Objective:Induced pluripotent stem cells (iPSCs) cell lines were established using peripheral blood mononuclear cells (PBMCs) from a patient suffering from neonatal respiratory distress syndrome (NRDS) who carried Adenosine triphosphate-binding cassette transporter A3 ( ABCA3) compound heterozygous mutations. Methods:Cell experimental research.Peripheral venous blood was collected and PBMCs were isolated and cultured in vitro. PBMCs were transfected with non-integrated Sendai vector carrying reprogramming factors.The chromosome karyotypes of the established iPSCs were analyzed.Immunofluorescence and flow cytometry were used to detect pluripotency markers of stem cells and verify their differentiation potential.Sanger sequencing was performed to analyze gene mutations.In addition, short tandem repeat (STR) analysis was performed, polymerase chain reaction(PCR) and agarose gel electrophoresis were used to detect virus residual. Results:Karyotype analysis of established iPSCs cell lines showed normal diploid 46, XY karyotype.Immunofluorescence showed positive staining of stem cell pluripotency markers OCT4, SSEA4, Nanog and Sox2.Flow cytometry was used to detected stem cell pluripotency markers and showed expression of TRA-1-60, SSEA-4 and OCT4.After differentiation into all three germ layers, immunofluorescence was performed to detect ectoderm (Pax-6), mesoderm (Brachyury) and endoderm alpha-fetoprotein markers, and the results showed positive staining, which confirmed that the iPSCs had the potential to differentiate.Sanger sequencing showed c. 3997_3998del and c. 3137C>T compound heterozygous mutations.STR analysis showed they originate from PBMCs, and no Sendai virus residual was detected by PCR and agarose gel electrophoresis.Conclusions:In this study, PBMCs from patient carrying ABCA3 compound heterozygous mutations was used to establish iPSCs cell lines.The research lays a foundation for the study of pathogenesis, therapeutic drug screening and cell therapy of NRDS caused by ABCA3 gene mutations.

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 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.

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 explore the clinical and genetic features of a child with autosomal dominant mental retardation type 40 (MRD40) due to variant of the CHAMP1 gene. METHODS: Clinical characteristics of the child were analyzed. Genetic testing was carried out by low-depth high-throughput and whole genome copy number variant sequencing (CNV-seq) and whole exome sequencing (WES). A literature review was also carried out for the clinical phenotype and genetic characteristics of patients with MRD40 due to CHAMP1 gene variants. RESULTS: The child, a 11-month-old girl, has presented with intellectual and motor developmental delay. CNV-seq revealed no definite pathogenic variants. WES has detected the presence of a heterozygous c.1908C>G (p.Y636*) variant in the CHAMP1 gene, which was carried by neither parent and predicted to be pathogenic. Literature review has identified 33 additional children from 12 previous reports. All children had presented with developmental delay and mental retardation, and most had dystonia (94.1%), delayed speech and/or walking (85.2%, 82.4%) and ocular abnormalities (79.4%). In total 26 variants of the CHAMP1 gene were detected, with all nonsense variants being of loss-of-function type, located in exon 3, and de novo in origin. CONCLUSION The heterozygous c.1908C>G (p.Y636*) variant of the CHAMP1 gene probably underlay the WRD40 in this child. Genetic testing should be considered for children featuring global developmental delay, mental retardation, hypertonia and facial dysmorphism.
Humans ; Intellectual Disability/genetics* ; Genetic Testing ; Phenotype ; Exome Sequencing ; Heterozygote ; Mutation ; Chromosomal Proteins, Non-Histone/genetics* ; Phosphoproteins/genetics*

Objective To analyze the incidence of occupational diseases in Dandong from 2015 to 2019, discuss the characteristics and rules of occupational diseases, and provide theoretical basis for the formulation of occupational disease prevention measures. Methods The occupational disease report data of Dandong from 2015 to 2019 were collected and analyzed by using Excel and SPSS statistical software. Results From 2015 to 2019, a total of 112 new cases of occupational diseases were reported in Dandong, including 108 cases of occupational pneumoconiosis, accounting for 96.43%. Four patients with other occupational diseases, accounting for 3.57% . Among the newly reported cases of occupational diseases, 111 cases were male, accounting for 99.10%, and one case was female, accounting for 0.89%. There were statistically significant differences in the reported conditions of different types of occupational diseases according to gender (P < 0.05). The age of onset of occupational diseases mainly concentrated in 40-69 years old, and there was no statistical significance in the age of onset of different types of occupational diseases (P > 0.05). The length of contact at the onset of the disease was mainly concentrated in 11-20 years, and there was no statistical significance in the comparison of the length of contact at the onset of different types of occupational diseases (P > 0.05). In terms of industry distribution, the mining industry reported the most cases (87 cases, accounting for 77.68%). The private enterprises were more concentrated, accounting for 68 cases, 60.71%. Small and medium-sized enterprises reported the most new cases of occupational diseases, with 107 cases, accounting for 95.54%. There was statistical significance in the new cases of occupational diseases with different industry distribution (P < 0.05). The analysis showed that there was no statistical significance in the new cases of occupational pneumoconiosis in different types of work with the different distribution of length of exposure (P > 0.05). Conclusions Occupational pneumoconiosis accounted for the majority of new cases of occupational diseases in Dandong city from 2015 to 2019, indicating that occupational pneumoconiosis is still the focus of occupational disease prevention and control in Dandong City. In the future occupational disease prevention work, we should actively standardize the implementation of the ^“Law of the People's Republic of China on the Prevention and Control of Occupational Diseases^”, do a good job in the supervision and management of key groups and positions, and effectively protect the health of workers.

Objective:To analyze independent influencing factors of surgical textbook outcome (TO) in patients with gallbladder carcinoma, and to establish a nomogram for predicting TO and evaluated the predictive ability.Methods:Patients with gallbladder carcinoma who underwent surgery in Department of Hepatobiliary and Pancreatic Surgery at Dongfang Hospital Affiliated to Shanghai Tongji University and Department of Biliary Tract Surgery Ⅰ, Third Affiliated Hospital of Naval Medical University (Shanghai Eastern Hepatobiliary Surgery Hospital) from January 2013 to December 2018 were included and the clinical features were retrospectively analyzed. A total of 232 patients were included, including 114 males and 118 females, aged (61.0±9.8) years. According to whether TO reached or not, they were divided into TO group ( n=86) and non-TO group ( n=146). Univariate and multivariate logistic regression were used to analyze the independent influencing factors of TO. The predictive nomogram model of TO was constructed. The receiver operating characteristic (ROC) curve was drawn to evaluate the predictive ability of the model, and the consistency of the predictive model was evaluated by the consistency curve graph and the Hosmer-Lemeshow test. Results:The 1-year and 3-years cumulative survival rates of patients with gallbladder carcinoma in the TO group (86.0% and 62.8%) were better than those in the non-TO group (46.6% and 27.3%), and the difference was statistically significant (χ 2=60.74, P<0.001). In multivariate analysis, higher T stage ( OR=0.16, 95% CI: 0.03-0.79, P<0.001) and cervical gallbladder cancer ( OR=0.14, 95% CI: 0.02-0.94, P=0.004) had the greatest negative association with a TO, and the higher the degree of tumor differentiation ( OR=7.08, 95% CI: 1.34-37.56, P=0.001), the easier it is to achieve TO. The ROC curve showed that the area under the curve of the predictive model was 0.84 (95% CI: 0.79-0.90), suggesting that the model had good predictive performance. A nomogram to assess the probability of TO was developed and had good accuracy in both the consistency curve and Hosmer-Lemeshow test (χ 2=5.77, P=0.673). Conclusion:Tumor T stage, tumor differentiation degree and tumor location are independent influencing factors for achieving TO in patients with gallbladder carcinoma after surgery. The nomogram model constructed according to the above conclusions could accurately predict the probability of reaching TO.

Gastrointestinal cancers are the common malignant tumors of the digestive system, and their morbidity and mortality are in the forefront of malignant tumors. Currently, cancer immunotherapy is the hottest topic in cancer research field. Although cancer immunotherapy has achieved some results in the fundamental research and clinical application of gastrointestinal tumors, there are still a series of problems that need to be resolved. In this article, we review the fundamental and clinical research progress of several common methods of cancer immunotherapy in the field of gastrointestinal tumors.

Background: The incidence of ulcerative colitis (UC) has gradually increased in China in recent years. The pathogenesis of UC is related to the dysfunction of immune system. B7-H5 is an important immune checkpoint molecule and is significant for the regulation of immune function. Ainis: To investigate the expression and clinical significance of B7-H5 in UC. Methods: A total of 65 UC tissue specimens were collected from Jan. 2010 to Dec. 2020 at the First Affiliated Hospital of Soochow University, and 5 healthy subjects were served as controls. Immunohistoehemistry and immunofluorescence were used to detect the expression of B7-H5, and its relationship with elinieopathologieal characteristics of UC patients was analyzed. Results: Expression of B7-H5 was significantly increased in UC patients than in controls (P < 0. 001). B7-H5 expression in UC patients was positively correlated with ESR and CRP (P < 0. 01), but not related to gender, age, extent of lesion, Mayo score and UCEIS score (P > 0. 05). Conclusions; The expression of B7-H5 in UC patients is significantly increased and is correlated with ESR and CRP, and can be used as a new marker for reflecting the severity of inflammation in UC patients.