Fibroblasts overpressing WNT2b cause impairment of intestinal mucosal barrier / 南方医科大学学报

OBJECTIVE@#To investigate the mechanism by which fibroblasts with high WNT2b expression causes intestinal mucosa barrier disruption and promote the progression of inflammatory bowel disease (IBD).@*METHODS@#Caco-2 cells were treated with 20% fibroblast conditioned medium or co-cultured with fibroblasts highly expressing WNT2b, with the cells without treatment with the conditioned medium and cells co-cultured with wild-type fibroblasts as the control groups. The changes in barrier permeability of Caco-2 cells were assessed by measuring transmembrane resistance and Lucifer Yellow permeability. In Caco-2 cells co-cultured with WNT2b-overexpressing or control intestinal fibroblasts, nuclear entry of β-catenin was detected with immunofluorescence assay, and the expressions of tight junction proteins ZO-1 and E-cadherin were detected with Western blotting. In a C57 mouse model of dextran sulfate sodium (DSS)-induced IBD-like enteritis, the therapeutic effect of intraperitoneal injection of salinomycin (5 mg/kg, an inhibitor of WNT/β-catenin signaling pathway) was evaluated by observing the changes in intestinal inflammation and detecting the expressions of tight junction proteins.@*RESULTS@#In the coculture system, WNT2b overexpression in the fibroblasts significantly promoted nuclear entry of β-catenin (P < 0.01) and decreased the expressions of tight junction proteins in Caco-2 cells; knockdown of FZD4 expression in Caco-2 cells obviously reversed this effect. In DSS-treated mice, salinomycin treatment significantly reduced intestinal inflammation and increased the expressions of tight junction proteins in the intestinal mucosa.@*CONCLUSION@#Intestinal fibroblasts overexpressing WNT2b causes impairment of intestinal mucosal barrier function and can be a potential target for treatment of IBD.

Mechanism of intestinal injury induced by WNT2B high-expressed fibroblasts in Crohn's disease / 中华儿科杂志

Objective: To explore the mechanism of intestinal tissue damage induced by macrophages activated by WNT2B high-expressed fibroblasts. Methods: This study involved biological information analysis, pathological tissue research and cell experimental research. The biological information of the colon tissue from the children with inflammatory bowel disease in previous study was analyzed again with single-cell sequencing. The pathological tissues were collected by colonoscopy from 10 children with Crohn's disease treated in the Department of Gastroenterology of Guangzhou Women and Children's Medical Center from July 2022 to September 2022. According to the findings of colonoscopy, tissues with obvious inflammation or ulceration were classified as the inflammatory group, while tissues with slight inflammation and no ulceration were classified as the non-inflammatory group. HE staining was performed to observe the pathological changes of the colon tissues. Macrophage infiltration and CXCL12 expression were detected by immunofluorescence. In terms of cell experiments, fibroblasts transfected with WNT2B plasmid or empty plasmid were co-cultured with salinomycin treated or non-treated macrophages, respectively; the expression of proteins through Wnt classical pathway were detected by western blotting. Macrophages treated with SKL2001 were used as the experimental group, and those with phosphate buffer as the control group. The expression and secretion of CXCL12 in macrophages were detected by quantitative Real-time PCR and enzyme-linked immunosorbent assay (ELISA). T-test or rank sum test were used for the comparison between groups. Results: Single-cell sequencing analysis suggested that macrophages were the main cells in inflammatory bowel disease colon tissue, and there was interaction between WNT2B high-expressed fibroblasts and macrophages. HE staining of the 10 patients ((9.3±3.8) years old, 7 males and 3 females) showed that the pathological score of colon tissue in the inflammatory group was higher than that in the non-inflammatory group (4 (3, 4) vs. 2 (1, 2) points, Z=3.05, P=0.002). Tissue immunofluorescence indicated that the number of infiltrating macrophages in the inflammatory group was significantly higher than that in the non-inflammatory group under high power field of view (72.8±10.4 vs.8.4±3.5, t=25.10, P<0.001), as well as the number of cells expressing CXCL12 (14.0±3.5 vs. 4.7±1.9, t=14.68, P<0.001). In cell experiments, western blotting suggested an elevated level of glycogen synthase kinase-3β phosphorylation in macrophages co-cultured with fibroblast transfected with WNT2B plasmid, and salinmycin could reverse this change. Real-time PCR suggested that the transcription level of CXCL12 in the experimental group was higher than that in the control group (6.42±0.04 vs. 1.00±0.03, t=183.00, P<0.001), as well as the expression and secretion of CXCL12 by ELISA ((465±34) vs. (77±9) ng/L, t=13.21, P=0.006). Conclusion: WNT2B high-expressed fibroblasts can secrete WNT2B protein and activate the Wnt classical signaling pathway thus enhancing the expression and secretion of CXCL12 in macrophages, inducing the development of intestinal inflammation of Crohn's disease.

Meranzin Hydrate Improves Depression-Like Behaviors and Hypomotility via Ghrelin and Neurocircuitry / 中国结合医学杂志

OBJECTIVE@#To investigate whether meranzin hydrate (MH) can alleviate depression-like behavior and hypomotility similar to Chaihu Shugan Powder (CSP), and further explore the potential common mechanisms.@*METHODS@#Totally 120 Spraque-Dawley rats were randomly divided into 5-8 groups including sham, vehicle, fluoxetine (20 mg/kg), mosapride (10 mg/kg), CSP (30 g/kg), MH (9.18 mg/kg), [D-Lys3]-GHRP-6 (Dlys, 0.5 mg/kg), and MH+Dlys groups by a random number table, 8 rats in each group. And 32 mice were randomly divided into wild-type, MH (18 mg/kg), growth hormone secretagogue receptor-knockout (GHSR-KO), and GHSR+MH groups, 8 mice in each group. The forced swimming test (FST), open field test (OFT), tail suspension test (TST), gastric emptying (GE) test, and intestinal transit (IT) test were used to assess antidepressant and prokinetic (AP) effects after drug single administration for 30 min with absorbable identification in rats and mice, respectively. The protein expression levels of brain-derived neurotrophic factor (BDNF) and phosphorylated mammalian target of rapamycin (p-mTOR) in the hippocampus of rats were evaluated by Western blot. The differences in functional brain changes were determined via 7.0 T functional magnetic resonance imaging-blood oxygen level-dependent (fMRI-BOLD).@*RESULTS@#MH treatment improved depression-like behavior (FST, OFT) and hypomotility (GE, IT) in the acute forced swimming (FS) rats (all P<0.05), and the effects are similar to the parent formula CSP. The ghrelin antagonist [D-Lys3]-GHRP-6 inhibited the effect of MH on FST and GE (P<0.05). Similarly, MH treatment also alleviated depression-like behavior (FST, TST) in the wild-type mice, however, no effects were found in the GHSR KO mice. Additionally, administration of MH significantly stimulated BDNF and p-mTOR protein expressions in the hippocampus (both P<0.01), which were also prevented by [D-Lys3]-GHRP-6 (P<0.01). Besides, 3 main BOLD foci following acute FS rats implicated activity in hippocampus-thalamus-basal ganglia (HTB) circuits. The [D-Lys3]-GHRP-6 synchronously inhibited BOLD HTB foci. As expected, prokinetic mosapride only had effects on the thalamus and basal ganglia, but not on the hippocampus. Within the HTB, the hippocampus is implicated in depression and FD.@*CONCLUSIONS@#MH accounts for part of AP effects of parent formula CSP in acute FS rats, mainly via ghrelin-related shared regulation coupled to BOLD signals in brain areas. This novel functionally connection of HTB following acute stress, treatment, and regulation highlights anti-depression unified theory.

Analysis of Radial Peripapillary Capillary Density in Patients with Bietti Crystalline Dystrophy by Optical Coherence Tomography Angiography / 生物医学与环境科学（英文）

OBJECTIVE@#We wanted to investigate the radial peripapillary capillary (RPC) network in patients with Bietti crystalline dystrophy (BCD).@*METHODS@#We compared RPC densities in the disk and different peripapillary regions, obtained using optical coherence tomography angiography in 22 patients with BCD (37 eyes) and 22 healthy subjects (37 eyes). The BCD group was then divided into Stage 2 and Stage 3 subgroups based on Yuzawa staging, comparing the RPC densities of the two.@*RESULTS@#The disk area RPC density was 38.8% ± 6.3% in the BCD group and 49.2% ± 6.1% in the control group ( P < 0.001), and peripapillary region RPC density was significantly lower in the BCD group than in the control group (49.1% ± 4.7% and 54.1% ± 3.0%, respectively, P < 0.001). There were no significant RPC density differences between the tempo quadrant and inside disk of Stages 2 and 3 subgroups; the other areas showed a significantly lower RPC density in Stage 3 than in Stage 2 BCD.@*CONCLUSION@#The BCD group RPC density was significantly lower than the control group. The reduction of RPC density in the tempo quadrant occurred mainly in the Stage 1 BCD. In contrast, the reduction of RPC density in superior, inferior, and nasal quadrants occurred mainly in Stage 2.

The predictive value and efficacy of the risk model based on the metabolic parameters of 18F-FDG PET-CT for EGFR gene mutations in non-small cell lung cancer / 中华放射学杂志

Objective:To explore the value and efficacy of the risk model based on the metabolic parameters of 18F-FDG PET-CT in predicting epidermal growth factor receptor (EGFR) gene mutations in non-small cell lung cancer (NSCLC). Methods:This retrospectives study reviewed 105 NSCLC patients who were tested for EGFR gene expression and underwent 18F-FDG PET-CT exam prior to treatment from Jan 2017 to June 2018 in our hospital. The patients were divided into EGFR mutations group ( n=40) and EGFR wild type group ( n=65). The differences between the different groups were analyzed in several clinical characteristics and three metabolic parameters based on 18F-FDG PET-CT, including the maximal standard uptake value (SUV max), metabolic tumor volume (MTV), total lesion glycolysis (TLG) of the primary tumor. Multivariate logistic regression analysis was performed to identify predictors of EGFR mutations, and the risk prediction model and nomogram graph were constructed. Diagnostic efficiency of the model was done by the receiver operating characteristics (ROC) curve analysis, and the Calibration plot was performed by Hosmer-Lemeshow (H-L) test to evaluate the calibration scale of the model. Results:There were statistically significant differences in gender, smoking status, serum CEA level, length of tumor, pathological types, TTF-1 and NapsinA expression between the EGFR mutant groups and EGFR wild-type groups (all P<0.05). The MTV and TLG of EGFR mutation group were 4.4 (4.5，37.1) cm 3 and 46.6 (21.2，118.2), respectively. The MTV and TLG of EGFR wild type group were 7.4 (3.2，13.5) cm 3 and 95.4 (26.4，345.1), respectively. The MTV and TLG of EGFR mutation group were significantly lower than those of EGFR wild type group ( Z=-2.452, P=0.014; Z=-2.379, P=0.017). ROC curve analysis showed area under the curve (AUC) predicted by SUV max, MTV and TLG for EGFR mutations was 0.597, 0.643 and 0.639, respectively. Multivariate analysis demonstrated that gender, length of tumor, SUV max and MTV were independent predictors of EGFR mutations, with the odds ratio (OR) values (95 %CI) as 3.811 (1.508-9.629), 1.679 (0.899-3.136), 0.928 (0.848-1.015) and 0.924 (0.865-0.986), respectively. The predictive model and nomogram graph was established, with the sensitivity, specificity, positive predictive value, negative predictive value and AUC of 80.0%, 66.2%, 68.8%, 75.3% and 0.775 (0.687-0.864), respectively. The H-L test showed the model had excellent accuracy (χ 2=3.872, P=0.869). Conclusion:The risk model based on the metabolic parameters of 18F-FDG PET-CT has a good performance in predicting the mutations of EGFR gene in patients with NSCLC.

Benign metastasizing leiomyoma: A case report andliterature review / 中南大学学报(医学版)

Benign metastasizing leiomyoma (BML) often occurs in women with a history of uterine leiomyoma at the childbearing age. At the same time, BML cases with multiple sites of metastatic lesions are extremely rare. A BML patient with multiple metastases of uterine leiomyoma in two lung and lumbar spine after surgery was admitted to Department of Spinal Surgery, Second Xiangya Hospital, Central South University in September 2017. Positron emission tomography/computed tomography (PET/CT) can perform a whole body examination for BML patients, which can find metastases in many parts of the body such as lung, abdominal cavity, pelvic cavity and spine. The PET/CT results are helpful to a comprehensive diagnosis. The imaging and clinical features of BML are now explored in combination with the characteristics of the case and relevant literature reports.

18F-FDG PET/CT features on paraganglioma of testis with multiple lymph node and lung metastasis: A case report and literature review / 中南大学学报(医学版)

To determine the clinicopathological and imaging features in 18F-fluoro-2-deoxyglucose (18F-FDG) positron emission tomography and computed tomography (PET/CT) for paraganglioma of testis, and to increase the diagnostic accuracy.
 Methods: A case of paraganglioma of testis with multiple lymph node and lung metastasis were reported. PET/CT and pathological findings in the case were retrospectively analyzed.
 Results: The patient presented with high blood pressure, high level of catecholamine, and urinary vanillylmandelic acid. The patient underwent 18F-FDG PET/CT, which showed the features including the right testis nodule with a star lesion nearby, the right spermatic cord, the lymphadenopathy of bilateral inguinal and retroperitoneum, the posterior basal segment of right lung nodule, and a lot of brown adipose tissues (BAT) in the whole body with intense FDG uptake. 18F-FDG PET/CT showed that the intense FDG uptake of the BAT disappeared after the excision of the right testis and metastasis of paraganglioma.
 Conclusion: PET/CT shows great value in localization diagnose, clinical staging and curative evaluation. PET/CT plays a helpful role in revealing the BAT with 18F-DG avidity in the patients with paraganglioma with elevated blood pressure, high level catecholamine, and urinary vanillylmandelic acid.

Comparative research of setup error and registration for image-guided radiotherapy of lung cancer / 医疗卫生装备

Objective To explore the setup error and area registration error during lung cancer radiotherapy by using the on board imager (OBI) of the linear accelerator. Methods Totally 50 lung cancer patients underwent image-guided radiation therapy. Then OBI system was used for the scan validation by electronic portal imaging device (EPID) and cone beam CT (CBCT), and comparative analysis was executed on the setup errors of EPID and CBCT. Results The translation errors of EPID were (-1.62 ±1.58), (2.12 ±1.49) and (4.52 ±2.42)mm respectively at Lat, Vrt and Lng directions, while those of CBCT were (-1.27±1.25), (1.43±1.57) and (3.12±2.62) mm respectively. The registration errors at Lat, Vrt and Lng directions and rotation angle of lung tissue were (-1.27±1.25), (1.43±1.57), (3.12±2.62)mm and (0.5±1.6)° respectively, and those of target area were (-1.56±1.78), (1.68±2.39), (3.42±2.73)mm and (0.8±1.9)° respectively. CBCT and EPID had statistical differences (P<0.05) in setup error validation as well as setup errors at Vrt and Lng directions. There were no significant differences (P>0.05) when CBCT self-registration was involved in selecting different areas. Conclusion CBCT and EPID can both used for the setup validation of lung cancer, while the former behaved better than the latter.

Surgical Treatment for Myelomeningocele Associated With Hydrocephalus in Neonate(6 Case Report and Review of the Literature) / 中国医师杂志

Objective To determine pathogenesis and the suitable time of operation for myelomeningocele associated with hydrocephalus in neonate.Methods 6 underwent head CT scanning, 2 lumbosacral CT scanning and 6 lumbosacral X radiography on 6 patients myelomeningocele complicated with hydrocephalus.Ventriculoperitoneal shunt and repair of the myelomeningocele were performed respectively for one patient.from 1 day to 28 day old.Operation stage 1 in 5 patients.Repair of the myelomeningocele concurred with ventriculoperitoneal shunt. Intracranial pressure was measured in shunting procedure.Results 4 patients had normal intracranial pressure,2 patients increased intracranial pressure in the 6 patients.The volume of the hernial sac had markedly diminished and status of hernial sac had greatly improved wall in the patient who wnderwent two-stage procedures after shunt procedure. Lumbosacral wound healing was good . No recurrent myelomeningocele was found, no hydrocephalus was seen using head CT scanning and clinical manifestation has improved in these patients who were followed up 6 month to 3 year.Conclusions Hydrocephalus may deteriorate the degree of lumbosacral myelomeningocele. Effecacy of vntriculoperitoneal shunt and repair of the myelomeningocele was excellent in myelomeningocele complicated with hydrocephalus in neonate.Micro-operative technique might prevent the occurrence of tethered cord.