based medical research.The success rate of DPC in primary teeth is influenced by factors including pulp status,clinical operations(such as isolation and caries removal),pulp capping material,cavity type,tooth position,coronal sealing,and dental fear.In clinical operation,dentists should accurately assess pulp status and minimize bacte-rial contamination.Mineral trioxide aggregate(MTA)is a DPC agent with relatively sufficient evidence and good thera-peutic effects,and the crown should be tightly sealed after pulp capping.Additionally,the effects of novel biocompati-ble materials such as iRoot BP Plus used in DPC of primary teeth,and the influence of other factors like hemostatic methods on the prognosis of affected teeth,need further exploration.

The timely treatment of dental caries and pulp disease in primary teeth holds significant importance for maintaining children's oral health.Direct pulp capping(DPC)is a vital pulp treatment that involves covering the ex-posed pulp with bioactive materials to promote dentin bridge formation.DPC is commonly used in primary teeth with vi-tal pulp and mechanical pulp exposure not exceeding 1 mm.DPC offers advantages such as minimal invasiveness,com-fort,simplicity of operation and short chair-side time,making it suitable for pediatric dental clinical practice.Early stud-ies suggested negative treatment outcomes for DPC in primary teeth with carious pulp exposure.Over the years,there have been advancements in materials and technology demonstrating positive outcomes in the clinical research of prima-ry teeth with deep caries.However,due to the limited quality of related studies,DPC has not been widely recommended for the treatment of primary teeth with carious pulp exposure,and its widespread use needs further support by more high-quality evidence-based medical research.The success rate of DPC in primary teeth is influenced by factors including pulp status,clinical operations(such as isolation and caries removal),pulp capping material,cavity type,tooth position,coronal sealing,and dental fear.In clinical operation,dentists should accurately assess pulp status and minimize bacte-rial contamination.Mineral trioxide aggregate(MTA)is a DPC agent with relatively sufficient evidence and good thera-peutic effects,and the crown should be tightly sealed after pulp capping.Additionally,the effects of novel biocompati-ble materials such as iRoot BP Plus used in DPC of primary teeth,and the influence of other factors like hemostatic methods on the prognosis of affected teeth,need further exploration.

The timely treatment of dental caries and pulp disease in primary teeth holds significant importance for maintaining children's oral health.Direct pulp capping(DPC)is a vital pulp treatment that involves covering the ex-posed pulp with bioactive materials to promote dentin bridge formation.DPC is commonly used in primary teeth with vi-tal pulp and mechanical pulp exposure not exceeding 1 mm.DPC offers advantages such as minimal invasiveness,com-fort,simplicity of operation and short chair-side time,making it suitable for pediatric dental clinical practice.Early stud-ies suggested negative treatment outcomes for DPC in primary teeth with carious pulp exposure.Over the years,there have been advancements in materials and technology demonstrating positive outcomes in the clinical research of prima-ry teeth with deep caries.However,due to the limited quality of related studies,DPC has not been widely recommended for the treatment of primary teeth with carious pulp exposure,and its widespread use needs further support by more high-quality evidence-based medical research.The success rate of DPC in primary teeth is influenced by factors including pulp status,clinical operations(such as isolation and caries removal),pulp capping material,cavity type,tooth position,coronal sealing,and dental fear.In clinical operation,dentists should accurately assess pulp status and minimize bacte-rial contamination.Mineral trioxide aggregate(MTA)is a DPC agent with relatively sufficient evidence and good thera-peutic effects,and the crown should be tightly sealed after pulp capping.Additionally,the effects of novel biocompati-ble materials such as iRoot BP Plus used in DPC of primary teeth,and the influence of other factors like hemostatic methods on the prognosis of affected teeth,need further exploration.

The timely treatment of dental caries and pulp disease in primary teeth holds significant importance for maintaining children's oral health.Direct pulp capping(DPC)is a vital pulp treatment that involves covering the ex-posed pulp with bioactive materials to promote dentin bridge formation.DPC is commonly used in primary teeth with vi-tal pulp and mechanical pulp exposure not exceeding 1 mm.DPC offers advantages such as minimal invasiveness,com-fort,simplicity of operation and short chair-side time,making it suitable for pediatric dental clinical practice.Early stud-ies suggested negative treatment outcomes for DPC in primary teeth with carious pulp exposure.Over the years,there have been advancements in materials and technology demonstrating positive outcomes in the clinical research of prima-ry teeth with deep caries.However,due to the limited quality of related studies,DPC has not been widely recommended for the treatment of primary teeth with carious pulp exposure,and its widespread use needs further support by more high-quality evidence-based medical research.The success rate of DPC in primary teeth is influenced by factors including pulp status,clinical operations(such as isolation and caries removal),pulp capping material,cavity type,tooth position,coronal sealing,and dental fear.In clinical operation,dentists should accurately assess pulp status and minimize bacte-rial contamination.Mineral trioxide aggregate(MTA)is a DPC agent with relatively sufficient evidence and good thera-peutic effects,and the crown should be tightly sealed after pulp capping.Additionally,the effects of novel biocompati-ble materials such as iRoot BP Plus used in DPC of primary teeth,and the influence of other factors like hemostatic methods on the prognosis of affected teeth,need further exploration.

The timely treatment of dental caries and pulp disease in primary teeth holds significant importance for maintaining children's oral health.Direct pulp capping(DPC)is a vital pulp treatment that involves covering the ex-posed pulp with bioactive materials to promote dentin bridge formation.DPC is commonly used in primary teeth with vi-tal pulp and mechanical pulp exposure not exceeding 1 mm.DPC offers advantages such as minimal invasiveness,com-fort,simplicity of operation and short chair-side time,making it suitable for pediatric dental clinical practice.Early stud-ies suggested negative treatment outcomes for DPC in primary teeth with carious pulp exposure.Over the years,there have been advancements in materials and technology demonstrating positive outcomes in the clinical research of prima-ry teeth with deep caries.However,due to the limited quality of related studies,DPC has not been widely recommended for the treatment of primary teeth with carious pulp exposure,and its widespread use needs further support by more high-quality evidence-based medical research.The success rate of DPC in primary teeth is influenced by factors including pulp status,clinical operations(such as isolation and caries removal),pulp capping material,cavity type,tooth position,coronal sealing,and dental fear.In clinical operation,dentists should accurately assess pulp status and minimize bacte-rial contamination.Mineral trioxide aggregate(MTA)is a DPC agent with relatively sufficient evidence and good thera-peutic effects,and the crown should be tightly sealed after pulp capping.Additionally,the effects of novel biocompati-ble materials such as iRoot BP Plus used in DPC of primary teeth,and the influence of other factors like hemostatic methods on the prognosis of affected teeth,need further exploration.

The timely treatment of dental caries and pulp disease in primary teeth holds significant importance for maintaining children's oral health.Direct pulp capping(DPC)is a vital pulp treatment that involves covering the ex-posed pulp with bioactive materials to promote dentin bridge formation.DPC is commonly used in primary teeth with vi-tal pulp and mechanical pulp exposure not exceeding 1 mm.DPC offers advantages such as minimal invasiveness,com-fort,simplicity of operation and short chair-side time,making it suitable for pediatric dental clinical practice.Early stud-ies suggested negative treatment outcomes for DPC in primary teeth with carious pulp exposure.Over the years,there have been advancements in materials and technology demonstrating positive outcomes in the clinical research of prima-ry teeth with deep caries.However,due to the limited quality of related studies,DPC has not been widely recommended for the treatment of primary teeth with carious pulp exposure,and its widespread use needs further support by more high-quality evidence-based medical research.The success rate of DPC in primary teeth is influenced by factors including pulp status,clinical operations(such as isolation and caries removal),pulp capping material,cavity type,tooth position,coronal sealing,and dental fear.In clinical operation,dentists should accurately assess pulp status and minimize bacte-rial contamination.Mineral trioxide aggregate(MTA)is a DPC agent with relatively sufficient evidence and good thera-peutic effects,and the crown should be tightly sealed after pulp capping.Additionally,the effects of novel biocompati-ble materials such as iRoot BP Plus used in DPC of primary teeth,and the influence of other factors like hemostatic methods on the prognosis of affected teeth,need further exploration.