Objective To investigate the effect of iRoot BP Plus on the repair of dental pulp cavity during pulp injury in rats,and to explore the possible role of iRoot BP Plus in the process of damage repair.Methods Bilateral maxillary first molars of 60 8-week-old SD rats were treated with direct pulp capping after pulp opening.They were divided into iRoot BP Plus group(BP group)and control group.After histological processing,the damage of dental pulp tissue and the calcification of pulp cavities were observed by microCT and HE staining.mRNA was extracted from rat dental pulp tissue,and the expression of inflammation and mineralization genes were detected by real-time quantitatitive PCR.Results The results of this study showed that the dental pulp tissue had different degrees of calcification at 1,4 and 7 days after direct pulp capping.There was no significant difference between the BP group and the control group on the 1st day after the operation,but on the 4th and 7th day after the operation,a regular calcified bridge was formed at the pulp hole in the BP group,and more pulp tissue was retained in the pulp cavity.In the control group,a large area of irregular calcified tissue was formed below the pulp hole.HE staining results showed that the pulp tissue had a heavy inflammatory response,and a large number of inflammatory cells infiltrated near the pulp orifice on the first day after the operation.On the 4th and 7th days after the opera-tion,mineralized tissue began to form near the pulp orifice in the BP group,and healthier pulp tissue was retained in the pulp cavity.The gene levels of IL-1β and TNF-α in the BP group were lower than those in the control group,and the gene expression levels of min-eralization indexes COL1 and DSPP were higher than those in the control group.The difference was statistically significant(P＜0.05).Conclusion Dental pulp injury can induce the immune defense response.If the inflammation is not well controlled,it will stimulate the calcification of dental pulp cavities.iRoot BP Plus can better regulate the dental pulp microenvironment,control the development of inflammation,reduce the formation of adverse calcified masses in the pulp cavity after pulp injury,and preserve more pulp tissue.

Aim To investigate the neuroprotective effect of Takeda G protein-coupled receptor-5(TGR5)activated by INT-777 on hypoxic-ischemic encephalop-athy(HIE)in neonatal rats.Methods Seven-day-old SD rats were randomly divided into the sham opera-tion group(Sham,S),model group(HIE,G),INT-777 low-dose(L),medium-dose(M),and high-dose(H)groups.The modified Rice-Vanucci method was used to construct the HIE model and Intranasal admin-istration 1 h after modeling.Short-term neurobehavioral tests were performed 48 h after modeling to evaluate the neurological function of neonatal rats,TTC staining was used to determine the volume of cerebral infarction,dry and wet specific gravity was used to determine the brain water content,ferrous ion kit was used to deter-mine the brain ferrous ion content,HE staining was used to observe the pathological damage of brain tis-sue,Nissl staining was used to observe the loss of Nissl substance,Transmission electron microscopy(TEM)was used to observe the mitochondrial morphological changes of cortical neurons,and Western blot was em-ployed to detect the expression of ferroptosis-related proteins TFR1 and GPX4.Results Compared with group S,group G had increased short-term neurobehav-ioral test consumption time,higher scores,increased cerebral infarct volume,brain water content,and brain ferrous iron content,significant brain tissue damage on the affected side,severe loss of Nissl substance,smaller neuronal mitochondria,decreased mitochondrial cris-tae,and increased expression of TFR1 and reduced ex-pression of GPX4.Compared with group G,the INT-777 administration group had a shorter consumption time for short-term neurobehavioral tests,lower scores,the cerebral infarction volume,brain water content,and brain ferrous ion content decreased,the brain tissue damage on the affected side was reduced,and there was insignificant loss of Nissl substance,larger neuronal mi-tochondrial volume,increased mitochondrial cristae,re-duced expression of TFR1,and increased expression of GPX4.Conclusions INT-777 agonist TGR5 has a protective effect against hypoxic-ischemic encephalopa-thy in neonatal rats,and its mechanism of action may be related to the inhibition of neuronal ferroptosis.

Aim To investigate the neuroprotective effect of Takeda G protein-coupled receptor-5(TGR5)activated by INT-777 on hypoxic-ischemic encephalop-athy(HIE)in neonatal rats.Methods Seven-day-old SD rats were randomly divided into the sham opera-tion group(Sham,S),model group(HIE,G),INT-777 low-dose(L),medium-dose(M),and high-dose(H)groups.The modified Rice-Vanucci method was used to construct the HIE model and Intranasal admin-istration 1 h after modeling.Short-term neurobehavioral tests were performed 48 h after modeling to evaluate the neurological function of neonatal rats,TTC staining was used to determine the volume of cerebral infarction,dry and wet specific gravity was used to determine the brain water content,ferrous ion kit was used to deter-mine the brain ferrous ion content,HE staining was used to observe the pathological damage of brain tis-sue,Nissl staining was used to observe the loss of Nissl substance,Transmission electron microscopy(TEM)was used to observe the mitochondrial morphological changes of cortical neurons,and Western blot was em-ployed to detect the expression of ferroptosis-related proteins TFR1 and GPX4.Results Compared with group S,group G had increased short-term neurobehav-ioral test consumption time,higher scores,increased cerebral infarct volume,brain water content,and brain ferrous iron content,significant brain tissue damage on the affected side,severe loss of Nissl substance,smaller neuronal mitochondria,decreased mitochondrial cris-tae,and increased expression of TFR1 and reduced ex-pression of GPX4.Compared with group G,the INT-777 administration group had a shorter consumption time for short-term neurobehavioral tests,lower scores,the cerebral infarction volume,brain water content,and brain ferrous ion content decreased,the brain tissue damage on the affected side was reduced,and there was insignificant loss of Nissl substance,larger neuronal mi-tochondrial volume,increased mitochondrial cristae,re-duced expression of TFR1,and increased expression of GPX4.Conclusions INT-777 agonist TGR5 has a protective effect against hypoxic-ischemic encephalopa-thy in neonatal rats,and its mechanism of action may be related to the inhibition of neuronal ferroptosis.

Objective By observing the regulatory effect of Strengthening Spleen and Eliminating Cancer Formula on N6-methyladenosine(m6A)methyltransferase,To explore the effect of Strengthening Spleen and Eliminating Cancer Formula on inhibiting the IRX5 m6A level in colorectal cancer(CRC),the regulatory effect on N6-methyladenosine(m6A)methyltransferase was observed.Methods Clinically,m6A hypermethylated genes in colorectal cancer was analyzed by m6A sequencing of pathological tissues from five CRC patients after radical surgery,looking for protein detection indexes for validation.23 BALB/c nude mice were selected and injected with HCT116 cells to establish a nude-mouse transplantation model of human colorectal cancer.They were divided into Model group,Western medicine group(5-fluorouracil group),Chinese medicine group(Jianpi Xiaoai Formula low-dose group,Jianpi Xiaoai Formula high-dose group),with 6 rats in each group,5 rats in control group.The tumor volume of all groups was compared.The overall methylation level of m6A was detected by colorimetric method.The protein expression levels of METTL3,METTL14,and WTAP,in tumor were detected by Western blot.The SRAMP website was used to predict the m6A sites of IRX5.HCT116 cells were treated with oe-NC,oe-METTL3,sh-NC,and sh-METTL3.The expression of IRX5 protein was detected by Western blot.HCT116 cell line was treated with Jianpi Xiaoai Formula drug-containing serum,and transfected with oe-METTL3 and oe-IRX5.The group was set as followed:control group,Jianpi Xiaoai Formula drug-containing serum group,Jianpi Xiaoai Formula drug-containing serum group+oe-NC,Jianpi Xiaoai Formula drug-containing serum group+oe-METTL3,Jianpi Xiaoai Formula drug-containing serum group+oe-IRX5,cell cloning experiment and Transwell experiment were performed to detect cell proliferation,migration and invasion ability of each group.The protein expression levels of METTL3 and IRX5 were detected by Western blot.Results The results of m6A sequencing of genes showed that the m6A methylation level increased in patients with CRC,and the m6A methylation levels of SOX1 and IRX5 were significantly elevated.Compared with the model group,the tumor volume of Jianpi Xiaoyou Formula high-dose group,low-dose group and 5-Fu group decreased significantly(P<0.01),and the tumor inhibition effect was more obvious with the increase of Jianpi Xiaoai Formula concentration(P<0.01).The methylation level of m6A in Jianpi Xiaoai Formula high dose group,low dose group and 5-Fu group decreased significantly(P<0.01).The SRAMP website predicted that IRX5 contained multiple m6A sites.Overexpression of METTL3 promoted the expression of IRX5 protein(P<0.001),while knockdown of METTL3 inhibited the expression of IRX5 protein(P<0.001).The drug-containing serum of Jianpi Xiaoai Formula could inhibit the protein expression of METTL3 and IRX5(P<0.05)and inhibit the proliferation,migration and invasion of HCT116(P<0.01).Overexpression of METTL3 and IRX5 reversed the inhibitory effect of Jianpi Xiaoai Formula on HCT116 evil phenotype(P<0.01).Conclusion Jianpi Xiaoai Formula may inhibit METTL3 expression mediated IRX5 low expression to inhibit the progression of colorectal cancer.

Objective By observing the regulatory effect of Strengthening Spleen and Eliminating Cancer Formula on N6-methyladenosine(m6A)methyltransferase,To explore the effect of Strengthening Spleen and Eliminating Cancer Formula on inhibiting the IRX5 m6A level in colorectal cancer(CRC),the regulatory effect on N6-methyladenosine(m6A)methyltransferase was observed.Methods Clinically,m6A hypermethylated genes in colorectal cancer was analyzed by m6A sequencing of pathological tissues from five CRC patients after radical surgery,looking for protein detection indexes for validation.23 BALB/c nude mice were selected and injected with HCT116 cells to establish a nude-mouse transplantation model of human colorectal cancer.They were divided into Model group,Western medicine group(5-fluorouracil group),Chinese medicine group(Jianpi Xiaoai Formula low-dose group,Jianpi Xiaoai Formula high-dose group),with 6 rats in each group,5 rats in control group.The tumor volume of all groups was compared.The overall methylation level of m6A was detected by colorimetric method.The protein expression levels of METTL3,METTL14,and WTAP,in tumor were detected by Western blot.The SRAMP website was used to predict the m6A sites of IRX5.HCT116 cells were treated with oe-NC,oe-METTL3,sh-NC,and sh-METTL3.The expression of IRX5 protein was detected by Western blot.HCT116 cell line was treated with Jianpi Xiaoai Formula drug-containing serum,and transfected with oe-METTL3 and oe-IRX5.The group was set as followed:control group,Jianpi Xiaoai Formula drug-containing serum group,Jianpi Xiaoai Formula drug-containing serum group+oe-NC,Jianpi Xiaoai Formula drug-containing serum group+oe-METTL3,Jianpi Xiaoai Formula drug-containing serum group+oe-IRX5,cell cloning experiment and Transwell experiment were performed to detect cell proliferation,migration and invasion ability of each group.The protein expression levels of METTL3 and IRX5 were detected by Western blot.Results The results of m6A sequencing of genes showed that the m6A methylation level increased in patients with CRC,and the m6A methylation levels of SOX1 and IRX5 were significantly elevated.Compared with the model group,the tumor volume of Jianpi Xiaoyou Formula high-dose group,low-dose group and 5-Fu group decreased significantly(P<0.01),and the tumor inhibition effect was more obvious with the increase of Jianpi Xiaoai Formula concentration(P<0.01).The methylation level of m6A in Jianpi Xiaoai Formula high dose group,low dose group and 5-Fu group decreased significantly(P<0.01).The SRAMP website predicted that IRX5 contained multiple m6A sites.Overexpression of METTL3 promoted the expression of IRX5 protein(P<0.001),while knockdown of METTL3 inhibited the expression of IRX5 protein(P<0.001).The drug-containing serum of Jianpi Xiaoai Formula could inhibit the protein expression of METTL3 and IRX5(P<0.05)and inhibit the proliferation,migration and invasion of HCT116(P<0.01).Overexpression of METTL3 and IRX5 reversed the inhibitory effect of Jianpi Xiaoai Formula on HCT116 evil phenotype(P<0.01).Conclusion Jianpi Xiaoai Formula may inhibit METTL3 expression mediated IRX5 low expression to inhibit the progression of colorectal cancer.

Objective To investigate the needs of the main caregivers of patients with spinal cord injury following neurogenic bowel dysfunction to participate in the whole process of intestinal management.Methods Nine main caregivers of patients with spinal cord injury in Jiangsu Province Hospital from January to Decem-ber,2024 were selected with objective sampling method.They were investigated face-to-face semi-structured in-depth interviews,and the data were analyzed,summarized and extracted by Colaizzi 7-step analysis method.Results Three themes and eight sub-themes were extracted:the needs of professional knowledge and skills standards in intestinal management(lack of intestinal training knowledge,lack of ability to acquire knowledge and different standards of medical institutions);the needs of physical ability and decision making ability participating in intesti-nal training(lack of participation,single decision-making behavior,excessive dependence on medical personnel);the desire for therapeutic benefits and the needs for building psychological confidence(concerns about the pa-tient's prognosis,overdependence on medication).Conclusion Clinical medical staff should pay attention to the actual needs and difficulties faced by the main caregivers of patients with neurogenic bowel dysfunction to participate in the entire intestinal management,and construct targeted training programs,strengthen the skills and knowledge training,to enhance their autonomy and responsi-bility,to achieve the rehabilitation goal better.

Objective To investigate the needs of the main caregivers of patients with spinal cord injury following neurogenic bowel dysfunction to participate in the whole process of intestinal management.Methods Nine main caregivers of patients with spinal cord injury in Jiangsu Province Hospital from January to Decem-ber,2024 were selected with objective sampling method.They were investigated face-to-face semi-structured in-depth interviews,and the data were analyzed,summarized and extracted by Colaizzi 7-step analysis method.Results Three themes and eight sub-themes were extracted:the needs of professional knowledge and skills standards in intestinal management(lack of intestinal training knowledge,lack of ability to acquire knowledge and different standards of medical institutions);the needs of physical ability and decision making ability participating in intesti-nal training(lack of participation,single decision-making behavior,excessive dependence on medical personnel);the desire for therapeutic benefits and the needs for building psychological confidence(concerns about the pa-tient's prognosis,overdependence on medication).Conclusion Clinical medical staff should pay attention to the actual needs and difficulties faced by the main caregivers of patients with neurogenic bowel dysfunction to participate in the entire intestinal management,and construct targeted training programs,strengthen the skills and knowledge training,to enhance their autonomy and responsi-bility,to achieve the rehabilitation goal better.

Objective:To assess the cytotoxicity of self-adhesive resin cement RelyXTM U200 on human periodontal ligament fibro-blasts(HPDLFs)under different light treatment modes.Methods:The self-adhesive resin cement RelyXTM U200 was cured by immediate light(A),intermittent light(B),delayed light(C)and no light(D)treatment respectively.The HPDLFs were cultured with the specimen extracts under different light modes.The proliferation ability and apoptosis level of the cells in the groups were detected by CCK-8 test and flow cytometry apoptosis test.SPSS 20.0 software was used to analyze the cytotoxicity of specimens to HPDLFs.Results:At 24 h after light treatment the cytoxicity(grade)of group A,B,C and D was 1,2,3 and 4,at 72 h 1,2,2 and 3,respectively.At 72 h,the apoptosis index(％)of the cells in A,B,C and D groups was 6.38％±0.94％,16.34％±1.67％,24.13％±1.43％and 38.34％±2.75％respecitvely.Conclusion:The cytotoxicity of self-adhesive resin cement is the greatest under no light treatment mode and the least under immediate light treatment mode.

Objective To investigate the effect of iRoot BP Plus on the repair of dental pulp cavity during pulp injury in rats,and to explore the possible role of iRoot BP Plus in the process of damage repair.Methods Bilateral maxillary first molars of 60 8-week-old SD rats were treated with direct pulp capping after pulp opening.They were divided into iRoot BP Plus group(BP group)and control group.After histological processing,the damage of dental pulp tissue and the calcification of pulp cavities were observed by microCT and HE staining.mRNA was extracted from rat dental pulp tissue,and the expression of inflammation and mineralization genes were detected by real-time quantitatitive PCR.Results The results of this study showed that the dental pulp tissue had different degrees of calcification at 1,4 and 7 days after direct pulp capping.There was no significant difference between the BP group and the control group on the 1st day after the operation,but on the 4th and 7th day after the operation,a regular calcified bridge was formed at the pulp hole in the BP group,and more pulp tissue was retained in the pulp cavity.In the control group,a large area of irregular calcified tissue was formed below the pulp hole.HE staining results showed that the pulp tissue had a heavy inflammatory response,and a large number of inflammatory cells infiltrated near the pulp orifice on the first day after the operation.On the 4th and 7th days after the opera-tion,mineralized tissue began to form near the pulp orifice in the BP group,and healthier pulp tissue was retained in the pulp cavity.The gene levels of IL-1β and TNF-α in the BP group were lower than those in the control group,and the gene expression levels of min-eralization indexes COL1 and DSPP were higher than those in the control group.The difference was statistically significant(P＜0.05).Conclusion Dental pulp injury can induce the immune defense response.If the inflammation is not well controlled,it will stimulate the calcification of dental pulp cavities.iRoot BP Plus can better regulate the dental pulp microenvironment,control the development of inflammation,reduce the formation of adverse calcified masses in the pulp cavity after pulp injury,and preserve more pulp tissue.

Maxillary first molar teeth usually have 4 cusps,a few teeth have stuck cusps on the lingual side of the mesial tongue tip,while the presence of additional cusp on the buccal side of the mesial buccal tip is rarely reported,and it is even more rare for the cusp to connect with the dentin,cementum and pulp cavity of the main crown and fuse together to form C-shaped roots.A case of the buccal supernumerary cusp of the right maxillary first molar with C-shaped root canal was reported.