[Clinical and genetic characteristics of 21 children with Rubinstein-Taybi syndrome].

Objective: To investigate the phenotypes of Rubinstein-Taybi syndrome (RSTS) caused by variants in the CREBBP or EP300 gene, and the correlation between genotype and phenotype. Methods: This case series study was performed on pediatric patients who were referred to the Children's Hospital of Capital Institute of Pediatrics between January 2013 and July 2022. Both point variant and copy number deletion in CREBBP or EP300 gene were detected by whole exome sequencing, chromosomal microarray analysis, or copy number variation sequencing (CNV-seq). The variant categories were summarized and phenotype numbers were re-visited for RSTS patients. Based on variant types, the patients were divided into different groups (point variant or copy number deletion, EP300 or CREBBP point variant, and loss of function or missense variant). Phenotype counts between different groups were compared using the rank-sum test of two independent samples. Results: A total of 21 RSTS patients were recruited, including 12 males and 9 females, with ages ranging from 1 month to 14 years and 2 months. Among them, 67% (14/21) had point variants, and 33% (7/21) had copy number deletions. Out of these, 20 variants (95%) were de novo. Among 20 patients finishing phenotype count during re-visit, 95% (19/20) of the patients exhibited developmental delays before the age of 2 years. Additionally, 80% (16/20) of the patients had distinctive facial features. Considering phenotype count, no statistically significant difference was found between point variant (14 cases) and copy number deletion (6 cases) (5.0 (3.0, 7.0) vs. 5.0 (2.5, 5.3), Z=0.75, P=0.452), CREBBP (10 cases) and EP300 gene (4 cases) point variant (5.0 (3.8, 7.0) vs. 4.0 (2.0, 6.0), Z=1.14, P=0.253), and loss of function (9 cases) and missense (5 cases) variant (6.0 (4.5, 7.0) vs. 3.0 (2.5, 5.5), Z=1.54, P=0.121). Conclusions: Patients with RSTS primarily exhibit developmental delays in early childhood. Specific facial features serve as suggested signs of genetic testing. However, no significant genotype-phenotype correlation is found.

[Review on the development of periodontology in China].

Chinese Journal of Stomatology has gone through 70 years of ups and downs, witnessing the development of periodontics in China from a faltering start to twists and turns, and finally innovative development. This article aims to review the periodontology-related papers published in the Chinese Journal of Stomatology. Based on the characteristics of the times, they are summarized into five stages: staggering start, forced stagnation, vigorous development, standardized innovation, and disciplinary integration. Researches on periodontal diseases in China initially focused on learning and reference, gradually caught up with the international level, and finally created in-depth insights and innovations. Eventually, Chinese periodontology has formed a research system with Chinese characteristics and achieved substantial achievements in clinical diagnosis and treatment, basic research, periodontal medicine, and disciplinary integration. Although the current status of Chinese periodontology still lags behind that of developed countries, these representative studies demonstrate the unremitting efforts and hard work of periodontists for generations, laying a solid foundation for the innovation and development of periodontology in our country.

[Distribution and reference intervals of daytime intraocular pressure in the eye health screening population of Handan].

Objective: To describe the distribution and establish reference intervals (RI) of daytime intraocular pressure (IOP) in the eye health screening population of Handan. Methods: This cross-sectional study included subjects who participated in eye health screening at the Physical Examination Center of Handan First Hospital from May 2021 to June 2022. A complete general and ocular examination was performed, including measurements of visual acuity and IOP (using Goldmann tonometry), slit lamp microscopy, fundus photography, and anterior and posterior segment optical coherence tomography. Subjects with factors that could cause significant changes in IOP or affect the accuracy of IOP measurement, or with an inability to measure IOP were excluded. Simple random sampling was used to select participants, who were grouped by gender and age (18 to <30, 30 to <40, 40 to <50, 50 to <60, 60 to <70, and ≥70 years). Central corneal thickness and IOP at 8 to 11 o'clock in one eye of each participant were recorded. The independent sample t test and ANOVA were used for statistical analysis, and the RI of IOP values was calculated by x¯±1.96s. Results: A total of 9 310 subjects had their IOP measured, and 3 491 participants (3 491 eyes) were randomly selected from 7 886 healthy subjects. The age of the participants was (47.74±14.47) years old, ranging from 18 to 90 years old. There were 1 694 males and 1 797 females. The central corneal thickness of all participants was (525.56±49.39) µm. The daytime IOP of all participants was (15.40±2.54) mmHg (1 mmHg=0.133 kPa), and the RI was 10.42 to 20.39 mmHg. The IOP was (15.49±2.58) mmHg for males and (15.29±2.49) mmHg for females, and the gender difference was statistically significant (P<0.05). The RI of daytime IOP values was 10.43 to 20.54 mmHg for males and 10.41 to 20.18 mmHg for females. There were significant differences in daytime IOP [(15.13±2.58), (15.33±2.53), (15.49±2.50), (15.53±2.55), (15.39±2.62), and (15.28±2.52) mmHg] among 6 age groups (P<0.05). Conclusions: The distribution of daytime IOP in different gender and age groups in the eye health screening population of Handan and the RIs derived from the distribution were roughly the same as the international normal IOP RI (10 to 21 mmHg). It is recommended to refer to the RI of daytime IOP values of different genders and ages for clinical decision.

[Urban-rural differentials in the relationship between personality traits and changes in depressive symptoms].

OBJECTIVE: The prevalence of depressive symptoms has become a significant public health issue in China. Research on the relationship between personality traits and changes in depressive symptoms, as well as further exploration of urban-rural differences, not only benefits for the understanding of the prevalence trend of depression in China, but also provides a useful reference for the government to develop personalized mental health prevention strategies. METHODS: Based on the data from the China Family Panel Studies in 2018 and 2020, a univariate analysis was conducted on 16 198 Chinese residents aged 18 years and above. Five dimensions of personality traits were conscientiousness, extraversion, agreeableness, neuroticism and openness. In the study, 16 198 residents were divided into "keep good group", "better group", "worse group" and "keep bad group" according to the changes in depressive symptoms in 2018 and 2020. After controlling for factors, such as gender and education, multinomial Logistic regression analysis was used to examine whether personality traits were associated with changes in depressive symptoms. In addition, we evaluated whether urban-rural and personality traits interacted to influence depressive symptoms. RESULTS: The five dimensions of personality traits were significantly correlated with changes in depressive symptoms. Conscientiousness, extroversion, and agreeableness were negatively associated with depressive symptoms, while neuroticism and openness were positively related. Urban and rural differences moderated the relationship between personality traits and depressive symptoms. Compared with urban residents, rural residents showed stronger correlations between neuroticism (OR=1.14; 95%CI: 1.00-1.30) and the group of depression-recovery, as well as conscientiousness (OR=0.79;95%CI: 0.68-0.93) and the group of persistent-depression. CONCLUSION: The study finds that personality traits have a significant correlation with changes in depressive symptoms, with certain traits showing a negative or positive relationship. Specifically, higher levels of conscientiousness, extraversion, and agreeableness are associated with lower levels of depressive symptoms, while higher levels of neuroticism and openness are associated with higher levels of depressive symptoms. In addition, the study finds that rural residents have a stronger association between their personality traits and persistent or improved depressive symptoms, which highlights the need for tailoring mental health intervention and prevention programs that should take into account personality traits and urban-rural differences in China. By developing targeted strategies that are sensitive to personality differences and geographic disparities, policymakers and mental health professionals can help prevent and reduce the incidence of depressive symptoms, ultimately improving the overall well-being of Chinese adults. Meanwhile, additional studies in independent populations are needed to corroborate the findings of this study.

[Analysis of efficacy and safety of a new endoscopic anastomosis clip in the treatment of defects after endoscopic full-thickness resection].

Objective: To investigate the efficacy and safety of a new endoscopic anastomosis clip in the treatment of defects after endoscopic full-thickness resection (EFTR). Methods: Retrospective cohort study. Fourteen patients [4 males and 10 females, aged (55.9±8.2) years (45-69 years)] with gastric submucosal tumors underwent EFTR at the First Affiliated Hospital of Soochow University were included from December 2018 to January 2021. Patients were divided into new anastomotic clamp group (n=6) and nylon ring combined with metal clips group (n=8). Preoperative endoscopic ultrasound examinations were required to all patients to evaluate the wound condition. The size of the defect, operation time required for wound closure, success rate of closure, postoperative gastric tube placement time, postoperative hospital stay, incidence of complications, preoperative and postoperative serological indexes were compared between the two groups. All patients were followed up after the operation, among which the general endoscopy was reviewed in the first month after the operation, and the telephone and questionnaire follow-up were used in the second, third, sixth month and one year after the operation to evaluate the therapeutic effect of the new endoscopic anastomosis clip and nylon rope combined with metal clip after the EFTR operation. Results: Both groups successfully completed EFTR and were successfully closed. There was no significant difference between the age, tumor diameter and defect diameter of the two groups (all P>0.05). Compared with the nylon ring combined with metal clip group, the operation time of the new anastomotic clip group was shortened [(5.0±1.8) minutes vs (35.6±10.2) minutes, P<0.001]. The operation time was shortened [(62.2±12.5) minutes vs (92.5±0.2) minutes, P=0.007]. Postoperative fasting time decreased [(2.8±0.8) days vs (4.9±1.1) days, P=0.002]. The hospital stay after operation was also shortened [(5.2±0.8) days vs (6.9±1.5) days, P=0.023]. The total intraoperative bleeding volume decreased [(20.00±5.48) ml vs (35.63±14.75) ml, P=0.031]. The patients in both groups received endoscopic examination 1 month after operation, and there was no delayed perforation and bleeding after operation. There was no obvious symptoms of discomfort. Conclusion: The new anastomotic clamp is suitable for the treatment of full-thickness gastric wall defects after EFTR, and shows advantages of shorter operation, less bleeding, and fewer postoperative complications.

The effect of glutathione biosynthesis of Streptococcus thermophilus ST-1 on cocultured Lactobacillus delbrueckii ssp. bulgaricus ATCC11842.

Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus are the main species used for yogurt preparation. Glutathione (GSH) can be synthesized by S. thermophilus and plays a crucial role in combating environmental stress. However, the effect of GSH biosynthesis by S. thermophilus on cocultured L. delbrueckii ssp. bulgaricus is still unknown. In this study, a mutant S. thermophilus ΔgshF was constructed by deleting the GSH synthase. The wild strain S. thermophilus ST-1 and ΔgshF mutants were cocultured with L. delbrueckii ssp. bulgaricus ATCC11842 by using Transwell chambers (Guangzhou Shuopu Biotechnology Co., Ltd.), respectively. It was proven that the GSH synthesized by S. thermophilus ST-1 could be absorbed and used by L. delbrueckii ssp. bulgaricus ATCC11842, and promote growth ability and stress tolerance of L. delbrueckii ssp. bulgaricus ATCC11842. The biomass of L. delbrueckii ssp. bulgaricus ATCC11842 cocultured with S. thermophilus ST-1 or ΔgshF (adding exogenous GSH) increased by 1.8 and 1.4 times compared with the biomass of L. delbrueckii ssp. bulgaricus ATCC11842 cocultured with S. thermophilus ΔgshF. Meanwhile, after H2O2 and low-temperature treatments, the bacterial viability of L. delbrueckii ssp. bulgaricus cocultured with S. thermophilus ΔgshF, with or without GSH, was decreased by 41 and 15% compared with that of L. delbrueckii ssp. bulgaricus cocultured with S. thermophilus ST-1. Furthermore, transcriptome analysis showed that the expression levels of genes involved in purine nucleotide and pyrimidine nucleotide metabolism in L. delbrueckii ssp. bulgaricus ATCC11842 were at least 3 times increased when cocultured with S. thermophilus (fold change > 3.0). Moreover, compared with the mutant strain ΔgshF, the wild-type strain ST-1 could shorten the fermented curd time by 5.3 hours during yogurt preparation. These results indicated that the GSH synthesized by S. thermophilus during cocultivation effectively enhanced the activity of L. delbrueckii ssp. bulgaricus and significantly improved the quality of fermented milk.

[Cough, bloody sputum, eosinophil elevation and lung shadow].

A 47-year-old man was referred to the pulmonary clinic with a 2-year history of productive cough and 3-month history of hemoptysis. Two years ago, his chest CT scan revealed a 2 cm×2 cm well-defined nodule in the right upper lung. His cough was alleviated without treatment. Three months ago, he had a productive cough with the bloody sputum after a running to catch the bus. Physical examination was normal. Complete blood count (CBC) showed an elevated eosinophil count (42.61%). Chest CT scan showed that the enlargement of the right upper lobe nodule (3.4 cm×3.3 cm), with bilateral pathy lesions distributed in the right upper lobe and the left lower lobe. Pathological study of needle specimen biopsy showed the lamellated cyst wall of hydatid cyst, brood capsule formation and hooklet. Pulmonary hydatidosis was diagnosed. It was recommended that the patient should be treated by surgery combined with albendazole. His symptoms relieved and lung nodules were shrinking without treatment after 5 years follow-up.

[Waxing and waning pulmonary nodules and cavities].

We reported a case of vascular Ehlers-Danlos syndrome presenting with recurrent pulmonary hemorrhage. A 22-year-old man was admitted for intermittent hemoptysis and chest pain during the past 18 months. Computed tomography of chest showed bilateral nodules and cavities with halo sign. Inflammatory markers, including erythrocyte sedimentation rate, C reactive protein and interleukin 6, were within normal range. The microbiological and pathological examination of bronchoalveolar lavage fluid and CT-guided percutaneous lung biopsy failed to draw a diagnosis. The pulmonary lesions waxed and waned despite empirical antibacterial, antifungal, antimycobacterial, and anti-parasite treatment. Video-assisted thoracoscopic lung biopsy showed pulmonary hemorrhage, hematoma, ossification, and fibrous nodules, suggesting vascular Ehlers-Danlos syndrome. The molecular testing revealed a heterozygous missense variant in the COL3A1 gene which confirmed the diagnosis of vascular Ehlers-Danlos syndrome. The patient had no skin hyperextensibility or joint hypermobility. During 3-year follow-up, there were no evidence of other vascular or organ involvement except he had intermittent minor hemoptysis. Through this clinical pathological discussion, we aimed to remind pulmonologist to consider the possible diagnosis of vascular Ehlers-Danlos syndrome in young patients with recurrent hemoptysis and waxing and waning pulmonary nodules, cavities, or cysts on CT scan who has neither obvious systematic inflammation nor effective reaction on empirical antimicrobial therapy. Molecular testing should be carried out as soon as possible in a suspected patient to avoid unnecessary invasive examinations.

[Subcutaneous nodules, mediastinal and hilar lymphadenopathy].

A 52-year old man was admitted to our hospital because of dyspnea on exertion for 2 months and subcutaneous nodules for 1 month. Chest enhanced CT showed bilateral hilar and mediastinal lymphadenopathy. Bronchial alveolar lavage fluid revealed a CD4+/CD8+ T cell subsets ratio of 4.3 and culture for acid-fast bacillus (AFB) was negative. The pathology of skin nodules and transbronchial needle aspiration biopsy guided by endoscopic ultrasound (EBUS-TBNA) revealed non-caseating necrotizing epithelioid granulomas with negative acid-fast bacilli staining and periodic acid-Schiff staining, which was compatible with sarcoidosis. The patient was diagnosed as sarcoidosis and glucocorticoid was administrated. The subcutaneous nodules were improved. However, the lymph nodes were enlarged instead of shrinking after 6-month therapy. The failure to respond to glucocorticoids raised the possibility of sarcoidosis complicated with tuberculosis infection. The patient received anti-tuberculosis therapy. Lymphadenopathy responded within 4 months, and there was complete regression after 18-month treatment. The patient was followed up for 5 years and repeated chest CT scan showed that the sizes of bilateral hilar and mediastinal lymph nodes were normal.

[Diagnostic value of surgical lung biopsies for diffuse parenchymal lung disease: the change of disease spectrum in the past 28 years in a single institution in China].

Objective: To investigate the changes of disease spectrum in diffuse parenchymal lung disease (DPLD) diagnosed by surgical lung biopsy, and to explore the diagnostic value of surgical lung biopsy in DPLD. Methods: Four hundred and fifty-five consecutive DPLD patients, who underwent surgical lung biopsy in Peking Union Medical College Hospital during the past 28 years, were analyzed retrospectively. Results: There were 211 males and 244 females. The average age at biopsy was (45±14) years. Four hundred and eleven cases (90.3%) were diagnosed by pathologic findings. Four hundred and forty-one cases (96.9%) were diagnosed by clinical-radiologic-pathologic multidisciplinary discussion. The 30-day mortality and 90-day mortality were 2.4% and 3.3% respectively. The disease spectrum included interstitial pneumonia in 209 cases (45.9%) (nonspecific interstitial pneumonia in 105 cases, usual interstitial pneumonia in 33 cases), other miscellaneous DPLD in 166 cases (36.5%) (including hypersensitivity pneumonitis in 49 cases), tumor in 39 cases (8.6%), and infectious diseases in 27 cases (5.9%). In the three consecutive periods (1993-2002, 2003-2012 and 2013-2020), the number of biopsies was 76 (16.7%), 297 (65.3%) and 82 (18%) respectively. The disease spectrum changes over time: in the above three periods, the percentage of interstitial pneumonia in DPLD was 68.4%, 45.1% and 28%, other miscellaneous DPLDs were 22.4%, 39.4% and 39.0%, the tumors were 2.6%, 7.4% and 18.3%, the infectious diseases were 5.3%, 5.1% and 9.8%. Conclusions: This study presented the changes of disease spectrum in DPLD diagnosed by surgical lung biopsy through single center real-world data, reflecting the progress of clinicians' understanding of DPLD and interstitial pneumonia. Surgical lung biopsy is still valuable for some difficult and complicated DPLD cases.

[Establishment and validation of prognostic risk score model for esophageal squamous cell carcinoma based on immune related genes].

Objective: To find the biomarkers that accurately predict the survival of patients with esophageal squamous cell carcinoma (ESCC). Methods: The immune related genes that were significantly related to the overall survival (OS) of patients with ESCC were screened from The Cancer Genome Atlas (TCGA) database to construct a prognostic risk score model. The prognoses of the high-risk and low-risk groups were compared by Kaplan-Meier method. The accuracy of the model was evaluated by the receiver operating characteristic (ROC) curve. Tumor tissue samples of 83 patients with pathological diagnosis of ESCC were collected from Anyang Cancer Hospital for external verification. Cox regression analysis was used to comprehensively evaluate the effects of prognostic risk score and various clinical characteristics on OS of patients with ESCC. Results: Seven immune-related genes that were significantly related to survival prognosis were selected from the TCGA database and included in the prognostic risk score model, which were S100A12, SLC40A1, FABP9, TNFSF10, IGHA2, IL1F10, and STC2. The 1- and 2-year survival rates of the low-risk group (40 cases) were 94.3% and 82.5%, respectively, while those of the high-risk group (40 cases) were 75.9% and 32.9%, respectively.The prognosis of the high-risk group was worse than that of the low-risk group (P<0.001). The 83 external validation samples obtained consistent results by using the prognostic risk score model. The prognostic risk score was positively correlated with the content of CD4(+) T lymphocytes in ESCC (r(s)=0.259, P=0.020), but not correlated with the content of B lymphocytes, CD8(+) T lymphocytes, neutrophils, macrophages or dendritic cells (P>0.05). Conclusions: S100A12, SLC40A1, FABP9, TNFSF10, IGHA2, IL1F10, and STC2 were risk genes significantly associated with OS of patients with ESCC. The prognostic risk score was an independent prognostic factor for the OS of patients with ESCC, and it was correlated with the content of CD4(+) T lymphocytes in ESCC tissue.

[Hypersensitivity pneumonia diagnosed by excisional lung biopsy: a clinicopathological features of 47 cases].

Objective: To analyze the clinicopathological and immunohistochemical characteristics of the hypersensitivity pneumonia (HP) cases that were diagnosed by excisional lung biopsy, to improve the diagnosis accuracy of HP. Methods: The data of 47 HP cases diagnosed by excisional lung biopsy during the last 20 years were collected in Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, from January 2000 to June 2020. The clinicopathological features and the immunohistochemical profiles of CD3, CD4, CD8 and CD20 were analyzed using light microscopy and immunohistochemical stains. Results: The age of the patients ranged from 15 to 68 years, while the ratio of male to female was 22∶25. The main symptoms were shortness of breath and cough, and 32 cases had a history of allergen exposure. Most of the patients had restrictive impaired ventilatory function, and impaired diffusion function. The proportion of lymphocytes in bronchoalveolar lavage fluid (BALF) increased in 85.3%(29/34) of the patients, and the CD4+/CD8+ T cell ratio was not higher than 0.8 in 31.3%(10/32) of the patients. Microscopically, 59.6%(28/47) of the cases showed nonnecrotizing granuloma/giant cells in the stroma, and 40.4% (19/47) of the cases had only giant cells. Ninety-three-point six percent of the cases(44/47) had peribronchiolar metaplasia, while 70.2%(33/47) had focal cell-rich inflammation. Forty-four cases (93.6%, 44/47) had fibrosis, which was manifested as usual interstitial pneumonia-like fibrosis (47.7%, 21 cases), nonspecific interstitial pneumonia-like fibrosis (29.5%, 13 cases), purely peribronchiolar fibrosis (6.8%, 3 cases) and mixed fibrosis (15.9%, 7 cases). Immunohistochemistry showed that CD4+ T cells were fewer than CD8+ T cells in 24 cases (55.8%, 24/43) of HP. Thirty-two HP patients had follow-up data. Among them, 7 patients had improved symptoms, 12 patients were stable, and 13 patients had worsening symptoms. Conclusions: The most common type of HP diagnosed by excisional lung biopsy is chronic HP. The clinical characteristics are long course of disease, restricted ventilation disorder and impaired diffusion function with the increase of lymphocyte proportion in BALF. Nearly 1/3 of the patients have no known history of allergen contact. The degree of fibrosis in HP patients diagnosed by excisional lung biopsy is severe, and the histologic types of fibrosis varies, but the UIP-like and/or NSIP-like fibrosis is most common. The CD4+/CD8+ T cell ratio in lung tissues is less than 1 in more than half of the cases. Pulmonary fibrosis continued to increase in some of the patients with chronic HP, even after treatment.

Effect and mechanism of miR-217 on drug resistance, invasion and metastasis of ovarian cancer cells through a regulatory axis of CUL4B gene silencing/inhibited Wnt/ß-catenin signaling pathway activation.

OBJECTIVE: To explore the effect and mechanism of miR-217 in cisplatin resistance, as well as invasion and metastasis of ovarian cancer by inhibiting the expression of Cullin 4B (CUL4B) and the activation of Wnt/ß-catenin signaling pathway. MATERIALS AND METHODS: Human ovarian cancer cell lines COC1 (cisplatin sensitive) and COC1/DDP (cisplatin resistant) were cultured and were used to construct the COC1 group and COC1/DDP group, respectively. COC1/DDP cells were divided into blank group, NC group, miR-217 mimic group, miR-217 mimic NC group, miR-217 inhibitor group, miR-217 inhibitor NC group, si-CUL4B group, si-CUL4B NC group, overexpressed (oe) oe-CUL4B group, oe-CUL4B NC group, and miR-217 mimic +oe-CUL4B group, with the identification of cell transfection simultaneously. Bioinformatics prediction and Dual-Luciferase reporter gene assay of the targeting effect of miR-217 on CUL4B were performed, followed by MTT assay for cell proliferation, associated with the measurement of median inhibitory concentration (IC50). Real-time quantitative PCR (qRT-PCR) detected the mRNA expression of miR-217 and CUL4B, and Western blotting for detecting CUL4B, Wnt1, Wnt3, Wnt3a and ß-catenin protein expression. The cell invasion of cells was detected by transwell assay, cell migration by cell scratch assay and cell apoptosis by flow cytometry. RESULTS: Bioinformatics prediction and Dual-Luciferase reporter gene assay verified that CUL4B was a target gene of miR-217, and the latter could silence the expression of the former gene. Compared with COC1 group, the relative expression of miR-217 was significantly decreased, while CUL4B mRNA and protein expression, as well as Wnt1, Wnt3, Wnt3a and ß-catenin protein expression were increased significantly, with evidently increased cell proliferation, IC50, invasion and migration, and decreased apoptosis rate in COC1/DDP group (all p<0.05). Compared with blank group and corresponding NC groups, miR-217 mimic group had increased expression of miR-217 and decreased expression of CUL4B; miR-217 inhibitor group showed decreased miR-217 expression while increased CUL4B expression; si-CUL4B group indicated no significant change of miR-217 expression but decreased CUL4B expression; oe-CUL4B group showed no difference in miR-217 expression but increased CUL4B expression; miR-217 mimic +oe-CUL4B had increased miR-217 expression and no change of CUL4B expression. Meanwhile, miR-217 mimic group and si-CUL4B group exhibited decreased Wnt/ß-catenin, Wnt1, Wnt3, Wnt3a and ß-catenin expression, decreased cell proliferation, IC50, invasion and migration, and increased apoptosis (all p<0.05). Furthermore, miR-217 inhibitor group and oe-CUL4B group revealed increased Wnt/ß-catenin, Wnt1, Wnt3, Wnt3a and ß-catenin expression, increased cell proliferation, IC50, invasion and migration, and decreased apoptosis (all p<0.05). NC group and miR-217 mimic +oe-CUL4B group showed no significant difference in the above indexes (all p>0.05). While compared with miR-217 mimic group, miR-217 mimic +oe-CUL4B group showed increased cell proliferation, IC50, invasion and migration, and decreased apoptosis (all p<0.05). CONCLUSIONS: CUL4B gene is the target gene of miR-217. MiR-217 can silence the expression of this gene, and inhibit the activation of Wnt/ß-catenin signaling pathway to enhance the cisplatin sensitivity and reverse drug resistance, inhibit cell invasion and migration, and promote cell apoptosis.

[Inhibition of the TGF-ß/Smad3 signaling pathway by SB431542: A study of the intervention effect of SB431542 on silicotic fibrosis in rats].

Objective: To investigate the intervention effect of SB431542, which inhibits the TGF-ß/Smad3 signaling pathway, on silicotic fibrosis in rats. Methods: A total of 40 specific pathogen-free Sprague-Dawley rats were divided into normal saline control group, model group, SB431542 inhibitor group, and SB431542 inhibitor control group using a random number table, with 10 rats in each group. All rats except those in the normal saline control group were given non-exposed single intratracheal instillation of free silicon dioxide dust suspension 1 mL (50 mg/mL) ; the rats in the SB431542 inhibitor group were given intraperitoneal injection of SB431542 (5 mg/kg) on days 7 and 30 after dust exposure, those in the SB431542 inhibitor control group were given intraperitoneal injection of SB431542 cosolvent (5 mg/kg) on days 7 and 30 after dust exposure, and those in the normal saline control group were given intratracheal instillation of an equal volume of normal saline (5 mg/kg). On day 60 after dust exposure, the paraffin-embedded section of the right upper lobe of lung was collected for HE staining; the left upper lobe of lung was collected to measure the mRNA levels of fibronectin (FN) , collagen type I (COL-I) , and collagen type III (COL-III) by quantitative real-time PCR; the right inferior lobe of lung was collected to measure the protein levels of FN, COL-I, COL-III, phosphorylated Smad3 (p-Smad3) , and Smad3. Results: Compared with the normal saline control group, the model group had nodules with various sizes in lung tissue, with rupture of some alveolar septa, emphysema changes, and pulmonary interstitial fibrosis, as well as significant increases in the mRNA expression of FN, COL-I, and COL-III and the protein expression of FN, COL-I, COL-III, p-Smad3, and Smad3 in lung tissue (P<0.05) . Compared with the SB431542 inhibitor control group, the SB431542 inhibitor group had a relatively complete structure of lung tissue without marked nodules and with a small amount of exudate in alveolar space and the lumen of bronchioles, as well as significant reductions in the mRNA expression of FN, COL-I, and COL-III and the protein expression of FN, COL-I, COL-III, p-Smad3, and Smad3 in lung tissue (P<0.05) . There were no significant differences in the mRNA expression of FN, COL-I, and COL-III and the protein expression of FN, COL-I, COL-III, p-Smad3, and Smad3 between the model group and the SB431542 inhibitor control group (P>0.05) . Conclusion: SB431542 exerts an intervention effect on silicotic fibrosis by blocking the TGF-ß/Smad3 signaling pathway and reducing the expression of the downstream fibrosis factors FN, COL-I, and COL-III.

Down-regulation of long noncoding RNA DLX6-AS1 defines good prognosis and inhibits proliferation and metastasis in human epithelial ovarian cancer cells via Notch signaling pathway.

OBJECTIVE: Long noncoding RNAs (lncRNAs) are key regulatory RNAs which take(s) part in several biological processes. Recently, a newly identified lncRNA, long noncoding RNA DLX6-AS1 (DLX6-AS1), was reported to be involved in the progression of several tumors. However, its expression and biological function in epithelial ovarian cancer (EOC) have not been investigated. The present study aimed to investigate the role of DLX6-AS1 in the development and progression of EOC. PATIENTS AND METHODS: The expression levels of DLX6-AS1 in EOC tissues and cells were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The association between DLX6-AS1 expression and clinicopathologic characteristics and prognosis in patients with EOC was statistically analyzed. Subsequently, loss-of-function assays were performed to explore alterations in a series of cells phenotypes, including cell viability, colony formation, cell cycle, apoptosis, colony formation, and migration and invasion capacities. The effect of DLX6-AS1 on the Notch signaling pathway was evaluated by Western blot and RT-PCR. RESULTS: We first verified the increased expression of DLX6-AS1 in EOC patient samples and cell lines. Clinical assays indicated that high DLX6-AS1 was significantly associated with FIGO stage, lymph node metastasis and poor prognosis. Furthermore, multivariate Cox regression analysis confirmed DLX6-AS1 as an independent prognostic factor in EOC patients. Functionally, the down-regulation of DLX6-AS1 decreased EOC cells proliferation, migration, invasion and induced cell cycle G1/S phase arrest and cell apoptosis. Mechanistic studies revealed that the knockdown of DLX6-AS1 down-regulated Notch1, p21, and Hes1, indicating that the activity of the Notch signaling pathway was inhibited. CONCLUSIONS: Our results demonstrated that DLX6-AS1 inhibited a tumor-promoting role in EOC and may be useful as a prognostic biomarker and/or a therapeutic avenue for EOC.

[Preliminary study on the measurement of optic nerve subarachnoid space in patients with normal tension glaucoma].

Objective: To investigate the ultrasonographic measurements of the optic nerve subarachnoid space (ONSAS) in patients with normal tension glaucoma (NTG). Methods: Cross-sectional study. Patients with NTG or primary open-angle glaucoma (POAG) but without any anti-glaucoma treatment and the control group were collected from June 2016 to March 2017 at Beijing Tongren Hospital affiliated to Capital Medical University. Measurements of 24-h intraocular pressure, central corneal thickness, mean visual field damage, visual axis, blood pressure and body mass index and ocular ultrasound scans were performed. The differences in the ONSAS of the two-dimensional ultrasound images of the three groups of subjects and their correlation with various clinical variables were evaluated. Qualitative data were analyzed by the chi-square test for comparison between groups; quantitative data were analyzed by the one-way ANOVA for comparison between groups, and the LSD-t test was used for comparison between groups; the Pearson correlation analysis was used for correlation analysis of measurement data. The intra-group correlation coefficient was used to evaluate the reproducibility and repeatability of two measurements by different measurers or the same measurer. Results: A total of 35 patients (35 eyes) with NTG were enrolled in the NTG group, including 14 males and 21 females, aged (50±8) years; 32 patients (32 eyes) with POAG were enrolled in the POAG group, including 14 males and 18 females, aged (52±10) years; 37 healthy people (37 eyes) were enrolled in the control group, including 20 males and 17 females, aged (49±9) years. The ONSAS was (5.07±0.83) mm(2) in the retrobulbar 3- to 7-mm range in the NTG group, significantly smaller than the control group (6.57±1.43) mm(2) and the POAG group (6.19±0.90) mm(2) (t=1.17, 1.29; P=0.03, 0.01). There was no significant difference in the statistical results between the control group and the POAG group (t=1.31, P=0.75). Between the ONSAS and mean intraocular pressure and maximum intraocular pressure, there was a statistically positive correlation in the NTG group (r=0.66, 0.48; both P<0.01), but there was no linear correlation; there was no statistical correlation in the control group or the POAG group (all P>0.05). There was no statistically significant association between the ONSAS and age, mean arterial pressure, body mass index, central corneal thickness, visual axis length, and mean visual field loss in any group (all P>0.05). Conclusion: Compared with the POAG group and the control group, the ONSAS is smaller in the NTG group, indicating lower retrobulbar cerebrospinal fluid pressure, and it is positively correlated with the mean intraocular pressure and the highest intraocular pressure. (Chin J Ophthalmol, 2018, 54: 827-832).

[Experimental studies on correlation between nucleolus spindle-related protein 1 and the malignant progression and prognosis of human glioblastoma multiforme].

Objective: To investigate the correlation between nucleolus spindle-related protein 1 (NUSAP1) and malignant progression and prognosis of human glioblastoma multiforme (GBM). Methods: RT-PCR and immunohistochemical technique were applied to analyze NUSAP1 expression level in GBM surgical specimens. Correlations between NUSAP1 expression and molecular classification and survival of patients with GBM were also investigated in TCGA database. The gene silencing technique was used to silence NUSAP1 expression in U87 cells, CCK-8 assay was used to detect cell proliferation, flow cytometry was used to detect cell cycle changes, and in vivo tumorigenicity was evaluated after NUSAP1 silencing in tumor-bearing mice. Results: NUSAP1 expression level in GBM was higher than that in non-tumor brain tissue. Survival curve analysis showed that the survival time of GBM patients with high NUSAP1 expression decreased significantly (P<0.01). NUSAP1 expression was relatively lower in mesenchymal and neural molecular subtypes of GBM, when compared with the other two molecular subtypes. And it was closely related with specific genetic aberrations (such as PTEN loss and IDH1 mutation). Silencing NUSAP1 inhibited G2/M cell cycle progression of GBM cells, and inhibited cell proliferation both in vitro and in vivo. Conclusion: Expression of NUSAP1 is closely related to progress and prognosis of GBM, and can be a biomarker reflecting GBM prognosis and act as a therapeutic target with potential clinical application value.

Increase in G protein-coupled receptor autoantibodies with decline of cardiac function in hypercholesterolemic rats.

OBJECTIVE: This study aimed to explore the effects of diet-induced hypercholesterolemia (HC) on the production of G protein-coupled receptor autoantibodies and to elucidate the potential mechanisms involved. MATERIALS AND METHODS: Male Wistar rats were fed a normal or high-cholesterol diet for 8 weeks. Cardiac function, autoantibodies against G protein-coupled receptors, the beat frequency of neonatal cardiomyocytes, the CD4+/CD8+ T-lymphocyte ratio and lymph leukocyte counts in the spleen were determined. RESULTS: Diet-induced hypercholesterolemia significantly increased the levels of autoantibodies against α1- and ß1-adrenergic receptors and the angiotensin II type 1 receptor in sera, as well as the CD4+/CD8+ T-lymphocyte ratio and lymph leukocyte count in the spleen, and decreased cardiac function. There were strong negative correlations between the levels of autoantibodies and cardiac injury. CONCLUSIONS: The present study demonstrates, for the first time, that G protein-coupled receptor autoantibodies exist in the sera of hypercholesterolemic rats and that the levels of these autoantibodies are related to cardiac function, which implies that these cardiac receptor autoantibodies may play a role in cardiac dysfunction in hypercholesterolemic rats.

[Down-regulation of miR-146b-5p promotes malignant transformation of fusion cells after co-culture of macrophages with glioma stem cells in vitro].

Objective: To observe mutual interactions between macrophages(Mφ) and glioma stem cells (GSCs)in dual-color tracing model in vitro, to identify the biological characteristics of fusion cells in multiple levels, and to analysis the relevant molecular mechanisms. Methods: Red fluorescent protein(RFP) gene was stably transfected into human GSCs cell line SU4. Mφ cells were obtained from Balb/c nude mice with enhanced green fluorescent protein (EGFP) expression. Then two cells were co-cultured in dual-color tracing platform. RFP/EGFP double positive cells with high proliferation ability were mono-cloned. The fusion cells were verified by Western blot, fluorescence in situ hybridization, immunocytochemistry and chromosome karyotype analysis.The biological characteristics of fusion cells were further analyzed, together with relevant molecular changes. Results: RFP / EGFP double positive cells were obtained through in vitro co-culture. RFP and EGFP coexpression were proved at transcriptional and translational levels in the fusion cells. They also co-expressed GSCs marker Nestin and Mφ marker CD68, and karyotype analysis showed two types of characteristic chromosomes, which confirmed that the fusion cells originated from spontaneous fusion between SU4-RFP and Mφ.Fusion cell proliferation rate and invasion ability were higher than SU4-RFP, which were relevant with down-regulation of miR-146b-5p and activation of STAT3. Fusion cells transfected with miR-146b-5p showed a higher apoptosis rate(18.83%) and lower tumor formation(4/5). Conclusion: Mφ could fuse with GSCs spontaneously in local tumor micro-environment. The proliferation and invasion abilities of fusion cells were higher than their parent cells, which were relevant with down-regulation of miR-146b-5p and activation of STAT3. It revealed the possible mechanisms of malignant progression of gliomas.