Identification of FKBP10 prognostic value in lung adenocarcinoma patients with surgical resection of brain metastases: A retrospective single-institution cohort study

Abstract Objective To explore the expression levels and clinical value of FKBP10 in lung adenocarcinoma brain metastases. Design A retrospective single-institution cohort study. Patients The perioperative records of 71 patients with lung adenocarcinoma brain metastases who underwent surgical resection at the authors' institution between November 2012 and June 2019 were retrospectively analyzed. Methods The authors evaluated FKBP10 expression levels using immunohistochemistry in tissue arrays of these patients. Kaplan-Meier survival curves were constructed, and a Cox proportional hazards regression model was used to identify independent prognostic biomarkers. A public database was used to detect FKBP10 expression and its clinical value in primary lung adenocarcinoma. Results The authors found that the FKBP10 protein was selectively expressed in lung adenocarcinoma brain metastases. Survival analysis showed that FKBP10 expression (p = 0.02, HR = 2.472, 95% CI [1.156, 5.289]), target therapy (p < 0.01, HR = 0.186, 95% CI [0.073, 0.477]), and radiotherapy (p = 0.006, HR = 0.330, 95% CI [0.149, 0.731]) were independent prognostic factors for survival in lung adenocarcinoma patients with brain metastases. The authors also detected FKBP10 expression in primary lung adenocarcinoma using a public database, found that FKBP10 is also selectively expressed in primary lung adenocarcinoma, and affects the overall survival and disease-free survival of patients. Limitations The number of enrolled patients was relatively small and patients' treatment options varied. Conclusions A combination of surgical resection, adjuvant radiotherapy, and precise target therapy may benefit the survival of selected patients with lung adenocarcinoma brain metastases. FKBP10 is a novel biomarker for lung adenocarcinoma brain metastases, which is closely associated with survival time and may serve as a potential therapeutic target.

The role of macrophage polarization and interaction with renal tubular epithelial cells in ischemia-reperfusion induced acute kidney injury / 生理学报

Acute kidney injury (AKI) is a common critical clinical disease characterized by a sharp decline of renal function. Ischemia-reperfusion (IR) is one of the main causes of AKI. The mortality of AKI remains high due to the lack of early diagnosis and cause specific treatment. IR rapidly initiates innate immune responses, activates complement and innate immune cells, releasing a large number of injury-related molecules such as high mobility group box-1 (HMGB1), inflammatory mediators such as caspase-3, and then recruits immune inflammatory cells including M1 macrophages (Mϕ) to the microenvironment of injury, causing apoptosis and necrosis of renal tubular epithelial cells (TECs). Dead cells and associated inflammation further activate the adaptive immune system, which not only aggravates tissue damage, but also initiates M2 Mϕ participated inflammatory clearance, tissue repair and regeneration. Mϕ, professional phagocytes, and TECs, semi-professional phagocytes, can phagocytose around damaged cells including apoptotic Mϕ and TECs, which are key innate immune cells to regulate the outcome of injury, repair or fibrosis. In recent years, it has been found that erythropoietin (EPO) not only binds to the homodimeric receptor (EPOR)2 to induce erythropoiesis, but also binds to the heterodimeric receptor EPOR/βcR, also known as innate repair receptor, which plays renoprotective roles. Properdin is the only positive regulator in the complement activation of alternative pathway. It also can effectively identify and bind to early apoptotic T cells and facilitate phagocytic clearing by Mϕ through a non-complement activation-dependent mechanism. Our previous studies have shown that Mϕ and TECs associated with EPO and its receptors and properdin are involved in IR injury and repair, but the underlying mechanism needs to be further explored. As an important carrier of cell-to-cell signal transmission, exosomes participate in the occurrence and development of a variety of renal diseases. The role of exosomes involved in the interaction between Mϕ and TECs in IR-induced AKI is not fully defined. Based on the available results in the role of Mϕ and TECs in renal IR-induced AKI, this review discussed the role of Mϕ polarization and interaction with TECs in renal IR injury, as well as the participation of EPO and its receptors, properdin and exosomes.

Effect of mogrol on lipid metabolism of hepatocellular carcinoma HepG2 cells by activatingAMPK signaling pathway / 中国肿瘤生物治疗杂志

@# Objective： ：To study the regulatory effect of mogrol (MO) on lipid metabolism of hepatic cancer cells and its molecular mechanism. Methods: Oleic acid (OA) was used to induce fat accumulation in hepatocellular carcinoma HepG2 cells and to establish a steatosis cell model. CCK-8 method was used to detect the cytotoxicity of MO to HepG2 cells, and an experimental working concentration without obvious cytotoxicity of MO was chosen. After being treated with different concentrations of MO, lipid accumulation in the cells was observed by oil red O staining, and the contents of triglyceride (TG) and cholesterol (TC) in the cells were measured. Key genes involving in lipid metabolism were screened out by high-throughput transcriptome sequencing qPCR was used to detect the mRNA expressions of ,SREBP-1c and FASN, while Western Blot was used to detect the protein expressions of p-AMPKα, SREBP-1c and FASN in cells of model group and treatment group. Results: After OA induction, a large amount of lipids accumulated in HepG2 cells, the contents of TG and TC increased significantly. Three key genes (SREBP-1c, FASN and p-AMPK α) involving in lipid metabolism of hepatic cancer cells were screened out. After OA induction, the mRNA expressions of SREBP-1c and FASN increased, the protein expression of p-AMPK α decreased while the protein expressions of SREBP-1c, FASN and other proteins increased significantly. After intervention with working concentration of MO, intracellular lipid accumulation, contents of TG and TC, mRNA expressions of SREBP-1c, FASN and protein expressions of SREBP-1c, FASN decreased significantly, while the expression of p-AMPKα increased. Conclusion: Mogrol can inhibit the synthesis of fatty acids by activating the expression level of AMPK signaling pathway related factors SREBP-1c and FASN, so as to play the role of regulating lipid metabolism.

Mental Health Status Assessment in Polycystic Ovarian Syndrome Infertility Patients: A Pilot Study / 华中科技大学学报（医学）（英德文版）

Polycystic ovary syndrome (PCOS) is a major endocrine disorder afflicting women of reproductive age.Women with PCOS are more likely to suffer from mental health disturbances than healthy women.The "infertility" suffered by PCOS patients would also lead to mental health disturbances.Symptom Checklist 90 (SCL-90) and questionnaire which includes patients' socio-economic and demographic data were used to assess the mental health status of PCOS (n=103) and non-PCOS (n=ll0) infertile patients.Logistic regression analysis and t-tests were used for comparative analysis.The data demonstrated that scores of depression,interpersonal sensitivity,obsessive-compulsive,and hostility symptoms in PCOS infertile patients were significantly higher than those in the non-PCOS infertile patients (P＜0.05).Logistic regression analysis revealed that acne had negative effect on mental health status (P＜0.05).Secondary infertile PCOS patients were more easily to suffer from somatization,interpersonal sensitivity,obsessive-compulsive,anxiety,hostility and paranoid ideation symptoms than the primary infertile PCOS patients (P＜0.05).The results suggested that the PCOS patients especially the secondary infertile PCOS patients had obvious mental health disturbances.The ache might play an importance role in the occurrence of mental health disturbances in PCOS patients.PCOS related symptoms may be risk factors of mental health status in PCOS patients with infertility.More attention should be paid to the PCOS infertile patients,and mental health therapy should be considered if necessary.

Expression of E2A in Mid-secretory Endometrium of Women Suffering from Recurrent Miscarriage / 华中科技大学学报（医学）（英德文版）

E2A is involved in promoting forkhead box P3 (FOXP3) and retinoid-related orphan receptor gamma t (RORγt) gene transcription,which are pivotal transcription factors of T regulatory cells and Thl7 cells,respectively.Little is known about the involvement of E2A in pregnancy process.This study aimed to investigate the expression of E2A,cytotoxic T-lymphocyte-associated protein 4 (CTLA-4),and Foxp3 in luteal phase endometrium of women suffering recurrent miscarriage (RM) (n=21) and control group (n=11) by immunohistochemistry,with the Vectra(R) automated quantitative pathology imaging system for analysis.The percentage of E2A+ cells and CTLA-4+ cells was significantly higher in the endometrium of women with RM than in the controls.There was positive correlation between E2A and CTLA-4 (r=0.523,P=0.002),E2A and FOXP3 (r=0.380,P=0.032),and FOXP3 and CTLA-4 (r=0.625,P=0.000) in the mid-secretory phase of endometrium for all subjects.It was concluded that the abnormal expression of endometrial E2A existed in mid-secretory endometrium of women with RM,and there was a positive correlation between E2A and FOXP3,and E2A and CTLA-4,suggesting the possible regulation role of E2A involved in regulating endometrium receptivity.

ATP Synthase β-subunit Abnormality in Pancreas Islets of Rats with Polycystic Ovary Syndrome and Type 2 Diabetes Mellitus / 华中科技大学学报（医学）（英德文版）

This study investigated the abnormal expression of ATP synthase β-subunit (ATPsyn-β) in pancreas islets of rat model of polycystic ovary syndrome (PCOS) with type 2 diabetes mellitus (T2DM),and the secretion function changes after up-regulation of ATP5b.Sixty female SD rats were divided into three groups randomly and equally.The rat model of PCOS with T2DM was established by free access to the high-carbohydrate/high-fat diet,subcutaneous injections of DHEA,and a single injection of streptozotocin.The pancreas was removed for the detection of the ATPsyn-β expression by immunohistochemical staining,Western blotting and reverse transcription-PCR (RT-PCR).The pancreas islets of the rats were cultured,isolated with collagenase Ⅴ and purified by gradient centrifugation,and the insulin secretion after treatment with different glucose concentrations was tested.Lentivirus ATP5b was successfully constructed with the vector of GV208 and transfected into the pancreas islets for the over-expression of ATPsyn-β.The insulin secretion and intracellular ATP content were determined after transfection of the PCOS-T2DM pancreas islets with Lenti-ATP5b.The results showed that the expression of ATPsyn-β protein and mRNA was significantly decreased in the pancreas of PCOS-T2DM rats.The ATP content in the pancreas islets was greatly increased and the insulin secretion was improved after the up-regulation of ATPsyn-β in the pancreas islets transfected with lenti-ATP5b.These results indicated that for PCOS,the ATPsyn-β might be one of the key factors for the attack of T2DM.

Effects and molecular mechanism of chitosan-coated levodopa nanoliposomes on behavior of dyskinesia rats

BACKGROUND: Chitosan, the N-deacetylated derivative of chitin, is a cationic polyelectrolyte due to the presence of amino groups, one of the few occurring in nature. The use of chitosan in protein and drug delivery systems is being actively researched and reported in the literature RESULTS: In this study, we used chitosan-coated levodopa liposomes to investigate the behavioral character and the expression of phosphorylated extracellular signal-regulated kinase (ERK1/2), dopamine- and cAMP-regulated phos-phoprotein of 32 kDa (DARPP-32) and FosB/AFosB in striatum of rat model of levodopa-induced dyskinesia (LID). We found that scores of abnormal involuntary movement (AIM) decreased significantly in liposome group (P < 0.05), compared with levodopa group. Levels of phospho-ERK1/2, phospho-Thr34 DARPP-32 and FosB/AFosB in striatum decreased significantly in liposome group lesion side compared with levodopa group (P < 0.05). However, both of two groups above have significantly differences compared with the control group (P < 0.05). CONCLUSION: Chitosan-coated levodopa liposomes may be useful in reducing dyskinesias inducing for Parkinson disease. The mechanism might be involved the pathway of signaling molecular phospho-ERK1/2, phospho-Thr34 DARPP-32 and AFosB in striatum

Down-regulated clusterin expression enhances sensitivity of hepatoma cells to anti-cancer drugs / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the relationship between and underlying mechanistic pathway of clusterin (CLU) and chemo-resistance ofhepatocellular carcinoma (HCC) cells.</p><p><b>METHODS</b>CLU protein expression in HCC cell lines (Hep3B, SMMC7721, PLC, and HepG2) and HepG2/ADM cells was quantified by western blotting. Four short-hairpin (sh)RNAs designed to block CLU-mRNA were generated, screened by RT-PCR, and transfected into the cells to determine effects of CLU on cell viability and apoptosis. Effects of CLU blockade on drug efflux pump activity were measured by flow cytometry.</p><p><b>RESULTS</b>CLU was found to be over-expressed in HCC cell lines and HepG2/ADM cells. The four shRNAs inhibited CLU-mRNA as follows (vs. levels in untransfected cells): shRNA-1: 73.68% (q =23.011, P < 0.01), shRNA-2: 39.26% (q =11.991, P < 0.01), shRNA-3: 62.36% (q =19.392, P < 0.01), and shRNA-4: 55.35% (q =17.149, P < 0.01). shRNA-mediated depletion of CLU led to increased sensitivity to anti-cancer drugs and increased doxorubicin-induced apoptosis in HepG2/ADM cells, as evidenced by the apoptosis ratio of the shRNA-1 group of 39.28% vs. the apoptosis ratio of the untransfected control group of 4.92%. Silencing of CLU also decreased drug etflux pump activity, and the level of MDR1/P-gp expression was significantly reduced (shRNA-1 group vs.untransfected control group: q =14.604, P < 0.01).</p><p><b>CONCLUSION</b>CLU repression may enhance sensitivity of HCC cells to anti-cancers drugs and represents a potential molecular-target for reversal of multidrug-resistant HCC.</p>

Effect of siRNA-mediated inhibition of nuclear transcription factor-kappa B on apoptosis of hepatocarcinoma cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of siRNA-mediated inhibition of NF-κB on apoptosis of hepatocarcinoma cells.</p><p><b>METHODS</b>Specific small interfering RNA Targeting NF-κB gene was synthesized and transfected into HepG2 cells by liposomes. Nested RT-PCR and quantitative RT-PCR were used to detect the mRNA expression of NF-κB. Immunohistochemistry, enzyme-linked immunosorbent assay and Western blot were performed to examine the protein expression of NF-κB. Annexin V-FITC was used to test cell apoptosis.</p><p><b>RESULTS</b>The expression of NF-κB in HepG2 cells (1.13+/-0.03) was significantly higher (t=27.02, P<0.05) than that in normal hepatocytes (0.29+/-0.07). The down-regulation of NF-κB expression was depended on the dosage of siRNA and the time after transfection. 72 h after siRNA transfection, NF-κB expression reduced by 93% and 62% at the mRNA and protein levels, respectively. The apoptosis of HepG2 cells increased by 85% with NF-κB inhibition.</p><p><b>CONCLUSIONS</b>NF-κB is abnormally active in HepG2 cells and NF-κB inhibition mediated by siRNA promotes HepG2 cells apoptosis. It suggested that NF-κB could be a potential target for HCC prevention gene therapy.</p>

Expression characteristics of hypoxia inducible factor-1a and its clinical values in hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the dynamic expression of hypoxia inducible factor-1alpha (HIF-1alpha) and its clinical values in hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The dynamic changes of liver pathology, HIF-1alpha transcription and expression were observed through the hepatoma model. The self-control specimens from 35 human HCC patients were collected and the expression, cellular distribution, and clinicopathological features of HIF-1alpha and its gene was analyzed by immunohistochemistry, western blotting and nested- PCR, respectively.</p><p><b>RESULTS</b>Both levels of hepatic HIF-1alpha and HIF-1alpha mRNA expression increased during the HCC development course. The incidence of HIF-1alpha and the ratio of HIF-1alpha to beta-actin was 0% and 0.16+/-0.02 in the control rats, 77.8% and 0.29+/-0.04 in the denatured rats, 88.9% and 0.52+/-0.03 in the precancerous rats, and 100% and 0.84+/-0.02 in the cancerous rats respectively, with significant difference between the control group and any of the experimental groups (P = 0.000). The positive HIF-1alpha was brown and granule-like and mainly presented in cytoplasm and few in nucleus. The incidence of HIF-1alpha was 80% (28/35) in HCC and 100% (35/35) in its surrounding tissues. The clinical pathological features indicated HIF-1alpha expression associated with tumor size and differentiation degree the of HCC. No correlation was found between HIF-1alpha and tumor numbers or positive-HBsAg.</p><p><b>CONCLUSIONS</b>HIF-1alpha expression is associated with occurrence and development of HCC, and is perhaps a target molecule for HCC therapy.</p>

Sequence analysis on drug-resistant gene of rpoB in Mycobacterium tuberculosis L-forms among pneumocoulosis patients complicated with tuberculosis / 中华流行病学杂志

Objective To study the drug-resistant characteristics genetic mutation of rpoB in Mycobacterium tuberculosis L-forms among patients of pneumoconiosis complicated with pulmonary tuberculosis. Methods A total of 42 clinical isolated strains of Mycobacterium tuberculosis L-forms were collected, including 31 drug-resistant strains. Their genomes DNA were extracted and target genes amplified by PCR. Hot regions in the rpoB gene were analyzed by automated DNA sequenator. Results No mutation of rpoB was identified in 11 rifampicin-sensitive strains while conformation changes were fotmd in 31 rifampicin-resistant strains. The mutation rate was 93.55% (29/31) in resistant strains, mainly concentrated in codon 531 (51.6%, 16/31) and 526 (32.26%, 10/31), happened base substitutions, including 27 unit point mutation and 2 two point mutation. The newly found mutation of codon 516 had not been reported by internal or overseas scholars. Conclusion The substitution of highly conserved amino acids encoded by rpoB gene resulted in the molecular mechanism was responsible for RFP resistance in Mycobacterium tuberculosis L-forms. It also proved that rpoB gene was in diversiform.