[A case of acute exudative polymorphous vitelliform maculopathy].

A 35-year-old male patient presented to the hospital with binocular blurred vision for 2 weeks. The visual acuity of both eyes was 0.8. Fundus examination showed multiple yellow-white punctate lesions in the posterior pole of both eyes. OCT showed cystoid edema and submacular edema, thickening of ellipsoid zone and enhancement of reflex in macular region. Fundus autofluorescence showed strong autofluorescence at the lesion site. Fundus fluorescein angiography showed no fluorescence leakage in the lesion area. The patient was diagnosed with acute exudative polymorphous vitelliform maculopathy based on medical history, ocular multimodal examination and general examination. The patient was not given special treatment, but one week later, the lesion was fused and expanded, and the macular edema was worse than before.

[Consideration on the construction of a special research database for human papillomavirus vaccination in high-risk and special populations].

The application of human papillomavirus (HPV) vaccine in China has accelerated in recent year. Women's and Children's hospitals as well as general hospitals have established HPV vaccination counselling clinic in succession. A large amount of valuable preventive and clinical data have been generated in the evaluation, recommendation, immunization and management of HPV vaccine for high-risk and special populations. This article sorted out the operation process of HPV vaccination counselling clinic, discussed the methods for hospitals to construct a special research database for HPV vaccination in high-risk and special populations under different degrees of informatization. It also provided a reference for the establishment of unified data standards, the formation of available data resources, and the promotion of real world study on HPV vaccination for high-risk and special populations in China.

[Enterostomy based on abdominal wall tension and fascial locking: a theory of preventing stoma complications and parahernia].

No consensus on standardized technique of enterostomy creation has been made meanwhile high heterogeneity of surgical procedure exists in 'stoma creation' chapters of textbooks or atlases of colorectal surgery. The present article reviews the anatomy of tendinous aponeurotic fibers which is crucial for abdominal wall tension and integrity. Through empirical practice we hypothesize a procedure of enterostomy creation basied on abdominal wall tension plus anchor suture for fascia fixation which could theoretically decrease short-term stoma complication rates and long-term parastomal hernia rates. Surgical techniques are as followed: (1) preoperative stoma site mark for de-functioning ileostomy should be positioned at the lateral border of rectus abdominis muscle (RAM) to decrease the difficulty of stoma reversal and for permanent colostomy should be placed overlying the RAM to promote adhesion; (2)Optimal circular removal or lineal opening of skin, and avoid dissection of subcutaneous tissue; (3) Lineal dissection of natural strong fascia (rectus sheath) at stoma site and blunt separation of muscular fibers. The tunnel of the fascia should be made with appropriate size without undue tension. To prevent the formation of dead space, additional suturing at fascia layer is unnecessary. (4) Anchor suture for fascia fixation at two ends of fascia opening could be considered to avoid delayed fascia disruption and parastomal hernia. (5) After pull-through of ileum or colon loop, 4-8 interrupted seromuscular sutures could be placed to attach loop to skin. For ileostomy, self-eversion of mucosa can be successful in vast majority of cases and a Brooke ileostomy is not necessary. The efficacy and safety of this procedure should be tested in future trials.

[Screening and analysis of prognostic factors of repairing single missing tooth by autotransplantation of teeth].

Objective: To screen and analyze the factors affecting the prognosis of replacing single missing tooth by autograft tooth, so as to provide reference for clinical judgment of surgical prognosis. Methods: A total of 176 patients (188 teeth) underwent autotransplantation of teeth in the Department of Oral & Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University from January 2017 to December 2019, including 85 teeth of males and 103 teeth of females were involved. The age was (33.0±9.8) years (16-65 years). The possible factors affecting the prognosis of replacing single missing tooth by autograft tooth were summarized and grouped, and the clinical and imaging data were recorded and judged. The surgical records and photographic data from the patients' previous medical records were retrospectively analyzed. The survival analysis method was used for statistical analysis to screen out the factors affecting the cumulative survival rate of transplanted teeth. Results: The 5-year cumulative survival rate of 188 transplanted teeth was 88.4%. Univariate Log-Rank analysis showed that age (P<0.001), sex (P=0.008), smoking (P<0.001), position of recipient area (P<0.001), height of alveolar bone in recipient area (P<0.001), time of donor tooth in vitro (P<0.001), use of donor model (P<0.001) and initial stability (P<0.001) were significantly correlated with cumulative survival rate of transplanted teeth. Multivariate Cox proportional hazard regression analysis showed that smoking (ß=-2.812, P=0.049), alveolar bone height (ß=1.521, P=0.020), donor time (ß=-2.001, P=0.019), use of donor model (ß=1.666, P=0.034) and initial stability (ß=-1.417, P=0.033) were significantly correlated with the cumulative survival rate of transplanted teeth. Conclusions: The prognosis of autogenous tooth transplantation can be predicted by smoking, height of alveolar bone in recipient area, time of donor teeth in vitro, use of donor model and initial stability. Good prognosis of transplanted teeth can be obtained by using donor model during operation, reducing the time of donor teeth in vitro, taking effective methods to restore alveolar bone height, maintaining good initial stability, and good oral health education after operation.

[Diffuse midline gliomas with H3K27 alteration in children: a clinicopathological analysis of forty-one cases].

Objective: To investigate the clinicopathological features of pediatric diffuse midline glioma with H3K27 alteration and to analyze their relationship with prognosis. Methods: Forty-one cases of childhood diffuse midline glioma with H3K27 alteration were collected at Children's Hospital of Fudan University (39 cases) and Xi'an Children's Hospital (2 cases), from July 2016 to July 2020. The clinical manifestations, imaging data, histopathology, immunohistochemical phenotype and molecular genetics features, tumor size, site and histological grading were evaluated. Results: Among the 41 cases, 21 were males and 20 females, the age of onset was 3-14 years, the average and median age was 7.6 years and 7.0 years, respectively. The tumor sites were brain stem (n=36) and other locations (n=5). The clinical manifestations were dizziness, gait disturbance, and limb weakness, etc. The MRI features were variable. The histology varied from low-grade to high-grade glioma with neuron differentiation. Immunohistochemistry showed that the tumor cells expressed H3K27M, GFAP, and Olig2. Genetic study showed that 76% (16/21) of tumors had H3F3A gene mutation, mostly accompanied by TP53 (62%, 13/21) missense mutation; five tumors (24%, 5/21) had HIST1H3B gene mutation, accompanied by missense mutations in ACVR1 and PI3K pathway-related gene PIK3CA (4/5) and PIK3R1 (1/5) mutations. The prognosis was dismal with only one alive and others died. The average and median overall survival time was 7 months and 4 months, respectively. Cox multivariate regression analysis showed that age, tumor location, radiologically maximum tumor diameter, histologic grading, and surgical methods were not significantly associated with overall survival rate (P>0.05). Conclusions: Pediatric diffuse midline gliomas with H3K27 alteration have unique clinicopathological and genetic characteristics. The prognosis is poor. The tumor location and histopathologic grading are not related to prognosis. New specific drugs and comprehensive treatment are needed to improve the prognosis.

[Classification of tracheobronchial tuberculosis in 252 children].

Objective: To provide theoretical basis for early diagnosis and accurate bronchoscopic classification of tracheobronchial tuberculosis (TBTB) in children through analyzing the clinical characteristics, bronchoscopic classifications and treatment effect in children with TBTB. Methods: In this respective study, we collected clinical data of patients with TBTB who accepted bronchoscopies in Interventional Pulmonology Department of Beijing Children's Hospital Affiliated to Capital Medical University between January, 2006 and December, 2019. The basic data, including clinical manifestations, imaging features, bronchoscopic characteristics and effects of interventional therapy were analyzed. The results of the study were statistically described and analyzed using SPSS 22.0 statistical software for relevant data. Results: Total 252 children with TBTB were included in this study. The median age was 1.7 years (quartile: 0.8 years, 5.2 years). Analysis of the classification of TBTB showed that the percent of lymph node fistula type was 96.4% (243/252), ulcerative necrosis type 1.2%(3/252), granulation proliferation type 0.4% (1/252), and cicatricial stricture type 0.8% (2/252). In addition, 1.2% (3/252) of the cases showed the same bronchoscopic manifestations as lymph node fistula type, but it was not clear on imaging whether the caseous material in the lumen was caused by lymph node or lung erosion. Therefore, the "bronchial fistula type" was proposed. Conclusions: Lymph node fistula type of TBTB was the common in children. The classification of lymph node fistula mostly depended on imaging evidence, and this may lead to some uncertainty in classifying TBTB in cases with no imaging evidence of enlarged lymph nodes.

[Effect of Echinococcus multilocularis infections on mitochondrial functions of macrophages].

OBJECTIVE: To investigate the changes of mitochondrial metabolic functions of macrophages following Echinococcus multilocularis infections, so as to provide insights into the pathogenesis of alveolar echinococcosis. METHODS: Two groups were assigned according to different treatment methods. In the culture group, mouse leukemic monocyte macrophage RAW264.7 cells were cultured with 2 000 E. multilocularis at a ratio of 500∶1, while RAW264.7 cells in the control group were given no treatment. Then, both the culture and control groups were further divided into the 24 h and 72 h subgroups. Mitochondria were stained with MitoTracker® Deep Red FM and the mean fluorescence intensity of macrophage mitochondria was measured with the Cytation 5 Cell Imaging Multi-Mode Reader. The mitochondrial DNA copy number was quantified using the quantitative real-time PCR (qPCR) assay, and the mitochondrial energy metabolism was monitored using the Seahorse XF assay. In addition, the mitochondrial reactive oxygen species and mitochondrial membrane potential were detected using flow cytometry. RESULTS: The mean fluorescence intensities of macrophage mitochondria were significantly lower in the 24 h (15 341 ± 2 532 vs. 17 823 ± 3 429; t = 6.379, P < 0.01) and 72 h (18 102 ± 3 505 vs. 21 511 ± 5 144; t = 17.680, P < 0.01) culture subgroups than in the corresponding control subgroups, and lower mitochondrial DNA copy numbers were measured in the 72 h culture subgroup than in the 72 h control group [(3.23 × 109 ± 1.78 × 107) vs. (4.39 × 109 ± 3.70 × 107); t = 8.85, P < 0.001]. The oxygen consumption rates were significantly greater in the 24 h [(241.70 ± 73.13) pmol/min vs. (69.05 ± 52.30) pmol/min; t = 7.89, P < 0.01] and 48 h culture groups [(249.50 ± 42.06) pmol/min vs. (60.28 ± 40.66) pmol/min; t = 8.64, P < 0.01] than in the corresponding control groups, and a higher extracellular acidification rate was seen in the 48 h culture group than in the 48 h control group ([ 111.6 ± 17.49) mpH/min vs. (35.05 ± 7.57) mpH/min; t = 16.90, P < 0.01]. In addition, flow cytometry detected higher mean fluorescence intensity of mitochondrial reactive oxygen species (58 264 ± 10 087 vs. 4 307 ± 97; t = 12.930, P < 0.01) and lower mitochondrial membrane potential (9.833% ± 2.285% vs. 2.667% ± 0.208%; t = 6.645, P < 0.01) in the 72 h culture group than in the control group. CONCLUSIONS: E. multilocularis infection may impair mitochondrial functions and inhibit oxidative phosphorylation of macrophages, resulting in increased macrophage glycolysis. It is speculated that the alteration of macrophage metabolic states may contribute to the mechanisms underlying the development and progression of alveolar echinococcosis.

[Analysis of the relationship between the anatomical location of intrapulmonary metastatic lymph nodes and relapse risk and survival in patients with N1 non-small cell lung cancer].

Objective: To evaluate the relationship between the anatomical location of intrapulmonary metastatic lymph nodes and relapse risk and survival in patients with N1 non-small cell lung cancer(NSCLC). Methods: A retrospective analysis of the clinical and pathological data of 138 patients with completely resected N1 NSCLC was conducted. There were 79 males and 59 females, aged from 26 to 81 years with an average of (59±10) years. All of them were treated in the Department of Thoracic Surgery Ⅱ of Peking University Cancer Hospital between January 2007 and December 2015. Patients were stratified based on the 8th edition of the American Joint Committee on Cancer (AJCC) N1 classification and the modified pathological N1 classification strategy, respectively. According to modified pathological N1 classification strategy, which was defined based on the anatomical location of intrapulmonary metastatic lymph nodes, N1 nodes were subcategorized into the hilar (stations 10-11, mN1b) (n=36) and peripheral (stations 12-14, mN1a) (n=102) zones. The Kaplan-Meier curves were plotted to compare the relapse risk and survival analysis, disease-free survival (DFS), and overall survival (OS) were compared between the two staging methods through univariate and multivariate analysis to evaluate the effectiveness of the two classifications in stratifying patients with distinct risks of disease relapse and survival. Results: According to the modified N1 classification, the differences in 5-year DFS and OS between the subgroups (mN1a vs mN1b) were statistically significant(59.5% vs 35.7%; 81.2% vs 56.0%; both P<0.05). However, following the 8th edition of the AJCC N1 classification, no significant differences were found in DFS and OS between the subgroups (both P>0.05). Multivariate analysis showed that the modified N1 classification was an independent prognostic factor to DFS (HR=1.814, 95%CI: 1.005-3.275) and OS (HR=3.919, 95%CI: 1.918-8.009) (all P<0.05). However, the 8th edition of the AJCC N1 classification was not an independent prognostic factor to DFS (HR=1.360, 95%CI:0.767-2.412) or OS (HR=1.620, 95%CI:0.839-3.131) (both P>0.05) as revealed by multivariate analysis. Conclusions: The relapse risk and survival could be assessed effectively using the modified pathological N1 classification, which was defined and subcategorized based on the anatomical location of intrapulmonary metastatic lymph nodes for N1 NSCLC patients. The modified pathological N1 classification is superior to the 8th edition of the AJCC classification.

[Mutation analysis and prenatal diagnosis of MYO7A gene in a case of Usher syndrome type 1].

Objective: To analyze the clinical characteristics and identify the causative gene of a case with congenital deafness. Methods: Detailed medical history and clinical examination of a 4-year-old male child with congenital deafness were conducted in the First Affiliated Hospital of Army Military Medical University in June 2016. He was diagnosed with sensorineural deafness. The venous blood of the child and his parents was drawn, and genomic DNA was extracted. Proband's DNA was performed with targeted capture of high-throughput sequencing, then Sanger sequencing was used to verify the suspected mutation and segregation in this pedigree. According to the genetic diagnosis of the proband's deafness, ophthalmic examinations were performed. Genetic prenatal diagnosis was performed when the proband's mother was pregnant again. Results: The patient was detected with p.Trp1466Ter/p.Tyr2042Ter compound heterozygous mutations of MYO7A gene with targeted high-throughput sequencing. The mutation of p.Trp1466Ter was a reported mutation, while p.Tyr2042Ter has not been reported. In addition to congenital deafness, retinitis pigmentosa was also found by ophthalmologic examination, and the patient was clinically diagnosed with Usher syndrome type 1. Amniocentesis and fetal DNA sequencing were performed on the repregnancy fetus of this family at 18 weeks of gestation. The heterozygous mutation of MYO7A gene p.Tyr2042Ter was found, and the other allele was the wild type, indicating that the child will not exhibit clinical manifestations of Usher syndrome type 1. Indeed, the second child passed neonatal hearing screening. Conclusions: The clinical features and genetic variants were delineated in this family with Usher syndrome type 1. The results of the current study have enriched the phenotype and genotype data of the disease and provided a basis for genetic counseling.

[Effects of stomatin protein expression on proliferation and apoptosis of lung cancer cells].

Objective: To investigate the effect of stomatin protein expression on the proliferation and apoptosis of lung cancer cells. Methods: The expressions of stomatin mRNA in human bronchial epithelial cells (HBE) and lung cancer cells (H520, A549, 95D, H460, Glc-82, 973 and H1299) were detected by Real-time PCR. Immunohistochemistry (IHC) was used to detect stomatin protein expression in 4 lung cancer tissue microarrays with 259 cases of lung cancer and adjacent normal tissues. After knocking down the expression of stomatin in A549 cells, the proliferation was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, the apoptosis was detected by flow cytometry, the expression levels of total protein kinase B (AKT) and phosphorylated AKT at Ser473 were detected by Western blot. BALB/c nude mice were used to detect the tumorigenic ability of stomatin downregulated A549 cells (3 mice) and control cells (3 mice), and the protein expressions of stomatin, Ki67 and CD31 in tumor tissues were detected by IHC. Results: The M (range) of stomatin mRNA expression level in H520, A549, 95D, H460, Glc-82, 973, H1299 and HBE cells were 2.71 (2.66), 3.55 (3.16), 0.26 (0.22), 2.08 (1.98), 0.87 (0.35), 1.72 (2.53), 1.10 (1.82) and 0.01 (0.02), respectively. The mRNA expression levels of stomatin in H520, A549 and H460 cells were higher than that of HBE cells (all P<0.05), whereas there was no statistical difference among 95D, Glc-82, 973, H1299 and HBE cells (all P>0.05). IHC of lung tissue microarrays showed that the positive rate of stomatin expression in human lung cancer tissues was 34.7% (90/259), which was significantly higher than that in adjacent normal tissues [1.9% (5/259)] (P<0.05). In stomatin positive lung cancer tissues, the M (IQR) of tumor size for lower stomatin expression tissues (67 cases) was [41.22 (2 761.50) cm], which was smaller than that of higher stomatin expression tissues [(23 cases, 57.98(1 333.50) cm) (P<0.05). After knocking down stomatin expression, the fourth day absorbance value of stomatin-downregulated A549 cells was 0.55±0.07, which was lower than that of control cells (0.79±0.16) (P=0.012). The proportion of early apoptotic cells of stomatin-downregulated A549 cells [8.83 (53.00)] was higher than that of control cells [4.17 (25.00)] (P=0.026). The Ser473 phosphorylated AKT protein expression level in stomatin-downregulated A549 cells was 0.68±0.16, which was decreased compared with control cells (1.16±0.39) (P<0.05). The M (IQR) of total AKT expression level in stomatin-downregulated A549 cells was 4.25 (17.00), without statistically significant difference from control cells [4.75 (19.00)] (P>0.05). After inoculation of stomatin-downregulated A549 cells in nude mice for 43 days, the tumor volume was (37.93±3.12) mm(3), which was significantly smaller than that of the control group [(454.04±32.39) mm(3)] (P<0.001). And the expression levels of stomatin, nuclear proliferation antigen Ki67, and platelet-endothelial cell adhesion molecule CD31 were 1.78±0.69, 5.19±3.84, and 10.77±1.67, respectively, which were all decreased compared with control group (17.52±8.76, 54.14±41.02, and 19.72±6.97, respectively) (all P<0.05). Conclusion: Stomatin promotes lung cancer cell proliferation and inhibits cell early apoptosis by regulating AKT signaling pathway.

OIP5-AS1 promotes the progression of gastric cancer cells via the miR-153-3p/ZBTB2 axis.

OBJECTIVE: Gastric cancer (GC) remains a serious disease to human health with high mortality worldwide. Evolving evidence implied that long non-coding RNA Opa interacting protein 5-antisense RNA1 (OIP5-AS1) went in for the pathological progress of GC. Nevertheless, the potential molecular mechanism of OIP5-AS1 needed to be further investigated. MATERIALS AND METHODS: Levels of OIP5-AS1, microRNA (miR)-153-3p, and zinc finger and BTB domain containing 2 (ZBTB2) were assessed using quantitative real-time polymerase chain reaction (qRT-PCR) or Western blot assays. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) was implemented to detect cell proliferation in vitro. Cell apoptosis was evaluated by flow cytometry. Besides, transwell assay was conducted to examine cell migration and invasion in AGS and MKN45 cells. The interaction between miR-153-3p and OIP5-AS1 or ZBTB2 was validated utilizing dual-luciferase reporter assay. Lastly, the role of OIP5-AS1 in tumor growth was researched through adopting xenograft tumor model. RESULTS: OIP5-AS1 and ZBTB2 were strongly higher in GC tissues than noncancerous samples. OIP5-AS1 silencing remarkably curbed cell proliferation, migration and invasion, and elevated cell apoptosis in both AGS and MKN45 cells. Functional analysis indicated that OIP5-AS1 regulated ZBTB2 expression via binding to miR-153-3p. Moreover, the role of miR-153-3p in cell growth and metastasis was abrogated by ZBTB2 overexpression. Above all, OIP5-AS1 could reduce the growth of xenograft tumor in vivo. CONCLUSIONS: OIP5-AS1 exerted its role via miR-153-3p/ZBTB2 axis in the progression of GC cells. These findings might supply a biomarker for the diagnosis and therapy of GC clinically.

[Expression of p-AKT and p-mTOR in pediatric Burkitt lymphoma and their correlation with prognosis].

Objective: To evaluate the expression of p-AKT and p-mTOR, the key proteins in PI3K/AKT/mTOR pathway in pediatric Burkitt lymphoma (BL), and to investigate the clinical and prognostic significance. Methods: Fifty-eight cases of pediatric BL and thirty cases of reactive hyperplastic lymphadenitis (RH) were collected at Children's Hospital of Fudan University from September 2011 to July 2018. Paraffin sections of tissues were immune stained for p-AKT and p-mTOR, and the expression was assessed and correlated with the clinical features and prognosis. Results: A total of 58 cases were diagnosed and 6 cases lost the follow-up. Of the remaining 52 BL patients including 43 males and 9 females, the median age was 5 years (range: 2 to 14 years). Regarding to the correlation between the two biomarkers, Spearman test showed that p-mTOR was positively associated with the expression of p-AKT (r=0.759, P<0.001). Of all BL patients, the positive rates of p-AKT and p-mTOR were 62.1% (36/58) and 60.3%(35/58) respectively, both significantly higher than control group (P=0.011, P=0.035 respectively). The presence of p-AKT was significantly associated with higher lactate dehydrogenase (LDH≥573 IU/L) level in patients of the disease (P=0.006), while p-mTOR was increased both in the higher LDH and lower ratio of albumin to globulin (A/G) group (P=0.006, P=0.034 respectively). Expression of p-AKT and p-mTOR did not show any statistical correlation with sex, age, St.jude stage, tumor size, B-symptom present or not, number of extra-nodal sites or international prognostic index (IPI) (P>0.05). Fifty-two patients had a median follow-up of 40 months (range: 5-87 months). Univariate analysis showed that p-AKT expression was significant in predicting both inferior OS (5-year estimate, 72.7% vs. 94.7%, χ(2)=4.123, P=0.042) and PFS (5-year estimate, 66.7% vs. 94.7%, χ(2)=5.822, P=0.016). The 5-year OS rate was 71.0% (22/31) for the p-mTOR positive cohort of patients compared to 95.2% (17/21) for p-mTOR negative group (χ(2)=4.881, P=0.027); however, there was no statistical significance in 5-year PFS rate (P>0.05). Especially, the 5-year OS and PFS rate of p-AKT/p-mTOR double-positive group were significantly lower than negative control group (including absence of single p-AKT or p-mTOR expression, and absence of both) (OS: 69.0% vs. 95.7%, χ(2)=6.285, P=0.012; PFS: 65.5% vs. 91.3%, χ(2)=5.405, P=0.020). The results of multivariate COX proportional risk regression analysis indicated that p-AKT/p-mTOR double-positive, higher LDH and IPI score 3-5 were independent prognostic factors for both OS and PFS, and the bulky tumor (>10 cm) for PFS of pediatric BL. Conclusion: The expression of p-AKT and p-mTOR may be a potential reference for diagnosis and the independent prognostic indicators of pediatric BL.

Effect of miR-133 on apoptosis of trophoblasts in human placenta tissues via Rho/ROCK signaling pathway.

OBJECTIVE: The aim of this study was to explore the role of micro ribonucleic acid (miR)-133 in the apoptosis of human placental trophoblasts through the Ras homolog gene family (Rho)/Rho-associated coiled-coil forming protein kinase (ROCK) signaling pathway. PATIENTS AND METHODS: The plasma samples were collected from 30 patients with pre-eclampsia (PE) undergoing treatment and 30 healthy subjects (control group) who received physical examination in our hospital. The Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was utilized to measure the expression of miR-133 in PE patients and healthy people. Meanwhile, blood pressure, urine protein content, liver function, and kidney function were detected in patients of both groups as well. Subsequently, the placental trophoblasts were extracted and transfected with inhibitors and miRNA mimics to suppress and overexpress miR-133, respectively. The transfection efficiency was determined by RT-PCR. The levels of interleukin-6 (IL-6), IL-1, and tumor necrosis factor-alpha (TNF-α) were measured in both groups. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assay was performed to determine the apoptosis of trophoblasts. Next, the RT-PCR and Western blotting were carried out to detect the expressions of the Rho/ROCK pathway. Furthermore, the influence of miR-133 on the apoptosis of trophoblasts in human placenta tissues through Rho/ROCK was comprehensively observed. RESULTS: In vivo experiments demonstrated that the urinary protein content, miR-133 level, systolic blood pressure, diastolic blood pressure, and liver function and renal function indexes were significantly elevated in pre-eclampsia (PE) patients in comparison with normal subjects (p<0.05). After transfection of mimics and inhibitors, the expression of miR-133 was remarkably up- and down-regulated, respectively. The content of the inflammatory factors in miR-133 mimics group was overtly higher than the other two groups. The TUNEL staining results showed that the number of apoptotic cells significantly increased and decreased in the miR-133 mimics group and miR-133 inhibitors group, respectively. Subsequent experiments indicated that the expressions of apoptosis gene Caspase3, pathway gene, and protein ROCKI were notably up-regulated in miR-133 mimics group. However, they were evidently down-regulated in miR-133 inhibitors group than in the control group. In addition, a consistent trend was observed in the protein expression level. CONCLUSIONS: MiR-133 participates in the development and progression of PE through the Rho/ROCK signaling pathway, which may affect the apoptosis of trophoblasts in the placenta tissues.

Long non-coding RNA TOB1-AS1 modulates cell proliferation, apoptosis, migration and invasion through miR-23a/NEU1 axis via Wnt/b-catenin pathway in gastric cancer.

OBJECTIVE: Gastric cancer (GC) is the fourth common cancer worldwide. Long non-coding RNA TOB1 antisense RNA 1 (TOB1-AS1) has been found to participate in the process of GC, while the precise role of TOB1-AS1 is still not understood in GC progression. MATERIALS AND METHODS: We collected 21-paired GC and para-carcinoma tissue specimens, and the levels of TOB1-AS1 and lysosomal sialidase (NEU1) were detected by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). The protein expression levels of NEU1, b-catenin, c-Myc, Cyclin D1, N-cadherin were determined via Western blot. Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry was performed to evaluate cell proliferation. Besides, GC cell migration and invasion capacities were identified by transwell assay. Dual-Luciferase reporter assay was employed to examine the interrelation between miR-23a and TOB1-AS1 or NEU1. Finally, the role of TOB1-AS1 was verified in vivo. RESULTS: The levels of TOB1-AS1 were decreased in GC tissues and cell lines. Either TOB1-AS1 or NEU1 upregulation accelerated GC cell apoptosis, hampered proliferation, migration, and invasion. Further, the role of TOB1-AS1 silencing on cell behaviors was abrogated by NEU1 upregulation. TOB1-AS1 and NEU1 exerted their roles via Wnt/b-catenin signaling pathway. Overexpression of TOB1-AS1 blocked GC development in vivo. Mechanically, miR-23a was targeted by TOB1-AS1, but directly targeted NEU1. CONCLUSIONS: TOB1-AS1/miR-23a/NEU1 axis regulated proliferation, apoptosis, migration, and invasion of GC cells via Wnt/b-catenin pathway, providing the evidence for serving TOB1-AS1 as an underlying therapeutic target in human GC treatment.

[Role of a liver pathology standardized scoring system in the diagnosis of congenital biliary atresia and its relationship with prognosis].

Objective: To evaluate the diagnostic value of a histologic scoring system in congenital biliary atresia and its prognostic relevance. Methods: From January 2017 to June 2018 at Children's Hospital of Fudan University, 172 wedge liver biopsy specimens were obtained from infants with neonatal cholestasis [119 patients with congenital biliary atresia (CBA) and 53 patients with non-obstructive cholestasis as control]. A pathologist, single-blinded to the final diagnosis, made the histological diagnosis individually based on an 8-feature (portal ductal proliferation, bile duct reaction, bile plugs in portal ductules, liver fibrosis, edema in portal region, cholestasis, inflammatory cells infiltration in portal region, and ductal plate malformation), 21-point scoring system. Results: The main pathologic changes of biliary atresia were hepatocyte cholestasis, hyperplasia of bile ducts, fibrosis and infiltration of inflammatory cells in the portal area. There were significant difference in the degree of portal edema, bile duct hyperplasia and fibrosis between two groups (P<0.01). In addition, there were characteristic bile duct thrombosis in 97.5%(116/119) of the cases and abnormal development of bile duct plate in 9.2%(11/119) of the cases. Compared with non-CBA infant cholestasis group, the difference was statistically significant (P<0.05). The scoring system has high sensitivity, specificity (both 94.1%) and accuracy (94.3%) in the diagnosis of CBA. A score equal to or more than 11 points supported a diagnosis of CBA; whereas a score less than 11 points might suggest cholestasis. The degree of hepatic fibrosis and ductal plate malformation were related to prognosis. Conclusions: The liver pathology scoring system (8-feature, 21-point) is more accurate in diagnosing CBA than previous methods, which may guide the clinicopathological diagnosis. This histological scoring system also helps to assess the prognosis of CBA.

[Clinicopathological analysis of infantile/congenital fibrosarcomas with rare histological features].

Objective: To analyze the clinicopathological features, immunohistochemical (IHC) phentotype,diagnosis and differential diagnosis of infantile/congenital fibrosarcoma (IFS/CFS) with unusual histological features. Methods: Five IFS/CFS at Children's Hospital of Fudan University from March 2014 to July 2018 were analyzed for their diagnosis and differential diagnosis. Results: Two cases were males, three cases were females. The clinical manifestation of IFS/CFS was a rapidly-growing and painless mass. There were no specific radiologic features. Histologically, the tumor cells are arranged in intersecting or sheet-like patterns. There were focal hemangioma-like areas in four cases. There were also focal areas of primitive asteroid, short-spindled, and oval tumor cells in three cases. IHC study showed the tumor cells diffusely expressed TLE1(2/5), Vimentin(5/5), and WT1(3/5), in a cytoplastic pattern; they focally expressed CD34(3/5), CD31(3/5), and α-SMA(2/5). Fluorescence in situ hybridization (FISH) detected break-apart positivity of ETV6 gene. Conclusions: Hemangioma-like pattern, myxoid area, and TLE1 expression is very rare in IFS/CFS. Detection of ETV6 gene break-apart by FISH is very helpful in the diagnosis and differential diagnosis of IFS/CFS.