[Analysis and significance of HBV DNA below the lower detection limit of HBV RNA levels after long-term NAs antiviral therapy in patients with hepatitis B virus cirrhosis].

Objective: To analyze the significance of HBV DNA below the lower detection limit of HBV RNA levels after long-term nucleos(t)ide analogues (NAs) antiviral therapy in patients with hepatitis B virus cirrhosis. Methods: 97 cases with hepatitis B virus cirrhosis treated with NAs antiviral therapy for at least 3 years between May 2018 to July 2019 were selected. High-sensitivity HBV DNA (<20 IU/ml), alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyltransferase (GGT), HBsAg, HBeAg and HBV RNA at least twice every 6 months were detected. According to Child-Pugh classification, HBeAg, HBsAg level, and HBV RNA level intergroup comparison was performed. Rank sum test, χ2 test and linear regression analysis were performed on the data. Results: Compared with the HBV RNA level of child-Pugh class A patients, the HBV RNA level of Child-Pugh class B+C patients were significantly higher [4.1 (0,4.9) log10 copies/ml and 2.0 (0,3.5) log10 copies/ml], and the difference was statistically significant (Z=2.370, P<0.05). According to different HBeAg levels, they were divided into HBeAg positive and negative group, and the quantitative comparison of HBV RNA levels between the two groups were 2.0 (0, 4.5) log10 copies/ml and 1.0 (1.0, 2.0) log10 copies/ml, respectively, and the difference was statistically significant (Z=3.233, P<0.05). According to different HBsAg levels, they were divided into three groups: HBsAg≤100 IU/ml, 100

[The clinical observation of sirolimus combined with calcineurin inhibitors for steroid-resistant/steroid-dependent extensive cGVHD].

Objective: To observe the efficacy and safety of sirolimus combined with calcineurin inhibitor (CNI) in the treatment of glucocorticoid resistant/dependent extensive chronic graft-versus-host disease (cGVHD) . Methods: A total of 27 patients with steroid-resistant/steroid-dependent extensive cGVHD from November 2015 to January 2019 were enrolled and given sirolimus capsules combined with cyclosporine or tacrolimus to observe the clinical efficacy and adverse events. Results: The median duration of medication was 14.2 months and the mean duration was 16.7 months. The median follow-up time was 20.1 months (12.9-46.1 months) . Following the 6-month follow-up, 3 cases achieved complete response (CR) and 12 cases partial response (PR) . The overall response rate (ORR) was 55.6% ; for progression-free survival (PFS) , PFS-6 reached 88.9% (24/27) , and for overall survival (OS) , OS-6 was 100% . At the 1-year follow-up, there were 5 cases of CR and 11 cases of PR, ORR was 59.3% , PFS-12 reached 62.9% (17/27) , and OS-12 was 100% . The subgroup analysis found that the program was more effective for cGVHD in male donors and the target organ analysis had an advantage in the treatment of oral cavity, skin, and liver rejection. Adverse events were observed: hyperlipidemia 11.1% , oral ulcer 7.4% , fungal infection 11.1% , liver injury 3.7% , renal insufficiency 0, and no new CMV and EB viremia. Conclusion: Sirolimus combined with calcineurin inhibitors is effective in treating steroid-resistant/steroid-dependent extensive cGVHD, especially because adverse reactions (renal toxicity, CMV, EBV infection) are low in number, which is suitable for long-term treatment of cGVHD.

[The clinical observation of serum specific biomarkers in patients with chronic graft-versus-host disease].

Objective: Chronic graft-versus-host disease (cGVHD) is a major long-term complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT) . It is important to study the changes of serum biomarkers expression in patients for early diagnosis and treatment. Methods: The expression levels of five serum protein markers (IL-1b, IL-16, CXCL9, CCL19, CCL17) in patients with or without cGVHD after allo-HSCT were detected by liquid suspension microarray. Results: Compared with the control group without cGVHD, the expression levels of CXCL9 and CCL17 in serum of patients with cGVHD were significantly increased (P<0.05) . CCL17 was correlated with the severity of cGVHD (P<0.001) . CXCL9 was significantly increased in the serum of patients with skin lesion (P<0.01) , and CCL17 was significantly expressed in cGVHD patients with liver as the target organ (P<0.01) . Conclusion: The combination of CXCL9 and CCL17 can be used as serum biomarkers of cGVHD, which has certain reference value in assisting the diagnosis and evaluation of cGVHD severity.

Role of GSPE in improving early cerebral vascular damage by inhibition of Profilin-1 expression in a ouabain-induced hypertension model.

OBJECTIVE: Grape seed proanthocyanidin extract (GSPE), as one of the most popular natural drug extracted from the grape, has been reported to improve endothelial function and arteriosclerosis. However, little is known about the influence of GSPE on hypertension and vascular remodeling. Profilin-1, an Actin-binding protein, is closely involved in the remodeling of large vessels in ouabain-induced hypertension. To date, there is no effective prevention or treatment in place for the high incidence of ischemic stroke associated with hypertension. In this study, we aimed to determine the role of GSPE via inhibition Profilin-1 in ischemic cerebral cortices of ouabain-hypertension rats and potentially provide a new target to prevent stroke associated with hypertension. MATERIALS AND METHODS: The blood pressure of male Sprague-Dawley (SD) rats was measured during a period of ouabain-induced hypertension. The expression of Profilin-1, vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS) in the cerebral cortex were determined by quantitative Real Time-PCR (qRT-PCR) and Western blot. Histopathological and behavioral tests were also conducted. RESULTS: Blood pressure elevation started at week 5 and reached clinical standards for hypertension at week 8. GSPE was proved to suppress Profilin-1 and VEGF levels through inhibition of Profilin-1-protein kinase B (AKT)-hypoxia inducible factor-1α (HIF-1α) signal pathway and promote eNOS expression. Moreover, the histopathological and ethiological improvement was observed in GSPE over-expression and Profilin-1 inhibition groups. CONCLUSIONS: We detected that GSPE could improve cerebral vascular damage through inhibiting Profilin-1 in an ouabain-induced hypertension model.

Overexpression of ezrin and galectin-3 as predictors of poor prognosis of cervical cancer.

The aim of this study was to explore the correlation of ezrin and galectin-3 expressions with prognosis in cervical cancer. The immunohistochemical method was applied to detect ezrin and galectin-3 expressions in normal cervix tissues (n=30), cervicitis tissues (n=28), cervical intraepithelial neoplasia (CIN) tissues (classified as I-III, n=89), and cervical carcinoma tissues (n=84). Follow-up was conducted for 5 to 78 months to analyze the correlation of protein expressions with prognosis. Ezrin and galectin-3 expressions in cervical cancer were significantly higher than in normal cervix, cervicitis and CIN (all P<0.05), and expressions in CIN were significantly higher than in normal cervix and cervicitis (both P<0.05). The expressions of ezrin and galectin-3 were both related with histological grade, deep myometrial invasion and lymph node metastasis (all P<0.05). Spearman analysis showed that ezrin expression was positively correlated with galectin-3 expression in cervical cancer (r=0.355, P<0.05). The survival rate of patients with high expressions of ezrin and galectin-3 was significantly lower than those with low expressions of proteins (both P<0.05). The expressions of ezrin and galectin-3, histological grade, depth of stromal invasion, and lymph node metastasis are risk factors affecting the survival rate of patients with cervical cancer. The expressions of ezrin and galectin-3 were correlated with the development of cervical cancer, and overexpressions of those proteins were indicative of poor prognosis in patients with cervical cancer.

Neutral Ceramidase Secreted Via Exosome Protects Against Palmitate-Induced Apoptosis in INS-1 Cells.

Aims: To investigate the effects of neutral ceramidase (NCDase) packaged in exosomes that are secreted from ß-cells on free fatty acid (FFA)-induced ß-cells apoptosis and its role in regulation of sphingolipid-mediated signaling pathway. Methods: HPLC and Western blotting were performed to determine NCDase activity and expression. Annexin V-fluorescein-isothiocyanate/propidium iodide flow cytometry was used to assess apoptosis. Electrospray ionization tandem mass spectrometry was used for ceramide (Cer), sphingosine-1-phosphate (S1P), and sphingosine (SPH) determination. Results: INS-1 cells over-expressed NCDase secreted active NCDase via exosomes. Exosomes isolated from the cultured medium of INS-1 cells that oxpressed NCDase could ameliorate palmitate-induced apoptosis. Furthermore, the results showed that exosome-derived NCDase treatment reduced intracellular Cer/S1P ratio. Conclusions: ß-cell secreted active NCDase via exosome, the exosome-packaged-NCDase protected ß-cells from FFA-induced apoptosis through regulating sphingolipid metabolites and it might be a potential treatment for ß-cell lipotoxicity and type 2 diabetes.

Roles of PECAM-1 in cell function and disease progression.

Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) belongs to immunoglobulin superfamily, which is key factor for adhesion and accumulation of platelets. It is proved that PECAM-1 is closely correlative with cell migration, proliferation, apoptosis, signal transduction and cellular immunity. Meanwhile, PECAM-1 involves in multiple clinical diseases, such as atherosclerosis, thrombosis and leukemia. This paper reviewed the structure and function of PECAM-1, and its roles in cell function and disease generation and progression.

Cancer-associated fibroblasts induce epithelial-mesenchymal transition of breast cancer cells through paracrine TGF-ß signalling.

BACKGROUND: Cancer-associated fibroblasts (CAFs) activated by tumour cells are the predominant type of stromal cells in breast cancer tissue. The reciprocal effect of CAFs on breast cancer cells and the underlying molecular mechanisms are not fully characterised. METHODS: Stromal fibroblasts were isolated from invasive breast cancer tissues and the conditioned medium of cultured CAFs (CAF-CM) was collected to culture the breast cancer cell lines MCF-7, T47D and MDA-MB-231. Neutralising antibody and small-molecule inhibitor were used to block the transforming growth factor-ß (TGF-ß) signalling derived from CAF-CM, which effect on breast cancer cells. RESULTS: The stromal fibroblasts isolated from breast cancer tissues showed CAF characteristics with high expression levels of α-smooth muscle actin and SDF1/CXCL12. The CAF-CM transformed breast cancer cell lines into more aggressive phenotypes, including enhanced cell-extracellular matrix adhesion, migration and invasion, and promoted epithelial-mesenchymal transition (EMT). Cancer-associated fibroblasts secreted more TGF-ß1 than TGF-ß2 and TGF-ß3, and activated the TGF-ß/Smad signalling pathway in breast cancer cells. The EMT phenotype of breast cancer cells induced by CAF-CM was reversed by blocking TGF-ß1 signalling. CONCLUSION: Cancer-associated fibroblasts promoted aggressive phenotypes of breast cancer cells through EMT induced by paracrine TGF-ß1. This might be a common mechanism for acquiring metastatic potential in breast cancer cells with different biological characteristics.

Cochlear implantation in a patient with severe cochlear hypoplasia.

OBJECTIVE: We report the case of a successful cochlear implantation in a patient with severe cochlear hypoplasia. CASE REPORT: The outcome of cochlear implantation is generally less favourable for patients with cochlear hypoplasia than for those with a normal cochlear structure. In the reported patient, part of the electrode array was inserted into the internal auditory canal. Nevertheless, the benefits following cochlear implantation seemed to outweigh the risks for this patient. CONCLUSION: Cochlear hypoplasia is not necessarily a contraindication for cochlear implantation.

Pitfalls in the management of monaural deafness.

OBJECTIVE: We report a patient who underwent cochlear implantation in an ear with long-term deafness, after an acoustic neuroma had been removed surgically from the other, hitherto good ear and the cochlear nerve had subsequently been resected to relieve severe tinnitus. METHOD: Case report. RESULTS: The patient could not tolerate the cochlear implant, because of a moderate headache due to the stimulation level necessary for environmental sound discrimination. CONCLUSION: Cochlear implantation in patients with long-term deafness should be considered carefully, even if deafness is monaural.

Changes in condylar and joint disc positions after bilateral sagittal split ramus osteotomy for correction of mandibular prognathism.

The effect of combined orthodontic and orthognathic treatment was studied retrospectively in 24 patients with skeletal class III malocclusions with mandibular hyperplasia, particularly the effect on temporomandibular joint (TMJ) disc position. The patients underwent preoperative orthodontic treatment, orthognathic surgery, and postoperative orthodontic treatment. The patients were studied clinically, radiographically with lateral cephalometric radiograph and MRI to locate the position of the TMJ disc in relation to the glenoid fossa. One patient had less pain after treatment, one lost abnormal joint clicking sounds after treatment. There were no TMJ symptoms in 20 of the 24 preoperatively and postoperatively. 48 sagittal MRI images showed that the disc length before treatment was 3.040-12.928 (mean 8.289+/-2.028) and after treatment was 3.699-11.589 (mean 8.097+/-1.966); results were not significant (p>0.05). Maximum disc displacement before treatment was 6.090 (mean 1.383), after treatment it was 11.931 (mean 2.193); results were not significant (p>0.05). The results suggest that combined orthodontic and orthognathic treatment (including bilateral SSRO and rigid internal fixation) can be used safely to correct skeletal class III malocclusion with mandibular hyperplasia without causing additional TMJ symptoms.

Effects of lipoxin A(4) on lipopolysaccharide induced proliferation and reactive oxygen species production in RAW264.7 macrophages through modulation of G-CSF secretion.

OBJECTIVE: To investigate the effects of lipoxin A(4) (LXA(4)) on lipopolysaccharide (LPS) induced proliferation and reactive oxygen species production in RAW264.7 macrophages. METHODS: RAW264.7 macrophages were treated with or without LPS in the absence or presence of LXA(4). In another experiment, the cells were incubated with rmG-CSF (recombinant mouse granulocyte-colony stimulating factor) or neutralizing anti-G-CSF Ab in the absence or presence of LPS and LXA(4). The proliferation effects were detected by Cell Counting Kit-8 assay. Reative oxygen species were quantified by flow cytometry and laser confocal scanning microscopy. G-CSF gene expression and protein secretion were measured by real time PCR and ELISA, respectively. IkappaBalpha degradation and NF-kappaB translocation were determined by Western blot, and NF-kappaB transcriptional activity was tested by transfections and luciferase activities assay. RESULTS: (1) LXA(4) controlled LPS-induced proliferation and reactive oxygen species production. (2) LXA(4) decreased LPS-induced G-CSF gene expression and protein secretion. (3) LXA(4) restrained LPS-induced IkappaBalpha degradation, NF-kappaB translocation, and NF-kappaB transcriptional activity. (4) rmG-CSF could rescue the inhibitory effects of LXA(4), and neutralizing anti-G-CSF Ab was able to enhance the roles of LXA(4). CONCLUSIONS: LXA(4) inhibited LPS-induced proliferation and reactive oxygen species production in RAW264.7 macrophages partially through modulation of G-CSF secretion.