ObjectiveTo investigate the intervention effect of Jiedu Tongluo Tiaogan prescription （JTTP） in protecting pancreatic β cells by targeting the bile acid Takeda G protein-coupled receptor 5 （TGR5）/cyclic adenosine monophosphate （cAMP） signaling pathway against NOD-like receptor protein 3 （NLRP3） inflammasome. MethodThirty-two male SPF-grade db/db mice were randomly divided into the model group， low-dose JTTP group （3.6 g·kg^-1）， high-dose JTTP group （7.2 g·kg^-1）， and metformin group （0.2 g·kg^-1）. Eight db/m mice were assigned to the blank control group. The mice were treated with drugs for 8 weeks， and fasting blood glucose （FBG） was measured every 2 weeks. Oral glucose tolerance tests （OGTT） were conducted after the last administration. Enzyme-linked immunosorbent assay （ELISA） was performed to detect fasting insulin （FINS）， and the homeostasis model assessment of β-cell function （HOMA-β）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， and IL-1β levels were calculated. Hematoxylin-eosin （HE） staining was used to observe pathological changes in mouse pancreatic tissue. Immunofluorescence was performed to detect insulin expression in mouse pancreatic tissue. Western blot and real-time quantitative polymerase chain reaction （Real-time PCR） were used to detect the expression of proteins and mRNAs of key targets in the TGR5/cAMP signaling pathway and NLRP3 inflammasome. ResultCompared with blank group， FBG， OGTT， FINS， IL-6， TNF-α and IL-1β in model group were significantly increased （P<0.01）. Compared with model group， after 6 weeks of drug treatment， FBG level in JTTP group and metformin group decreased significantly （P<0.01）. The results of OGTT experiment showed that compared with model group， the blood glucose levels of mice in each administration group were decreased at all time points （P<0.05， P<0.01）， and the levels of FINS， TNF-α and IL-6 in JTTP dose groups and metformin group were significantly decreased. The level of IL-1β in JTTP high-dose group and metformin group was significantly decreased （P<0.01）. Pancreatic pathology showed that the islets in the model group were irregular in shape， uneven in distribution， and showed signs of atrophy. The prognosis of JTTP was that the cell count increased and the boundary was clearer. Immunofluorescence results showed that the islet cells in the blank group were arranged in an orderly and full shape with appropriate insulin secretion， while the islet cells in model group were distorted in shape， atrophy in structure and less insulin secretion. The insulin content of mice in JTTP and metformin group was significantly increased. Compared with blank group， mRNA expressions of NLRP3， apoptosis-related spot-like protein （ASC） and Caspase-1 in pancreatic tissues of model group were significantly increased （P<0.01）. Compared with model group， JTTP high-dose group and metformin group promoted the up-regulation of TGR5 and cAMP mRNA， and down-regulated the mRNA expressions of NLRP3， ASC and Caspase-1 （P<0.05， P<0.01）. Compared with blank group， the expression of TGR5 protein in model group was significantly decreased （P<0.01）. Compared with model group， TGR5 protein in JTTP high-dose group and metformin group was significantly increased （P<0.01）.

Objective To explore the automated identification and diameter measurement methods for inferior vena cava (IVC) based on clinical ultrasound images of IVC. Methods An automated identification and localization method based on topology and automatic tracking algorithm was proposed. Tracking algorithm was used for identifying and continuously locating to improve the efficiency and accuracy of measurement. Tests were conducted on 18 sets of ultrasound data collected from 18 patients in intensive care unit (ICU),with clinicians' measurements as the gold standard. Results The recognition accuracy of the automated method was 94.44％ (17/18),and the measurement error of IVC diameter was within the range of±1.96s (s was the standard deviation). The automated method could replace the manual method. Conclusion The proposed IVC automated identification and localization algorithm based on topology and automatic tracking algorithm has high recognition success rate and IVC diameter measurement accuracy. It can assist clinicians in identifying and locating IVC,so as to improve the accuracy of IVC measurement.

Dong qi,referring to beating beside the navel,was proposed for the first time by Nan Jing."Beating above the navel""beating below the navel",and"beating below the heart"in Shanghan Lun all belong to the category of Dong qi.The pathogenesis of Dong qi can be learned from Shanghan Lun.Yang deficiency of the heart,spleen,and kidney,with qi failing to transform body fluids,results in fluid retention.Fluid retention is the key etiological factor of Dong qi.Although there is an exterior pattern with Dong qi,the exterior-releasing and purgative method cannot be used for treatment,or the patient will be impaired of yang and exhausted of yin,fluid circulation will be disturbed,and so on.Reinforcing yang qi to drain water retention is the treatment principle of Dong qi.The coupled medicines of Fuling and Guizhi were commonly used by ZHANG Zhongjing to treat Dong qi.This reflects the treatment principle of strengthening the spleen and warming yang for transforming qi to circulate body fluids.ZHANG Bi,a physician from the Yuan Dynasty who learned from ZHANG Zhongjing,thought that the treatment of Dong qi was based on the method of cultivating earth to restrain water and taking into account the ascending,descending,entering,and exiting of qi of the liver,heart,lung,and kidney while tonifying the spleen,and then created Baoming Siqi Powder(Zhiqiao,Fuling,Fangkui,and Qianhu Powder)for the treatment of Dong qi,which is of great significance to the clinic.

Objective:To determine the optimal irradiation energy and frequency for the establishment of melasma mouse model using simple ultraviolet irradiation, and to provide guidance on animal strains and irradiation protocols for the successful establishment of melasma model.Methods:Animal models of melasma were established using BALB/c female mice and C57BL/6JNifdc female mice. BALB/c female mice were divided into 4 groups using a simple randomization method: A, B, C and G, with 5 mice in each group. C57BL/6JNifdc female mice were divided into 4 groups: D, E, F and H, with 5 mice in each group. All mice were irradiated with 8.428 mW/cm 2 of ultraviolet light. The irradiation time was 15 s (single irradiation energy of 0.13 J/cm 2) in groups A and D, 15 min (single irradiation energy of 7.59 J/cm 2) in groups B and E, and 30 min (single irradiation energy of 15.17 J/cm 2) in groups C and F. Each cycle consisted of 5 consecutive days of irradiation followed by 2 days of cessation, totaling 4 cycles of irradiation. Groups G and H were not irradiated. At the end of irradiation, all mice were kept under normal conditions. One week later, 3 mice from each group were selected for HE, Masson-Fontana, Masson, and immunohistochemical staining. Quantitative analysis was performed to measure the thickness of the acanthocyte layer, melanin granules, collagen percentage, and interleukin-1 (IL-1) levels. The remaining mice were kept for an additional week, depilated and photographed to observe the changes in coloration. Data were analyzed using SPSS 27.0 software, measurement data that did not conform to normal distribution were represented by M( Q1, Q3) and comparisons between groups were made using the Kruskal-Wallis rank sum test. Results:During the entire irradiation process, no visible discoloration was observed in the BALB/c female mice in all groups. In contrast, varying sizes of discoloration appeared in the C57BL/6JNifdc female mice in groups D, E, and F after irradiation in the second week. However, by the third week, the discoloration in group D gradually disappeared, while the discoloration in group E was more obvious than before. At the same time, group F exhibited significant discoloration, with some mice exhibited signs of skin peeling, burning and breakage on their backs. After the 4th week of irradiation, no new discoloration was formed in group D. The discoloration was more obvious in group E, and most mice in group F showed skin burn breakage. Two weeks after the completion of irradiation, there was no obvious discoloration on the dorsal skin of BALB/c female mice in all groups. In C57BL/6JNifdc female mice, group D showed no obvious discoloration, group E exhibited lighter discoloration compared to the 4th week post-irradiation, and group F had crusted skin at the burn sites with lighter discoloration than before. However, the discoloration in groups E and F was still obviously visible to the naked eye. HE staining showed that the difference in the thickness of the echinocyte layer was not statistically significant in groups A, B, C, and G ( H=1.08, P=0.782); whereas the difference was statistically significant in groups D, E, F and H ( H=12.85, P=0.005). The thickness of the echinocyte layer decreased gradually with the extension of the irradiation time. Additionally, there was a disruption in the arrangement of epidermal spindles in group F, and this situation was not observed in groups D and E. Masson-Fontana staining revealed no significant pigmentation in any of the BALB/c female mice. The difference in melanin granule counts between groups A, B, C, and G was not statistically significant ( H=7.77, P=0.051). In contrast, C57BL/6JNifdc female mice exhibited more noticeable pigmentation in the epidermis and dermis in groups E and F. The difference in melanin particle counts among groups D, E, F and H was statistically significant ( H=17.61, P<0.001), with melanin deposition increasing gradually with the duration of irradiation. Masson staining showed that the difference in collagen percentage between groups A, B, C, and G was not statistically significant ( H=7.26, P=0.064). However, significant disorganization of fibers and a loose structure were observed in groups E and F. The difference in collagen percentage between groups D, E, F, and H was statistically significant ( H=8.65, P=0.034). Immunohistochemical results showed that the difference in IL-1 expression levels between groups A, B, C, and G was statistically significant ( H=17.86, P<0.001); also between groups D, E, F, and H was statistically significant ( H=14.19, P=0.003), suggesting that ultraviolet irradiation stimulated an inflammatory response in the skin of mice. Conclusion:BALB/c female mice are not suitable for melasma models under the frequency and duration of irradiation in this experiment. C57BL/6JNifdc female mice are irradiated with a single irradiation energy dose of 7.59 J/cm 2 five days a week for 4 weeks, which can establish stable animal models of melasma with a specific level of pigmentation that persisted for at least 2 weeks.

Objective:To determine the optimal irradiation energy and frequency for the establishment of melasma mouse model using simple ultraviolet irradiation, and to provide guidance on animal strains and irradiation protocols for the successful establishment of melasma model.Methods:Animal models of melasma were established using BALB/c female mice and C57BL/6JNifdc female mice. BALB/c female mice were divided into 4 groups using a simple randomization method: A, B, C and G, with 5 mice in each group. C57BL/6JNifdc female mice were divided into 4 groups: D, E, F and H, with 5 mice in each group. All mice were irradiated with 8.428 mW/cm 2 of ultraviolet light. The irradiation time was 15 s (single irradiation energy of 0.13 J/cm 2) in groups A and D, 15 min (single irradiation energy of 7.59 J/cm 2) in groups B and E, and 30 min (single irradiation energy of 15.17 J/cm 2) in groups C and F. Each cycle consisted of 5 consecutive days of irradiation followed by 2 days of cessation, totaling 4 cycles of irradiation. Groups G and H were not irradiated. At the end of irradiation, all mice were kept under normal conditions. One week later, 3 mice from each group were selected for HE, Masson-Fontana, Masson, and immunohistochemical staining. Quantitative analysis was performed to measure the thickness of the acanthocyte layer, melanin granules, collagen percentage, and interleukin-1 (IL-1) levels. The remaining mice were kept for an additional week, depilated and photographed to observe the changes in coloration. Data were analyzed using SPSS 27.0 software, measurement data that did not conform to normal distribution were represented by M( Q1, Q3) and comparisons between groups were made using the Kruskal-Wallis rank sum test. Results:During the entire irradiation process, no visible discoloration was observed in the BALB/c female mice in all groups. In contrast, varying sizes of discoloration appeared in the C57BL/6JNifdc female mice in groups D, E, and F after irradiation in the second week. However, by the third week, the discoloration in group D gradually disappeared, while the discoloration in group E was more obvious than before. At the same time, group F exhibited significant discoloration, with some mice exhibited signs of skin peeling, burning and breakage on their backs. After the 4th week of irradiation, no new discoloration was formed in group D. The discoloration was more obvious in group E, and most mice in group F showed skin burn breakage. Two weeks after the completion of irradiation, there was no obvious discoloration on the dorsal skin of BALB/c female mice in all groups. In C57BL/6JNifdc female mice, group D showed no obvious discoloration, group E exhibited lighter discoloration compared to the 4th week post-irradiation, and group F had crusted skin at the burn sites with lighter discoloration than before. However, the discoloration in groups E and F was still obviously visible to the naked eye. HE staining showed that the difference in the thickness of the echinocyte layer was not statistically significant in groups A, B, C, and G ( H=1.08, P=0.782); whereas the difference was statistically significant in groups D, E, F and H ( H=12.85, P=0.005). The thickness of the echinocyte layer decreased gradually with the extension of the irradiation time. Additionally, there was a disruption in the arrangement of epidermal spindles in group F, and this situation was not observed in groups D and E. Masson-Fontana staining revealed no significant pigmentation in any of the BALB/c female mice. The difference in melanin granule counts between groups A, B, C, and G was not statistically significant ( H=7.77, P=0.051). In contrast, C57BL/6JNifdc female mice exhibited more noticeable pigmentation in the epidermis and dermis in groups E and F. The difference in melanin particle counts among groups D, E, F and H was statistically significant ( H=17.61, P<0.001), with melanin deposition increasing gradually with the duration of irradiation. Masson staining showed that the difference in collagen percentage between groups A, B, C, and G was not statistically significant ( H=7.26, P=0.064). However, significant disorganization of fibers and a loose structure were observed in groups E and F. The difference in collagen percentage between groups D, E, F, and H was statistically significant ( H=8.65, P=0.034). Immunohistochemical results showed that the difference in IL-1 expression levels between groups A, B, C, and G was statistically significant ( H=17.86, P<0.001); also between groups D, E, F, and H was statistically significant ( H=14.19, P=0.003), suggesting that ultraviolet irradiation stimulated an inflammatory response in the skin of mice. Conclusion:BALB/c female mice are not suitable for melasma models under the frequency and duration of irradiation in this experiment. C57BL/6JNifdc female mice are irradiated with a single irradiation energy dose of 7.59 J/cm 2 five days a week for 4 weeks, which can establish stable animal models of melasma with a specific level of pigmentation that persisted for at least 2 weeks.

Objective To analyze the stress distributions of two root canal preparation shapes of oval root canals with micro-crack.Methods Twenty single-canal mandibular premolars with oval canals were expanded to create micro-cracks.Roots were sectioned after staining.The generation and distribution of dentin micro-cracks were observed under microscope.Then a finite element(FE)model of sectioned enlarged oval canal roots with micro-cracks was established.The stress distribution of micro-crack and root were analyzed under lateral loading.Results Cracks always appeared in the buccolingual sides of oval canal roots and extended from the intracanal wall to the root surface.This was consistent with the stress concentration on the buccolingual side of the root canal wall shown by FE analysis.When micro-cracks occurred,stresses were transferred to the crack tip and the peak values increased sharply nearly 5 times.This made the cracks propagate easily along this direction,especially in the long axis direction of the tooth.Conclusions The presence of micro-cracks does not change the general stress concentration on root with two preparation morphologies of oval canals.However,the micro-crack causes an extreme stress concentration in the crack tip.This may be the mechanism of rapid propagation of microcracks into vertical root fracture,and dentists need to pay high attention.

Objective To establish a HS-GC test method for the determination of enflurane,isoflurane,diflurane,sevoflurane and its metabolite hexafluoroisopropanol(HFIP)in blood,and to establish a LC-MS/MS method for the determination of trifluoroacetic acid(TFA),the co-metabolite of enflurane,isoflurane,and deflurane in blood.Methods Place 0.5 mL blood sample in a 10 mL headspace bottle,add 1.0 mL ultrapure water to mix,then add 0.5 mL n-butanol internal standard solution,sealed and heated at 70℃for 20 min,take the upper layer of gas for HS-GC analysis,qualitatived by dual-column retention time and quantified by the internal standard curve;Blood sample was acetonitrile precipitated protein,separated by liquid chromatography,scanned with electrospray ionization(ESI),negative ion mode,and examined in multiple reaction monitoring mode(MRM),qualitatived by retention time and characteristic ions,quantified by standard curve.Results The detection limits(LOD)of enflurane,isoflurane,desflurane,heptaflurane,HFIP and TFA are 0.05,0.2,0.5,0.05,0.5 μg/mL and 0.5 ng/mL,and linear range as 1～100 μg/mL(TFA:1～100 ng/mL),with correlation coefficients greater than 0.999.The extraction recovery rate is between 30%and 80%,and the intra-day and inter-day precision are less than 5%.The accuracy is between 85%and 115%.Conclusion This method is quick and simple,and can be used for qualitative and quantitative analysis of enflurane,isoflurane,desflurane,sevoflurane and their metabolites in blood.

Objective:To explore the perceived stress and coping experience of spouses of elderly people with long-term disability and empty nest.Methods:Using phenomenological research methods, semi-structured interviews were conducted with 19 spouses of disabled elderly people in a community in Beijing from February to June 2023, and data analysis was conducted using Colaizzi's 7-step analysis method.Results:The perceived stress and coping experience of spouses of long-term disabled empty nest elderly people were summarized into three themes and ten sub-themes, namely, the dynamic and variable stress perception of spouses (stress changed synchronously with the state of disabled elderly people, stress changed in a "U" shape, and pressure increased in a progressive manner), the challenges for spouses in long-term care of disabled elderly people (difficult in unifying opinions and views, incomprehensible inner emotions, difficulty in ensuring their elderly care, and coexistence of multiple roles and identities) and positive coping experiences and strategies of spouses (actively seeking external support, respecting the subjective wishes of disabled elderly people, and improving dyadic coping abilities) .Conclusions:Community nursing staff should support and guide spouses who undertake care work based on changes in perceived stress characteristics of long-term disabled empty nest elderly people. At the same time, it is necessary to understand the various challenges they face in long-term care, draw on positive coping experiences of spouses, so as to provide a basis for building a friendly family support system for home-based elderly care.

Objective:To develop a virtual reality (VR) exercise rehabilitation system for home-based patients with spinal cord injury, and test patients' acceptance and experience.Methods:The VR exercise rehabilitation system for home-based patients with spinal cord injury was developed by a multidisciplinary team based on the home rehabilitation needs, evidence, and clinical experience of spinal cord injury patients, and further improved after recommendations from experts and patients. From February to July 2024, convenience sampling was used to select 148 patients with spinal cord injuries admitted to the Department of Spinal Cord and Neurological Function Reconstruction at the China Rehabilitation Research Center. Acceptance Questionnaire was used to investigate patients' acceptance of the system. Thirteen patients with spinal cord injuries were selected for semi-structured interviews to explore the themes of their experiences using VR exercise rehabilitation system for home-based patients with spinal cord injury.Results:A VR exercise rehabilitation system for home-based patients with spinal cord injury was developed, which includes four aspects of personal profile, personal assessment, training selection, and care knowledge. The total acceptance score of this system among 148 patients was (87.69±1.59). Through semi-structured interviews, three themes were identified involving excellent experience, expectation of rehabilitation effects, satisfaction with functionality and hope for continuous system updates.Conclusions:The design of VR exercise rehabilitation system for home-based patients with spinal cord injury is scientifically reasonable, which can further verify the rehabilitation effect of the system.

Objective:To detect and analyze the gene variation types of 64 unrelated pedigrees affected with autosomal dominant polycystic kidney disease (ADPKD), and explore the detection efficiency of multiple gene analysis techniques and variation characteristics.Methods:It was a cross-sectional study. The clinical data of 64 pedigrees with ADPKD from Nephrology Department or Genetic and Prenatal Diagnosis Center of the First Affiliated Hospital of Zhengzhou University from December 2017 to August 2020 were retrospectively analyzed. The blood samples of probands and other family members were collected. Genetic analysis was carried out by next generation sequencing, and suspected mutations were verified by multiplex ligation-dependent probe amplification, or long-range PCR combined with Sanger sequencing. Prenatal diagnosis for high-risk fetuses was performed by fetal villi or amniotic fluid cells after genotyping without maternal genomic DNA contamination.Results:Among detected 64 pedigrees, 57 pedigrees (89.06%) had genetic variants in PKD1/PKD2. A total of 49 pathogenic/likely pathogenic variants in PKD1/PKD2 were identified in 51 pedigrees (79.69%), including 14 nonsense variants (28.57%), 14 frameshift variants (28.57%), 11 missense variants (22.45%), 5 splicing variants (10.20%) and 5 deletion variants (10.20%). Of these variants, 87.76% (43/49) were in PKD1 and 12.24% (6/49) were in PKD2. Totally, 14 novel variants in PKD1/ PKD2 were identified, including 7 frameshift variants, 3 splicing variants, 2 nonsense variants, 1 deletion variant and 1 missense variant, of which 11 variants were in PKD1 and 3 variants were in PKD2. Twenty high-risk fetuses from 17 pedigrees received prenatal diagnosis, in whom 6 fetuses had PKD1 variation, and other 14 fetuses had no PKD1/ PKD2-genetic variation. Conclusions:The combination of next-generation sequencing, multiplex ligation-dependent probe amplification, and long-range PCR combined with Sanger sequencing can be helpful for rapid, efficient and accurate genetic diagnosis of ADPKD pedigrees. Point mutations are the most common types in PKD1/PKD2. Fourteen novel variants in PKD1/PKD2 extend its pathogenic variant spectrum and can provide basis for genetic counseling and prenatal diagnosis of ADPKD pedigrees.