The postnatal injection of AAV9-FOXG1 rescues corpus callosum agenesis and other brain deficits in the mouse model of FOXG1 syndrome.

Heterozygous mutations in the FOXG1 gene manifest as FOXG1 syndrome, a severe neurodevelopmental disorder characterized by structural brain anomalies, including agenesis of the corpus callosum, hippocampal reduction, and myelination delays. Despite the well-defined genetic basis of FOXG1 syndrome, therapeutic interventions targeting the underlying cause of the disorder are nonexistent. In this study, we explore the therapeutic potential of adeno-associated virus 9 (AAV9)-mediated delivery of the FOXG1 gene. Remarkably, intracerebroventricular injection of AAV9-FOXG1 to Foxg1 heterozygous mouse model at the postnatal stage rescues a wide range of brain pathologies. This includes the amelioration of corpus callosum deficiencies, the restoration of dentate gyrus morphology in the hippocampus, the normalization of oligodendrocyte lineage cell numbers, and the rectification of myelination anomalies. Our findings highlight the efficacy of AAV9-based gene therapy as a viable treatment strategy for FOXG1 syndrome and potentially other neurodevelopmental disorders with similar brain malformations, asserting its therapeutic relevance in postnatal stages.

The patient-specific mouse model with Foxg1 frameshift mutation uncovers the pathophysiology of FOXG1 syndrome.

Single allelic mutations in the gene encoding the forebrain-specific transcription factor FOXG1 lead to FOXG1 syndrome (FS). Patient-specific animal models are needed to understand the etiology of FS, as FS patients show a wide spectrum of symptoms correlated with location and mutation type in the FOXG1 gene. Here we report the first patient-specific FS mouse model, Q84Pfs heterozygous (Q84Pfs-Het) mice, mimicking one of the most predominant single nucleotide variants in FS. Intriguingly, we found that Q84Pfs-Het mice faithfully recapitulate human FS phenotypes at the cellular, brain structural, and behavioral levels. Importantly, Q84Pfs-Het mice exhibited myelination deficits like FS patients. Further, our transcriptome analysis of Q84Pfs-Het cortex revealed a new role for FOXG1 in synapse and oligodendrocyte development. The dysregulated genes in Q84Pfs-Het brains also predicted motor dysfunction and autism-like phenotypes. Correspondingly, Q84Pfs-Het mice showed movement deficits, repetitive behaviors, increased anxiety, and prolonged behavior arrest. Together, our study revealed the crucial postnatal role of FOXG1 in neuronal maturation and myelination and elucidated the essential pathophysiology mechanisms of FS.

Trends of Dental Treatment under General Anesthesia and Patterns of Repeated General Anesthesia in Children and Adolescents with Severe Disabilities / 대한소아치과학회지

The aim of this retrospective study was to evaluate the demographic characteristics of pediatric dental patients who underwent dental treatment under general anesthesia (DTGA) at the Seoul National University Dental Hospital from January 2011 through December 2020 and compare the patterns of repeated DTGA between dental patients with severe disabilities (DSD) and non-DSD (healthy or medically compromised patients without DSD). There were 1,857 DTGAs among 1,719 patients (mean age = 5.1 years; males = 59.3%; ASA 2 or above = 52.9%; DSD = 26.8%). Overall, 6.6% of patients underwent repeated DTGA, and the rate of repeated DTGA over a 10-year period was 7.4%. ASA 2 or above (p < 0.0001) and DSD (p < 0.0001) were more likely to undergo repeated DRGA compared to ASA 1 and non-DSD. At both GA1 and GA2, DSD received significantly more restorative treatment on permanent teeth than non-DSD (p = 0.002, p < 0.0001, respectively). There has been an increasing demand for DTGA in pediatric dentistry over the last 10 years. Regular check-ups and preventive oral health care are necessary for pediatric dental patients with severe disabilities to reduce the possibility of repeated DTGA.

Qualitative Study on the Narcotics Information Management System (NIMS) Experience of Doctors and Pharmacists Using Narcotic Analgesics

Background@#With the increasing use of narcotic analgesics, the Ministry of Food and Drug Safety has been operating the National Narcotics Information Management System (NIMS) since 2018 in Korea. Some studies are related to the narcotic analgesics use, but the evaluation studies of system, including NIMS users are insufficient. @*Objective@#This study evaluated the NIMS enforcement process through in-depth interviews on the experience of using NIMS of doctors and pharmacists who prescribe or dispense opioid analgesics. @*Methods@#Participants in this study were selected through purposeful sampling as three doctors and two pharmacists in the metropolitan area. The interviews were conducted from August 27 to October 19, 2022. Each interview was analyzed using a phenomenological method. @*Results@#Six components of this study included “Doctors and pharmacists come across drug abusers”, “They respond individually to drug abusers”, “They use NIMS tailored to the needs of system managers”, “They are concerned about the role of NIMS in practice settings”, “They face challenges against drug abusers” and “They find a way to utilize the system to deal with drug abusers.” The interviews confirmed that doctors and pharmacists used NIMS little in their work, and they only reported data. Therefore, it might be difficult for doctors and pharmacists to take measures for the safe use of narcotic analgesics. @*Conclusion@#It is likely that adding data-feedback and medication-check function can reduce the abuse of narcotic analgesics. In the future, further studies on the evaluation of the empirical results of NIMS and review of systems in other countries will be required.

Behavioral Phenotypes of Foxg1 Heterozygous Mice.

FOXG1 syndrome (FS, aka a congenital variant of Rett syndrome) is a recently defined rare and devastating neurodevelopmental disorder characterized by various symptoms, including severe intellectual disability, autistic features, involuntary, and continuous jerky movements, feeding problems, sleep disturbances, seizures, irritability, and excessive crying. FS results from mutations in a single allele of the FOXG1 gene, leading to impaired FOXG1 function. Therefore, in establishing mouse models for FS, it is important to test if heterozygous (HET) mutation in the Foxg1 gene, mimicking genotypes of the human FS individuals, also manifests phenotypes similar to their symptoms. We analyzed HET mice with a null mutation allele in a single copy of Foxg1, and found that they show various phenotypes resembling the symptoms of the human FS individuals. These include increased anxiety in the open field as well as impairment in object recognition, motor coordination, and fear learning and contextual and cued fear memory. Our results suggest that Foxg1 HET mice recapitulate at least some symptoms of the human FS individuals.

Bap1/SMN axis in Dpp4+ skeletal muscle mesenchymal cells regulates the neuromuscular system.

The survival of motor neuron (SMN) protein is a major component of the pre-mRNA splicing machinery and is required for RNA metabolism. Although SMN has been considered a fundamental gene for the central nervous system, due to its relationship with neuromuscular diseases, such as spinal muscular atrophy, recent studies have also revealed the requirement of SMN in non-neuronal cells in the peripheral regions. Here, we report that the fibro-adipogenic progenitor subpopulation expressing Dpp4 (Dpp4+ FAPs) is required for the neuromuscular system. Furthermore, we also reveal that BRCA1-associated protein-1 (Bap1) is crucial for the stabilization of SMN in FAPs by preventing its ubiquitination-dependent degradation. Inactivation of Bap1 in FAPs decreased SMN levels and accompanied degeneration of the neuromuscular junction, leading to loss of motor neurons and muscle atrophy. Overexpression of the ubiquitination-resistant SMN variant, SMNK186R, in Bap1-null FAPs completely prevented neuromuscular degeneration. In addition, transplantation of Dpp4+ FAPs, but not Dpp4- FAPs, completely rescued neuromuscular defects. Our data reveal the crucial role of Bap1-mediated SMN stabilization in Dpp4+ FAPs for the neuromuscular system and provide the possibility of cell-based therapeutics to treat neuromuscular diseases.

BAP1 shapes the bone marrow niche for lymphopoiesis by fine-tuning epigenetic profiles in endosteal mesenchymal stromal cells.

Hematopoiesis occurs within a unique bone marrow (BM) microenvironment, which consists of various niche cells, cytokines, growth factors, and extracellular matrix components. These multiple components directly or indirectly regulate the maintenance and differentiation of hematopoietic stem cells (HSCs). Here we report that BAP1 in BM mesenchymal stromal cells (MSCs) is critical for the maintenance of HSCs and B lymphopoiesis. Mice lacking BAP1 in MSCs show aberrant differentiation of hematopoietic stem and progenitor cells, impaired B lymphoid differentiation, and expansion of myeloid lineages. Mechanistically, BAP1 loss in distinct endosteal MSCs, expressing PRX1 but not LEPR, leads to aberrant expression of genes affiliated with BM niche functions. BAP1 deficiency leads to a reduced expression of pro-hematopoietic factors such as Scf caused by increased H2AK119-ub1 and H3K27-me3 levels on the promoter region of these genes. On the other hand, the expression of myelopoiesis stimulating factors including Csf3 was increased by enriched H3K4-me3 and H3K27-ac levels on their promoter, causing myeloid skewing. Notably, loss of BAP1 substantially blocks B lymphopoiesis and skews the differentiation of hematopoietic precursors toward myeloid lineages in vitro, which is reversed by G-CSF neutralization. Thus, our study uncovers a key role for BAP1 expressed in endosteal MSCs in controlling normal hematopoiesis in mice by modulating expression of various niche factors governing lymphopoiesis and myelopoiesis via histone modifications.

Evaluation of the effectiveness of the policy to expand the scope of national health insurance dental scaling service benefits / 대한구강보건학회지

Objectives@#Korea’s National Health Insurance (NHI) coverage rate for dental services is 16.0%, far lower than the 33.0% average of OECD (Organization for Economic Co-operation and development) countries. In 2013, the government implemented a policy to pay for dental scaling service for adults over the age of 20. Then in 2017, the eligibility criteria was expanded to adults aged 19 or older the eligibility age was lowered expanded to 19 or older. @*Methods@#The study aimed to determine the effectiveness of the policy by comparing the number of times per person the dental scaling service was used before and after the implementation of the new applicants group compared to the existing application/applicants group. The analysis was conducted among patients aged 19 and 20 who visited the hospital with gingivitis and periodontal disease and who used the dental scaling service more than once per year as a preventive measure, using the customized database of the NHIC (National Health Insurance Corporation). As of July 1, 2017 when the dental scaling service was implemented by the National Health Insurance Service, the period was set as “before enforcement” (January 2016 to June 2017) and “after enforcement” (July 2017 to December 2018). To evaluate the policy effectiveness of expandingthe ages of individuals eligible for the dental scaling service, Difference-in-Differences (DiD) analysis was conducted to examine the number of times per person the service was used, changes in personal contributions, and policy effectiveness. @*Results@#Since the National Health Insurance policy to pay for dental scaling services, both the applicants group and the sustainable application group have increased the rate of inspection of preventive dental scaling services (19.82%→30.91%, 29.68%→31.18%). As a result of determining the pure effect of the NHI’s policy of covering the dental scaling service, it was found that the the number of times the dental scaling service was used per person increased significantly in the new application group (0.03 times, <.0001). @*Conclusions@#However, it was found that the cost of the dental scaling service per person while the cost of the dental scaling decreased was shown to decrease, it was not statistical significant. Therefore, additional policy support such as lowering the personal financial burden is needed for those who are unable to afford the service.

Health Security Ideas of Major Political Groups and the US Military Government during the Liberation Period (1945-1948) in Korea / 의사학

The liberation period in Korea was when creative imagination and various debates existed about plans for political, economic, and social systems. Among them was the debate over the national health security underlying the social safety net. Although the US influenced the Korean health security after liberation, major political groups on the Korean peninsula also expressed various opinions. However, previous studies have shown little interest in national health security, which operates the public health and medical care systems. To overcome these limitations, this study focuses on the ideas on national health security presented by major political groups, analyzing the reply proposal of “Jŏnpyŏng” and the health care proposal of the US military government, which has not been reviewed before.The opinions of major political groups including the right-wing Im-hyŏp and left-wing Min-chŏn diverged on national health security issue regarding insurance coverage, measures to secure financial resources, items of insurance benefits, and measures to stabilize the supply and demand of medical personnel. The claims of the US military government can be understood by “Labor Problems and Policies in Korea (Korean Subcommittee),” “Korean Labor Report (Stewart Meacham),” and “Proposed Political Platform Provisional Korean Democratic Government (Sub-commission #2).”The major political groups and the US military government agreed on the need for social protection against death, old age, disability, disease, injury, and unemployment. All of them claimed national health security, in which the roles of the private sector and the government were mixed, should be gradually introduced. The major political groups, in particular, proposed to (1) set workers as beneficiaries of insurance, (2) share financial resources jointly among the state, employers, and workers, and (3) promote the expansion of the number of doctors and medical institutions and prefer cooperative operations of the hospitals established in small administrative units.This paper argues that the ideas on national health security during the liberation period did not completely deviate from the global trend immediately after World War II when countries tried to expand the number of people covered by national health security and strengthen its coverage. Although these ideas were not fully reflected in the Constitution of 1948, it is significant in that the Constitution codified for the first time the state’s responsibility for those who have no ability for living due to their health conditions.

How general anesthetics work: from the perspective of reorganized connections within the brain / 대한마취과학회지

General anesthesia is critical for various procedures and surgeries. Despite the widespread use of anesthetics, their precise mechanisms remain poorly understood. Anesthetics inevitably act on the brain, primarily through the modulation of target receptors. Even if the action is specific to an individual neuron, however, long-range effects can occur due to the tremendous interconnectedness of neuronal activity. The strength of this connectivity can be understood using mathematical models that allow for the study of neuronal connectivity dynamics. These models also allow researchers to develop hypotheses on the candidate mechanisms of action of different types of anesthesia. This review highlights the theoretical background associated with the study of the mechanisms of action of anesthetics. We propose a candidate framework that describes how anesthetics act on the brain and consciousness in general.

A Multi-level Analysis of Factors Affecting Participation in Health Screenings in Korea: A Focus on Household and Regional Factors / 예방의학회지

Objectives@#This study divided the factors that affect participation in health screenings into individual, household, and regional levels and conducted a multi-level analysis to identify the factors related to participation in health screenings. @*Methods@#Participants from the 2017 Community Health Survey were classified into 2 groups (under 40 and 40 or older). A multi-level logistic regression analysis was conducted to identify the factors that affected participation in health screenings. @*Results@#The screening rate of the participants was 69.7%, and it was higher among participants aged 40 and older (80.3%) than it was among participants younger than 40 (49.8%). At the individual level, the factors that influenced participation in health screenings included age, economic activity, smoking status, physician-diagnosed hypertension, and a moderate or high physical activity level. At the household level, the odds ratio of participation in health screenings was high for participants who lived in single-person households, lived with a spouse, earned a high monthly household income, and were not beneficiaries of national basic livelihood security. At the regional level, the odds ratio at the 95% confidence interval level of participation in health screenings was high for participants who had trust in the local community and lived in an area with a proportionally high social welfare budget. @*Conclusions@#This study analyzed nationalwide data and confirmed that individual, household, and regional characteristics affected participation in health screenings. Therefore, policies that prioritize the improvement of regional level factors and especially household level factors are likely to be the most effective for improving the screening rate.

Detection of Proximal Caries Lesions with Deep Learning Algorithm / 대한소아치과학회지

This study aimed to evaluate the effectiveness of deep convolutional neural networks (CNNs) for diagnosis of interproximal caries in pediatric intraoral radiographs. A total of 500 intraoral radiographic images of first and second primary molars were used for the study. A CNN model (Resnet 50) was applied for the detection of proximal caries. The diagnostic accuracy, sensitivity, specificity, receiver operating characteristic (ROC) curve, and area under ROC curve (AUC) were calculated on the test dataset. The diagnostic accuracy was 0.84, sensitivity was 0.74, and specificity was 0.94. The trained CNN algorithm achieved AUC of 0.86. The diagnostic CNN model for pediatric intraoral radiographs showed good performance with high accuracy. Deep learning can assist dentists in diagnosis of proximal caries lesions in pediatric intraoral radiographs.

The histone H3-lysine 4-methyltransferase Mll4 regulates the development of growth hormone-releasing hormone-producing neurons in the mouse hypothalamus.

In humans, inactivating mutations in MLL4, which encodes a histone H3-lysine 4-methyltransferase, lead to Kabuki syndrome (KS). While dwarfism is a cardinal feature of KS, the underlying etiology remains unclear. Here we report that Mll4 regulates the development of growth hormone-releasing hormone (GHRH)-producing neurons in the mouse hypothalamus. Our two Mll4 mutant mouse models exhibit dwarfism phenotype and impairment of the developmental programs for GHRH-neurons. Our ChIP-seq analysis reveals that, in the developing mouse hypothalamus, Mll4 interacts with the transcription factor Nrf1 to trigger the expression of GHRH-neuronal genes. Interestingly, the deficiency of Mll4 results in a marked reduction of histone marks of active transcription, while treatment with the histone deacetylase inhibitor AR-42 rescues the histone mark signature and restores GHRH-neuronal production in Mll4 mutant mice. Our results suggest that the developmental dysregulation of Mll4-directed epigenetic control of transcription plays a role in the development of GHRH-neurons and dwarfism phenotype in mice.

Detecting of Proximal Caries in Primary Molars using Pen-type QLF Device / 대한소아치과학회지

The purpose of this in vivo study was to assess the clinical screening performance of a quantitative light-induced fluorescence (QLF) device in detecting proximal caries in primary molars. Fluorescence loss, red autofluorescence and a simplified QLF score for proximal caries (QS-proximal) were evaluated for their validity in detecting proximal caries in primary molars compared to bitewing radiography.Three hundred and forty-four primary molar surfaces were included in the study. Carious lesions were scored according to lesion severity assessed by visual-tactile and radiographic examinations. The QLF images were analyzed for two quantitative parameters, fluorescence loss and red autofluorescence, as well as for QS-proximal. For both quantitative parameters and QS-proximal, the sensitivity, specificity and area under receiver operating curve (AUROC) were calculated as a function of the radiographic scoring index at enamel and dentin caries levels.Both quantitative parameters showed fair AUROC values for detecting dentine level caries (△F = 0.794, △R = 0.750). QS-proximal showed higher AUROC values (0.757 - 0.769) than that of visual-tactile scores (0.653) in detecting dentine level caries.The QLF device showed fair screening performance in detecting proximal caries in primary molars compared to bitewing radiography.

A 20-year Retrospective Study of Pediatric Oral Lesion Biopsy / 대한소아치과학회지

The aim of this study was to provide pathological information of pediatric oral lesions by retrospectively analyzing oral biopsy results from pediatric patients at the Seoul National University Dental Hospital.Biopsy results of all oral lesions from pediatric patients, aged 0 - 16 years, were collected from the files of the Department of Oral Pathology, Seoul National University Dental Hospital from January 2000 to April 2020. The patients were divided into 3 age groups: 0 - 5, 6 - 11 and 12 - 16 years. All oral lesions were classified into three main categories: inflammatory and reactive, tumor or tumor-like and cystic lesions.Among the total of 2928 biopsy specimens, tumor or tumor-like lesions(35.66%) were the most common, followed by inflammatory and reactive lesions(34.29%) and cystic lesions(30.05%). Regardless of the categories used in this study, odontoma was the most frequently found lesion, mucocele and dentigerous cyst being the next common. This study was the first retrospective review of pediatric oral pathology in Korea, and the results from this study may assist in providing informative insight into the pediatric oral pathology for pediatric dentists.

Root and Canal Morphology of Maxillary Primary Molar using CBCT and 3D CT / 대한소아치과학회지

The purpose of this study is to analyze morphological characteristics of maxillary primary molar’s root and root canal. 268 children aged 3 - 7 years (175 boys, 93 girls) who had CBCT (152 children) and 3D CT (116 children) taken in Seoul National University Dental Hospital from January 2006 to April 2020 were included. The number of roots and root canals were analyzed in 1002 teeth without any root resorption or periapical pathologies. Curvature, angulation, length of root and root canal, as well as cross-sectional shapes of the root canal were analyzed in 218 teeth. By using Mimics and 3-Matics software, volume, surface area, and volume ratio of root canal was analyzed in 48 teeth.More than half of maxillary primary molars have 3 roots and 3 root canals. The degree of symmetry of root canal type was about 0.63 (Cohen’s kappa coefficient). The most frequent shape of roots and canals was linear in 1st primary molars and curved in 2nd primary molars. Angulation, length of root and root canals was the largest on palatal roots. Most teeth showed ovoid or round shapes at apex. The largest root canal volume, surface area, volume ratio was found in the palatal roots.

Clinical Factors Affecting the Results of Regenerative Endodontic Procedure / 대한소아치과학회지

This study retrospectively analyzed the effect of clinical factors on the outcomes of REP(regenerative endodontic procedure). Patients who received the REP using triple antibiotic paste due to trauma or fracture of dens evaginatus from February, 2011 to January, 2020 were included in the study. Finally, 57 teeth in 54 patients were selected.Investigated clinical factors were as follows: intentional bleeding, etiology, and root development stage. Treatment outcomes evaluated were as follows: improvement of subjective symptoms, changes in the periapical lesion, and the amounts of root development after REP. To compensate for differences in angulation and position between repeated radiographic examinations, images were aligned by Turboreg plugin. To evaluate the amounts of root development, apical diameter, root area, and root length were measured by ImageJ software.Among the aforementioned factors, intentional bleeding had no significant effect on treatment results. Regarding the etiology, the increase in the root area and the root length was significantly less in trauma cases than in dens evaginatus fracture cases. Considering root development stage, more immature teeth presented more increase in the root area.

Distribution of Pediatric Malocclusion Patients in Seoul National University Dental Hospital / 대한소아치과학회지

A total of 580 patients, who visited and received an orthodontic diagnosis in the Department of Pediatric Dentistry, Seoul National University Dental Hospital from 2017 to 2019, were investigated in this study. The aim of this study was to evaluate skeletal patterns of pediatric orthodontic patients determined with lateral cephalometric analysis and to analyze the relationship between skeletal pattern and probable associated clinical features. Also, the modality of orthodontic treatment for each skeletal classification was investigated to aid in therapeutic decisions.Patients aged 7 year accounted for the largest age group; 54.2% of patients showed a skeletal class I pattern, 22.2% showed a skeletal class II pattern, and 23.6% showed a skeletal class III pattern. Bi-maxillary retrusion for skeletal class I, retruded mandible with normal positioning of the maxilla for skeletal class II, and retrusion of the maxilla with protrusion of the mandible for skeletal class III were the largest subgroups by skeletal pattern. Brachyfacial type accounted for 55.0% of patients, followed by 31.9% of mesofacial type and 13.1% of dolichofacial type. The prevalence of anterior crossbite in the study was 43.3%, higher than that in previous studies.

Dental Characteristics of Microcephalic Osteodysplastic Primordial Dwarfism Type II / 대한소아치과학회지

Microcephalic osteodysplastic primordial dwarfism type II (MOPD II) is an autosomal recessive inherited disorder form of primordial dwarfism, caused by mutations in the pericentrin gene. The purpose of the study was to examine the clinical and radiological features, physicochemical properties and microstructures of the tooth affected with MOPD II.The mandibular 2nd molar was collected from the MOPD II patient. Micro-computerized tomography, scanning electron microscopy, energy dispersive spectrometry and Vickers microhardness analysis were performed on the MOPD II and the normal control.The morphology of the MOPD II tooth appeared to have malformed pulp and root and showed a small size. The mineral density measurement showed that the MOPD II tooth had similar scores in the enamel, but lower scores in the root 1/2 and apical dentin compared to the normal control. The microhardness values were smaller in the cusp enamel, root 1/2 dentin and apical dentin of the MOPD II compared to the normal control.In this study, the dental characteristics and the physicochemical properties of a tooth affected with MOPD II were analyzed to improve understanding of the oral manifestations of the disease and to assist in proper dental treatment by identifying precautions.

Retrospective study on three-dimensional characteristics of mesiodens using CBCT in pediatric dentistry / 대한소아치과학회지

The purpose of this study was to investigate the three-dimensional characteristics of mesiodens using Cone-beam Computed Tomography(CBCT) and analyze the factors affecting complications and anesthetic methods of extraction. This study evaluated 602 mesiodens of 452 patients who underwent extraction of mesiodens at the department of Pediatric Dentistry in Seoul National University Dental Hospital between 2017 and 2019.The ratio of mesiodens patients over total patient per year was gradually increased over the past 20 years. Mesiodens with labio-palatally horizontal direction while root directing labial were the most common among the mesiodens with horizontal direction. Mesiodens were the most common at the cervical side of the adjacent teeth(37.0%) and mesiodens located in the near-palatal side were observed about 3.83 times higher than the far-palatal side. Most of the mesiodens(82.1%) were in contact with adjacent permanent teeth on all three sides of the CBCT and 46.2% of mesiodens had curved roots. The patient’s age, vertical position, presence of complications, and proximity showed a significant difference in the selection of general anesthesia among anesthetic methods. The direction and vertical position of mesiodens had a significant effect on complications.These results provide a better understanding of mesiodens for establishing an accurate diagnosis and treatment plan.