[Study on in vitro differentiation of human adenoid-derived mesenchymal stem cells into olfactory sensory neurons].

Objective: To investigate the feasibility of isolation and culture of human adenoid-derived mesenchymal stem cells (aMSCs) in vitro, and to observe the differentiation of aMSCs into olfactory sensory neurons. Methods: Adenoid tissues surgically removed from children with adenoid hypertrophy in the Second Xiangya Hospital of Central South University from September to November of 2020 were collected. The adenoid tissues were digested and isolated by trypsin and then cultured with adhesion method. The expressions of cell surface antigens CD45, CD73 and CD90 on aMSCs of P5 generation were tested by flow cytometry, and the ability of osteogenic and adipogenic induction were used to identify cell differentiation ability. Then, aMSCs were induced into differentiation by retinoic acid (RA), sonic hedgehog (SHH), basic fibroblast growth factor (bFGF), RA+SHH, RA+bFGF, SHH+bFGF and RA+SHH+bFGF, respectively. The morphology of differentiated cells was observed under inverted microscope. The expression of ß-tubulin 3, which was the specific marker of sensory neuron, the expressions of growth associated protein-43 (GAP43) and olfactory maker protein (OMP), which were the specific markers of olfactory sensory neuron, were detected by immunofluorescence antibody assay. The expression intensities were compared by Chi-square test of four-grid table data. Results: aMSCs were successively isolated and cultured from human adenoid tissues. P0 cells generation had good adhesion and proliferation performance. P2 cells were basically purified. P5 cells expressed CD73 and CD90 with the purity of 99.3% and 99.75% respectively, without CD45 expression. P5 cells had a good ability of osteogenic differentiation and adipogenic differentiation. Neuron-like morphology and expression of ß-tubulin 3 were found in differentiated cells after induced by RA, SHH, or bFGF, respectively. An induction of expression of GAP43 was found in differentiated cells of bFGF+SHH group and RA+SHH+bFGF group, without expression of OMP of each group. The intensity of GAP43 expression of RA+SHH+bFGF group was stronger than that of bFGF+SHH group (χ2=17.48, P<0.005). Conclusions: aMSCs can be cultured from human adenoid tissues, with the stably passaged and good differentiation ability. As a new population of mesenchymal stem cells, aMSCs have the neuroregenerative properties and could differentiate into immature olfactory sensory neurons under the induction of RA+SHH+bFGF in vitro.

[Role of up-regulated DDX3 in the proliferation of human cervical cancer cells].

Objective: To investigate the role of DDX3 up-regulation in the proliferation of human cervical cancer cells and its correlation with clinical prognosis. Methods: Expression levels of DDX3 in the 59 specimens of cervical cancer and adjacent non-neoplastic tissue collected at Henan Provincial People's Hospital from April 2012 to March 2013 were detected using immunohistochemistry. A lentivirus-mediated DDX3-over-expression cell line was constructed based on HeLa cells of cervical cancer. CCK-8 assay was used to evaluate cell survival rate. Boyden chamber was used to measure the cell migration and invasion. Real-time fluorescence quantitative PCR was used to detect DDX3 expression level and Western blot was used to detect the expression of EMT and PI3K/Akt signal pathway-related proteins. Results: DDX3 overexpression was associated with FIGO stage, depth of cervical invasion and lymph node metastasis (P<0.05). Kaplan-Meier analysis revealed that cervical cancer patients with high expression of DDX3 had a poor overall survival (P<0.05). Compared with the cells transfected with pLVX-Con vector, the expression of DDX3 protein and mRNA was significantly increased in the cells transfected with pLVX-DDX3 (all P<0.01). Cell proliferation was significantly increased following transfection with pLVX-DDX3 for 72 h in HeLa cells compared with that transfected with pLVX-Con (P<0.05). Compared with the controls, DDX3 overexpression significantly promoted the migration and invasion of HeLa cells (P<0.05), and increased the expression of N-Cadherin, vimentin and Snail in HeLa cells (P<0.05). In pLVX-DDX3 group, the expression levels of ß-catenin, phosphorylated Akt, and pAkt's downstream target p-GSK3ß were significantly higher than those of pLVX-Con group (P<0.05). The expression levels of p-Akt, p-GSK3ß and ß-catenin were decreased when the PI3K/Akt pathway was blocked using the PI3K inhibitor LY294002 (P<0.05), and the expression levels of N-Cadherin, vimentin and Snail were also significantly decreased (P<0.05). Conclusions: DDX3 overexpression promotes proliferation, migration and invasion of cervical cancer cells, and induces epithelial-mesenchymal transition (EMT). Its mechanism may be related to activation of the PI3K/Akt signaling pathway.

[Study on the relationship between lifestyle and depression symptoms: a TCLSIH study].

Objective: To evaluate the relationship between lifestyle factors and depressive symptoms based on the TCLSIH cohort of 2013-2016 and provide evidence for the intervention on lifestyle in the prevention and treatment of depression in the future. Methods: A cross-sectional study was conducted in 24 256 persons by using a self-rating depression scale (SDS) to assess the depressive symptoms, and lifestyle questionnaire survey and physical examination were carried out. By using software SAS 9.3. The study subjects were divided into two groups: non-depression group (SDS score <45) and depression group (SDS score≥45), and the relationship between lifestyle factors and depressive symptoms was analyzed. Results: The study subjects in depression group accounted for 16.59%; the baseline survey showed that compared with non-depression group, the subjects in depression group had higher neutrophil count and lymphocyte count ratio (NLR), lower BMI, lower total energy intake, and lower physical activity level, and tended to take less plant food diet, more animal food diet and sweet food diet (P<0.000 1). In the depressive group, there were more smokers and less occasional drinkers (P<0.01), and there were more women, home-aloners, people with lower education levels, people with lower total household income, and less staff members, married and those who liked to contact relatives and friends, but the proportion of people who spent more than 5 hours daily for outdoor activities was higher. Multiple linear regression analysis results showed that being male, aged, NLR, smoking (OR=1.14, 95%CI: 1.03-1.26), quitting alcohol, being home-aloners, animal food diet (OR=1.41, 95%CI:1.35-1.46), sweet food diet (OR=1.17, 95%CI: 1.13-1.22), sleep time >7.5 h/d, outdoor activity time 3-5 h/d, outdoor activity time >5 h/d were positively correlated with depression (P<0.05). BMI(OR=0.98, 95%CI: 0.97-0.99), education level (OR=0.76, 95%CI: 0.70-0.82), being staff member, total household income (OR=0.63, 95%CI: 0.58-0.68), total energy intake, physical activity (OR=0.86, 95%CI: 0.84-0.89), married status, move contacts with relatives or friends were negatively related with depression (P<0.05). Conclusion: Lifestyle is closely related to the occurrence of depressive symptoms, and lifestyle intervention seems be a new way to prevent and treat depression.

Long non-coding RNA OR3A4 promotes metastasis of ovarian cancer via inhibiting KLF6.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Long non-coding RNA OR3A4 promotes metastasis of ovarian cancer via inhibiting KLF6, by F.-F. Guo, M.-M. Jiang, L.-L. Hong, B. Qiao, X.-M. Lin, W.-Y. Xu, X.-Q. Fu, published in Eur Rev Med Pharmacol Sci 2019; 23 (6): 2360-2365-DOI: 10.26355/eurrev_201903_17380-PMID: 30964160" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/17380.

[Transjugular intrahepatic portosystemic shunt for the treatment cavernous transformation of the portal vein with vareceal bleeding].

Objective: To evaluate the efficacy and safety of transjugular intrahepatic portosystemic shunt(TIPS) for the treatment of patients with cavernous transformation of portal vein (CTPV) with vareceal bleeding. Methods: From September 2016 to June 2018, a total of 21 patients suffered CTPV complicated with vareceal bleeding were admitted to First Affiliated Hospital of Zhengzhou University. TIPS were performed combined with percutaneous transhepatic portal vein assist. There were 13 males and 8 females, with an average age of 27-67 (48±11) years. Blood routine examination, liver function test, blood ammonia and ultrasound Doppler were conducted 1,3,6 months after operation, and every 6 months during follow-up. Abdominal enhanced CT and digital substraction angiography were followed every year. Results: TIPS were successfully performed in 19 cases (90.5%), esophageal and gastric varices were embolized in 17 cases; 2 cases failed to selective catheterized, then endoscopic therapy was performed.All bleeding stopped after operation. The pressure of portal vein decreased from 25.0-44.0 (33.7±5.4) mmHg (1 mmHg=0.133 kPa) to 17.0-30.0 (24.5±3.1) mmHg, portosystemic pressure gradient decreased from 16.0-32.0 (23.5±4.6) mmHg to 9.0-15.0 (11.4±1.9) mmHg after TIPS (all P<0.05). During 3-24 months follow-up, 2 patients suffered from hepatic encephalopathy, 3 patients had recurrent upper gastrointestinal bleeding, including 1 duodenal ulcer and 2 esophageal varices. In-stent restenosis were found in 6 patients,in which 3 patients underwent shunt revision operation. At the end of the follow-up, the cumulative patency was 16/19. Conclusion: For patients with CTPV and vareceal bleeding, TIPS could reduce portal hypertension while embolizing varicose veins.It is a safe and effective treatment.

Long non-coding RNA OR3A4 promotes metastasis of ovarian cancer via inhibiting KLF6.

OBJECTIVE: Recently, long noncoding RNAs (lncRNAs) have attracted much attention for their roles in tumor progression. The aim of this study was to investigate the exact role of lncRNA OR3A4 in the development of ovarian cancer (OC), and to explore the possible underlying mechanism. PATIENTS AND METHODS: OR3A4 expression in OC tissue samples was detected by Real-time quantitative polymerase chain reaction (qRT-PCR). Moreover, wound healing assay and transwell assay were performed to evaluate the effect of OR3A4 on the metastasis of OC. Furthermore, the underlying mechanism was explored by RT-qPCR and Western blot assay. RESULTS: The expression level of OR3A4 in OC samples was significantly higher than that of adjacent tissues. Moreover, cell migration and invasion were significantly repressed after OR3A4 knockdown in vitro. Moreover, the mRNA and protein expressions of kruppel-like factor 6 (KLF6) were remarkably upregulated after knockdown of OR3A4. Furthermore, the expression level of KLF6 was negatively correlated with the expression of OR3A4 in OC tissues. CONCLUSIONS: Our results showed that OR3A4 could enhance OC cell metastasis and invasion via suppressing KLF6. Moreover, OR3A4 might be a potential therapeutic target for OC.

[Mutation analysis and early pregnancy prenatal diagnosis for two families affected with non-syndromic hearing loss].

Objective:The aim of this study is to identify the pathogenic genes of two non-syndromic hearing loss families, provide accurate genetic counseling and early pregnancy prenatal diagnosis services for second birth families, which determine the genotype of the fetus, early diagnosis and early intervention to prevent the birth of deaf children. Method:Two families with a severe sensorineural hearing loss were enrolled in this study. Genomic DNA was extracted from the peripheral venous blood of the proband and parents. The mutations of the four hearing loss susceptibility genes were analyzed by the hereditary hearing loss gene detection kit (PCR reverse hybridization) and Sanger sequencing. After confirming the genotype of the subject, the genotype of the fetus was examined for the fetus with 10 to 12 weeks of gestation, and the neonates were diagnosed with prenatal diagnosis. Result:In the two families, the No. 1 family proband was the compound heterozygous mutation of SLC26A4 gene c.IVS72A＞G/c.2177insCTAT, and parents were carriers. The prenatal diagnosis showed that the fetal genotype of No. 1 family was a compound heterozygous mutation, and the fetus was not born. The No. 2 family proband genotype was the compound heterozygous of GJB2 gene c.605ins46/c.512insAACG, and the parents were both carriers. The No. 2 family, the prenatal diagnosis showed the fetal genotype of No. 2 family GJB2 gene sequence was normal, the newborn passed the hearing screening. Conclusion:Genetic testing of hereditary hearing loss combined with prenatal diagnosis plays a significant role in guiding deaf children with re-fertility needs, especially prenatal diagnosis in early pregnancy, which can achieve early diagnosis, early detection and early intervention, effectively reducing the birth rate of children with deafness.

[Expression of type 1 and type 2 cytokines from serum of coal miners and the evaluation in surveillance of coal workers' pneumoconiosis at earlier stage].

Objectives: To explore the expression regulation of type 1 and type 2 (Th1 and Th2) cytokines from serum of coal miners and the evaluation in surveillance of coal workers' pneumoconiosis, 630 coal miners were studied. Methods: A total of 90 male patients diagnosed as coal workers' pneumoconiosis (CWP) in a institute for occupational health and 19 male workers newly diagnosed as CWP patients was chosen as CWP group with simple random sampling method from a coal mine group from January 2013 to December in 2015. 180 male coal miners with abnormal but not diagnosed as CWP were selected as CWP suspected group with simple random sampling methods, meanwhile 180 male coal miners with normal chest X-ray photograph was as dust-exposed group by 1∶1 matched as age. And 161 healthy males accepted pre-employed examination were selected as control group, CWP suspected group, dust-exposed group and control group called as non-CWP group. According to screening test and diagnosis test, the basic information and occupational history of all subjects were collected, and cytokines including IL-1ß, IL-8, IFN-γ, IL-6 and IL-10 of serum were detected. Receiver operator characteristic (ROC) curve was used to determine the optimal cutoff value of each cytokine. Area under curve (AUC), the validity and reliability were calculated and judged. Results: The average age of control group, dust-exposed group, CWP suspected group and CWP group were (27.4±5.0) , (43.4±10.7) , (48.2±6.2) , (64.7±7.0) years old, respectively. The median level of IL-1ß, IL-8, IFN-γ and IL-6 in cases group (1 638.30, 2 099.49, 815.18,140.32 pg/ml) were higher than that of non-cases group (1 445.57, 1 402.26, 736.38, 95.73 pg/ml) (P<0.05) . The level of IL-8 (1 503.99 pg/ml) in CWP suspected group was higher than that of control group (1 295.67 pg/ml) and dust-exposed group (1 376.94 pg/ml) , but the level of IL-10 (654.08 pg/ml) was lower than that of control group (596.64 pg/ml) . The ratio of IFN-γ/IL-6 ranged from 5 to 8, and the ratio in CWP group (5.87) was lower than that of non-CWP group (7.61) . The IL-6 and IL-8 among the subjects of dust-exposed group in terms of the age distribution of among had reached statistical significance. According to ROC, the cutoff value of IL-1ß, IL-6, IL-8, IL-10 and INF-γ reached 1 582.65, 116.53, 1 791.54, 581.08 and 792.69 pg/ml, respectively. The AUC was 0.668, 0.895, 0.859, 0.716 and 0.637, respectively. It was found that IL-6 and IL-8 could be used as biomarkers in detecting CWP, the sensitivity and specificity was 82.6% and 84.6%, 78.0% and 84.8%, respectively; Youden's index was 0.674 and 0.628 and the consistency rate was 84.3% and 83.7%, while Kappa value was 0.55 and 0.52. Conclusion: There was Type 1 and type 2 cytokine dysregulation in CWP patients. IL-6 and IL-8 can be used as effective biomarkers to forecast lung injury before X-ray changes.

MiR-1-3p inhibits cell proliferation and invasion by regulating BDNF-TrkB signaling pathway in bladder cancer.

Recent studies have confirmed the existence of BDNF and tropomyosin-related kinase B (TrkB) in normal and cancerous urothelium. However, the corresponding mechanisms and upstream signal pathways of BDNF/TrkB have not been fully discovered. This study aimed to investigate the effects of miR-1-3p on bladder cancer (BC) by regulating BDNF-TrkB signal pathway. The expression of miR-1-3p and BDNF in BC tissues and cell lines were detected by Cancer Genome Atlas (TCGA) microarray analysis, RT-qPCR and western blot. Cell transfection was done using Lipofectamine 2000. Then cell viability, proliferation, migration and apoptosis were measured by MTT, colony formation assay, Transwell assay and flow cytometry, respectively. The relationship between miR-1-3p and BDNF was confirmed by luciferase reporter gene assay. MiR-1-3p was significantly down-regulated in BC tissues and cell lines, while BDNF was significantly up-regulated compared to normal samples. MiR-1-3p targeted BDNF and suppressed its expression. Transfections of miR-1-3p mimics and BDNF siRNAs can suppress BC cell proliferation, invasion and induce cell apoptosis. In addition, miR-1-3p can inhibit phosphorylation of the TrkB by regulating BDNF.In conclusion, MiR-1-3p has significant effects on viability, proliferation, invasion and apoptosis of BC cells by regulating BDNF-TrkB pathway.

Involvement of ghrelin in nucleus tractus solitaries on gastric signal afferent and gastric motility in cisplatin-treated rats.

OBJECTIVE: Ghrelin had been known to promote gastric motility in human and animals previously. We aim to investigate the role of ghrelin in chemotherapy-induced nausea and vomiting. MATERIALS AND METHODS: In the present study, we observed the changes in food intake, kaolin consumption, body weight, plasma ghrelin concentration and expression of ghrelin and its receptor GHS-R1a in the stomach and nucleus tractus solitaries (NTS) in cisplatin-treated rats, and the effects of ghrelin microinjected into NTS on the discharge activity of gastric distension (GD) responsive neurons and gastric motility were also observed. RESULTS: Cisplatin induced the decrease in food intake and the increase in kaolin consumption of rats. In addition, mRNA expression of GHS-R1a in the stomach and NTS increased significantly after cisplatin treatment. The discharge activity of GD excited (GD-E) and GD inhibited (GD-I) neurons in cisplatin-treated rats was weaker than that of saline treatment, while ghrelin administration into NTS excited most of GD-E and GD-I neurons. Cisplatin induced the decrease in gastric contraction while ghrelin administrated into NTS promoted the gastric motility significantly. However, the amplitude and frequency of gastric contraction promoted by ghrelin in NTS of cisplatin-treated rats were lower than that of saline treated rats. The effects of ghrelin could be completely blocked by its receptor antagonist BIM28163. CONCLUSIONS: These results indicated that ghrelin in the NTS might participate in the regulation of GD-neurons and gastric motility via its receptor in cisplatin-treated rats.

Successful use of the dilated right spermatic cord vein for renal transplantation in a patient with congenital hypoplasia of the inferior vena cava.

A 29-year-old man was admitted to our department with renal failure secondary to glomerulonephritis. No history of deep venous thromboses was reported, and no iliac vessel abnormality was evident on routine ultrasound (B-mode) examination before the operation. Transplantation of his mother's left kidney revealed occlusion of his common iliac vein and distal inferior vena cava (IVC). The right spermatic cord vein was noted to be dilated and suitable for venous drainage of the allograft, which was accomplished by an end-to-side anastomosis between the renal vein and the right spermatic cord vein. The allograft showed immediate function; serum creatinine was decreased to a normal value at 5 days after surgery. After the operation, a vascular spiral computerized tomographic 3-dimensional reconstruction showed absence of the infrarenal IVC with the right spermatic cord vein draining into the end of IVC. Physical examination revealed a right-side varicocele with dilated epigastric vein. The donor kidney slower normal values upon routine follow-up at 2 years after the operation.

Clinical analysis of living related renal transplantation with donors older than 50 years in China.

OBJECTIVE: The objective of this study was to investigate whether kidney grafts from living related donors older than 50 years were safe for the donors and recipients in the long term. METHODS: One hundred seven living related donor kidney transplantations were performed in our center from April 1994 to December 2007. No prisoners or organs from prisoners were used in the collection of these data. Donors were divided into 2 groups: >50 years of age (range, 51-78 years), designated as the study group, and ≤50 years of age (range, 21-50 years), designated as the control groups. The mean time of follow-up was 49 months (range, 12-180 months). Clinical data were compared, including donor serum creatinine (Scr) levels, glomerular filtration rates (GFR) before and after the procedures operative complications, and postoperative short-term and long-term recovery of renal function in recipients as well as their complications and recipient and kidney survivals. RESULTS: All operations were successfully performed. Before the operation, the mean Scr and GFR were 82.16 ± 10.86 umol/L and 85.82 ± 6.26 mL/min, respectively, in the study group versus 78.66 ± 10.41 umol/L and 88.74 ± 9.44 mL/min, respectively, in the control group. There were no significant differences in mean Scr or GFR values between the groups at various preoperative or postoperative times (P > .05). No severe perioperative complications occurred, and no subsequent renal function failure was observed upon long-term follow-up of donors in the 2 groups. Comparisons of recipient age, gender ratio, duration on dialysis, HLA matches, cold/warm ischemia times, and immunosuppression therapy showed a correlations between the 2 groups. Mean Scr levels of recipients, which were compared from 1 week to 3 years following surgery, were slightly higher among the control than the study group, but the difference was not significant (P > .05). There were no significant differences between the study and control groups in 1-,3-,5-, and 8-year recipient/graft survival rates (P > .05). CONCLUSIONS: Long-term follow-up showed that transplantations using grafts from donors older than 50 years of age yielded similar results to those with younger donors.

A CalDAG-GEFI/Rap1/B-Raf cassette couples M(1) muscarinic acetylcholine receptors to the activation of ERK1/2.

In this study we examine signaling pathways linking the M(1) subtype of muscarinic acetylcholine receptor (M(1) mAChR) to activation of extracellular signal-regulated kinases (ERK) 1 and 2 in neuronal PC12D cells. We first show that activation of ERK1/2 by the M(1) mAChR agonist carbachol takes place primarily via a Ras-independent pathway that depends largely upon Rap1, another small GTP-binding protein in the Ras family. Rap1 in turn activates B-Raf, an upstream activator of ERK1/2. Consistent with these results, carbachol was found to activate Rap1 more potently than Ras. Similar to other small GTP-binding proteins, activation of Rap1 requires a guanine nucleotide exchange factor (GEF) to promote its conversion from the GDP- to GTP-bound form. Using specific antibodies, we show that a recently identified Rap1 GEF, calcium- and diacylglycerol-regulated guanine nucleotide exchange factor I (CalDAG-GEFI), is expressed in PC12D cells and that carbachol stimulates the formation of a complex containing CalDAG-GEFI, Rap1, and activated B-Raf. Finally, we show that expression of CalDAG-GEFI antisense RNA largely blocks carbachol-stimulated activation of hemagglutinin (HA)1-tagged B-Raf and formation of the CalDAG-GEFI/Rap1/HA1-tagged B-Raf complex. Together, these data define a novel signaling pathway for M(1) mAChR, where increases in Ca(2+) and diacylglycerol stimulate the sequential activation of CalDAG-GEFI, Rap1, and B-Raf, resulting in the activation of MEK and ERK1/2.