Identification of differentially expressed mRNAs as novel predictive biomarkers for gastric cancer diagnosis and prognosis.

BACKGROUND: Gastric cancer (GC) has a high mortality rate worldwide. Despite significant progress in GC diagnosis and treatment, the prognosis for affected patients still remains unfavorable. AIM: To identify important candidate genes related to the development of GC and identify potential pathogenic mechanisms through comprehensive bioinformatics analysis. METHODS: The Gene Expression Omnibus database was used to obtain the GSE183136 dataset, which includes a total of 135 GC samples. The limma package in R software was employed to identify differentially expressed genes (DEGs). Thereafter, enrichment analyses of Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were performed for the gene modules using the clusterProfile package in R software. The protein-protein interaction (PPI) networks of target genes were constructed using STRING and visualized by Cytoscape software. The common hub genes that emerged in the cohort of DEGs that was retrieved from the GEPIA database were then screened using a Venn Diagram. The expression levels of these overlapping genes in stomach adenocarcinoma samples and non-tumor samples and their association with prognosis in GC patients were also obtained from the GEPIA database and Kaplan-Meier curves. Moreover, real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting were performed to determine the mRNA and protein levels of glutamic-pyruvic transaminase (GPT) in GC and normal immortalized cell lines. In addition, cell viability, cell cycle distribution, migration and invasion were evaluated by cell counting kit-8, flow cytometry and transwell assays. Furthermore, we also conducted a retrospective analysis on 70 GC patients diagnosed and surgically treated in Wenzhou Central Hospital, Dingli Clinical College of Wenzhou Medical University, The Second Affiliated Hospital of Shanghai University between January 2017 to December 2020. The tumor and adjacent normal samples were collected from the patients to determine the potential association between the expression level of GPT and the clinical as well as pathological features of GC patients. RESULTS: We selected 19214 genes from the GSE183136 dataset, among which there were 250 downregulated genes and 401 upregulated genes in the tumor samples of stage III-IV in comparison to those in tumor samples of stage I-II with a P-value < 0.05. In addition, GO and KEGG results revealed that the various upregulated DEGs were mainly enriched in plasma membrane and neuroactive ligand-receptor interaction, whereas the downregulated DEGs were primarily enriched in cytosol and pancreatic secretion, vascular smooth muscle contraction and biosynthesis of the different cofactors. Furthermore, PPI networks were constructed based on the various upregulated and downregulated genes, and there were a total 15 upregulated and 10 downregulated hub genes. After a comprehensive analysis, several hub genes, including runt-related transcription factor 2 (RUNX2), salmonella pathogenicity island 1 (SPI1), lysyl oxidase (LOX), fibrillin 1 (FBN1) and GPT, displayed prognostic values. Interestingly, it was observed that GPT was downregulated in GC cells and its upregulation could suppress the malignant phenotypes of GC cells. Furthermore, the expression level of GPT was found to be associated with age, lymph node metastasis, pathological staging and distant metastasis (P < 0.05). CONCLUSION: RUNX2, SPI1, LOX, FBN1 and GPT were identified key hub genes in GC by bioinformatics analysis. GPT was significantly associated with the prognosis of GC, and its upregulation can effectively inhibit the proliferative, migrative and invasive capabilities of GC cells.

[Observation of early clinical efficacy of unilateral biportal endoscopy technique in the treatment of lumbar postoperative adjacent segmental diseases].

OBJECTIVE: To investigate early clinical efficacy of unilateral biportal endoscopy technique for the treatment of lumbar postoperative adjacent segmental diseases. METHODS: Fourteen patients with lumbar postoperative adjacent segmental diseases were treated with unilateral biportal endoscopy technique from June 2019 to June 2020. Among them, there were 9 males and 5 females, aged from 52 to 73 years old, and the interval between primary and revision operations ranged from 19 to 64 months. Adjacent segmental degeneration occurred after lumbar fusion in 10 patients and after lumbar nonfusion fixation in 4 patients. All the patients received unilateral biportal endoscopy assisted posterior unilateral lamina decompression or unilateral approach to the contralateral decompression. The operation time, postoperative hospital stay and complications were observed. The visual analogue scale (VAS) of low back pain and leg pain, Oswestry Disability Index (ODI), modified Japanese Orthopaedic Association (mJOA) score were recorded before operation and at 3 days, 3 months, and 6 months after operation. RESULTS: All procedures were successfully completed. Surgical duration ranged from 32 to 151 min. Postoperative CT showed adequate decompression and preservation of most joints. Out of bed walking 1 to 3 days after surgery, postoperative hospital stay was 1 to 8 days, and postoperative follow-up was 6 to 11 months. All 14 patients returned to normal life within 3 weeks after surgery, and VAS, ODI, and mJOA scores improved significantly at 3 days and 3, 6 months after surgery. One patient occurred cerebrospinal fluid leak after operation, received local compression suture, and the wound healed after conservative treatment. One patient occurred postoperative cauda equina neurologic deficit, which was gradually recovered about 1 month after rehabilitation therapy. One patients advented transient pain of lower limbs after surgery, and the symptoms were relieved after 7 days of treatment with hormones, dehydration drugs and symptomatic management. CONCLUSION: Unilateral biportal endoscopy technique has a good early clinical efficacy in the treatment of lumbar postoperative adjacent segmental diseases, which may provide a new minimally invasive, non-fixation option for the treatment of adjacent segment disease.

JP1, a polypeptide specifically targeting integrin αVß3, ameliorates choroidal neovascularization and diabetic retinopathy in mice.

Anti-vascular endothelial growth factor (VEGF) drugs have revolutionized the treatment of neovascular eye diseases, but responses are incomplete in some patients. Recent evidence shows that integrins are involved in the pathogenesis of neovascular age-related macular degeneration and diabetic retinopathy. JP1, derived from an optimized seven-amino-acid fragment of JWA protein, is a polypeptide specifically targeting integrin αVß3. In this study we evaluated the efficacy of JP1 on laser-induced choroidal neovascularization (CNV) and retinal vascular leakage. CNV mice received a single intravitreal (IVT) injection of JP1 (10, 20, 40 µg) or ranibizumab (RBZ, 10 µg). We showed that JP1 injection dose-dependently inhibited laser-induced CNV; the effect of RBZ was comparable to that of 20 µg JP1; a combined IVT injection of JP1 (20 µg) and RBZ (5 µg) exerted a synergistic effect on CNV. In the 3rd month after streptozotocin injection, diabetic mice receiving IVT injection of JP1 (40 µg) or RBZ (10 µg) once a week for 4 weeks showed significantly suppressed retinal vascular leakage. In both in vivo and in vitro experiments, JP1 counteracted oxidative stress and inflammation via inhibiting ROS/NF-κB signaling in microglial cells, and angiogenesis via modulating MEK1/2-SP1-integrin αVß3 and TRIM25-SP1-MMP2 axes in vascular endothelial cells. In addition, intraperitoneal injection of JP1 (1, 5 or 10 mg) once every other day for 3 times also dose-dependently inhibited CNV. After intraperitoneal injection of FITC-labeled JP1 (FITC-JP1) or FITC in laser-induced CNV mice, the fluorescence intensity in the CNV lesion was markedly increased in FITC-JP1 group, compared with that in FITC group, confirming that JP1 could penetrate the blood-retinal barrier to target CNV lesion. We conclude that JP1 can be used to design novel CNV-targeting therapeutic agents that may replace current invasive intraocular injections.

Unexpected difficult airway due to severe upper tracheal distortion: A case report.

BACKGROUND: Difficult airway is a significant cause of anesthesia-associated death and disability. Currently, physical examinations of thyromental distance, mouth opening, Mafampaii classification, etc. combined with X-ray, computed tomography (CT), and other imaging technologies are mainly used to evaluate difficult airways. However, in many special cases, i.e., emergency surgery, imaging examinations cannot be completed preoperatively. Such patients' airway can only be evaluated through general physical examination, which inevitably increases the likelihood of an unexpected difficult airway during anesthesia. CASE SUMMARY: We report a rare case of difficult intubation because of severe upper trachea distortion after induction. Emergency holmium laser lithotripsy was performed under transurethral ureteroscopy because the patient had anuria for 4 d and a creatinine level of 890 µmol/L. Due to the urgency of the condition, chest radiography or chest CT was not examined before surgery and the anesthesiologist did not evaluate the airway adequately, resulting in an unexpected difficult airway. CONCLUSION: The incidence of tracheal malformation and tracheal stenosis is extremely low, but the risk of hypoxia and even death due to difficult airways is extremely high for such patients. It is recommended to complete preoperative imaging examinations of the airway. For life-threatening emergency patients, a pre-anesthesia reassessment should be performed and surgeons should be prepared to prevent and manage the difficult airway.

[Finite element analysis of the effect of plate placement angle on cervical spine stability in anterior cervical fusion surgery].

OBJECTIVE: Using the method of finite element analysis, to compare the biomechanical properties between the plate deviating from the long axis of the cervical spine and the standard placement of the plate in the anterior cervical fusion surgery. METHODS: A healthy female volunteer was selected and CT scan (C1-T1) was performed. Using Mimics 19.0, Geomagic Studio 2015, Solidworks 2018, Ansys Workbench 17.2 to establish a lower cervical spine (C3-C7) model and to verify the reliability of the model. Subsequently, anterior cervical plates of different angles and lengths were placed to establish an anterior cervical discectomy fusion (ACDF) model. Applying 73.6 N axial pressure and 1 NM pure moment on C3 to make the model produce flexion, extension, lateral bending and rotation activities, observed the model stress cloud diagram and recorded the maximum stress value of the instrument and the intervertebral mobility. RESULTS: The lower cervical spine (C3-C7) finite element model was established and verified against the published literature on the range of motion (ROM) of cervical spine. Effect of steel plate offset axis on stress distribution, maximum stress value and intervertebral ROM of internal fixation apparatus was minimal, and the mechanical effect of steel plate offset was less in double section steel plate than in single section steel plate. CONCLUSION: Little effect on the mechanical stability of the cervical spine was anticipated when the anterior cervical plate was not perfectly aligned with the long axis of the cervical spine. If the tilt of the plate in clinical surgery is less than 20°, there is no need to readjust the position of the plate.

JP3, an antiangiogenic peptide, inhibits growth and metastasis of gastric cancer through TRIM25/SP1/MMP2 axis.

BACKGROUND: Gastric cancer (GC) is the most prevalent gastrointestinal tumor with an unfavorable clinical prognosis. GC patients are largely threatened owing to metastasis and drug resistance. Tumor angiogenesis plays an important role in the development of gastric cancer and is a challenge in the treatment of gastric cancer. METHODS: Mouse xenograft models were used for screening of therapeutic peptides on GC growth and metastasis. Routine laboratory experimental methods including conditional cell culture, tube formation assay, qRT-PCR, Western blotting, immunohistochemistry (IHC), ubiquitination assay, and immunofluorescence (IF) were used in mechanism investigation; protein docking analysis and coimmunoprecipitation (Co-IP) were used for prediction and confirmation of interactions between JP3/SP1 and TRIM25/MEK1/2. RESULTS: We identified an MMP2-targeted peptide JP3 that plays inhibiting roles in modulating growth and metastasis of GC in vivo and has no observable toxic side effects. JP3 reduced tumor microvessel density (MVD) in vivo and human umbilical vein endothelial cells (HUVECs) tube formation in vitro. Mechanistic studies revealed that JP3 reduces polyubiquitination-mediated degradation of TRIM25 by increasing the stability of TRIM25 through phosphorylating it at Ser12. TRIM25, as an E3 ubiquitin ligase, promoted the ubiquitin of SP1 at K610, further suppressed expression of MMP2 and inhibited angiogenesis in GC. Importantly, the inversely association between TRIM25 and SP1 protein level was further verified in human GC tissues. Decreased TRIM25 expression and increased SP1 expression in tumor tissues were positively correlated with poor prognosis of GC patients. CONCLUSIONS: MMP2-targeted peptide JP3 plays a therapeutic role in GC through anti-angiogenesis by modulating TRIM25/SP1/MMP2.

Clinical features and prognostic factors in patients with bone metastases from non-small cell lung cancer.

OBJECTIVE: To investigate the clinical features and evaluate the prognostic factors in patients with bone metastases from non-small cell lung cancer (NSCLC). METHODS: We retrospectively investigated 356 patients with NSCLC with bone metastases from January 2012 to December 2017. The overall survival (OS) and 1-year survival rate were calculated by Kaplan-Meier analysis and compared by univariate analysis using the log-rank test. Multivariate analysis was performed using the Cox proportional hazards model. RESULTS: A total of 694 sites of bone metastases were determined among the 356 patients. The most common site of bone metastases was the ribs. The median OS was 12.5 months and the 1-year survival was 50.8% in the overall population. Univariate analysis revealed that histological type, number of bone metastases, Eastern Cooperative Oncology Group performance status (ECOG PS), bisphosphonate therapy, and serum calcium, lactate dehydrogenase, and alkaline phosphatase were significantly correlated with prognosis. Multivariate analysis identified multiple bone metastases, ECOG PS ≥2, lactate dehydrogenase ≥225 U/L, and alkaline phosphatase ≥140 U/L as independent negative prognostic factors. CONCLUSION: Multiple bone metastases, high ECOG PS, and high serum alkaline phosphatase and lactate dehydrogenase are independent negative prognostic factors for bone metastases from NSCLC.

Transforaminal Endoscopic Discectomy and Foraminoplasty for Treating Central Lumbar Stenosis.

OBJECTIVE: Lumbar spinal stenosis is a medical condition characterized by the narrowing of the spinal canal as a consequence of bone and soft tissue degeneration, including disc herniation, facet and ligamentum flavum hypertrophy, and osteophyte formation. The percutaneous transforaminal endoscopic discectomy (PTED) technique is one of the emerging surgical alternatives for treating central lumbar stenosis. The present study aims to describe the present techniques of PTED and foraminoplasty for central lumbar stenosis, and discuss the feasibility and advantages of this technique. METHODS: A total of 55 patients with an average age of 50 years were recruited in this study. They were operated on between August 2017 and June 2018 by a single surgeon for symptomatic lumbar stenosis using the PTED and foraminoplasty technique, along with a detailed description of the present technique. The retrospective analysis of 55 patients operated between August 2017 and June 2018 by a single surgeon for symptomatic lumbar stenosis using the PTED and foraminoplasty techniques, and the detailed description of the present technique were the focus of the present study. For all patients, the PTED and foraminoplasty procedure was performed under local anesthesia in the lateral position on a radiolucent table using C-arm fluoroscopy. The retrospective analysis evaluated the outcomes of symptoms through follow-up interviews at six weeks, six months, and one year after surgery. The analyzed parameters included surgery time, intraoperative blood loss, postoperative complications, visual analog scale (VAS) score, Japan Orthopedic Association (JOA) score, and the Oswestry Disability Index (ODI). The modified MacNab criteria were adopted. RESULTS: The average duration of symptoms was 15.6 weeks. The mean operative time was 161 minutes. The mean volume of intraoperative blood loss was 21 mL. The mean follow-up period was 14.6 months. The average preoperative VAS score for leg pain and low back pain was 6.8 and 5.5, respectively. The preoperative ODI and JOA score was 49.2 and 14.6, respectively. At the final follow-up, all 55 patients had an average VAS score of 1.1 for leg pain and 0.5 for low back pain. At the same time, the average ODI and JOA score was seven and 24.5, respectively. The statistical analysis showed that the VAS score, ODI value, and JOA score were significantly lower in all time-points at post-operation, when compared to those at pre-operation. For the modified MacNab criteria, the final outcome results were excellent in 39 patients (70.9%), good in nine patients (16.4%), fair in four patients (7.3%), and poor in two patients (3.6%), and the overall success rate was 89.1%. Two patients underwent a second operation during the follow-up period, and their symptoms were released after the reoperation. CONCLUSION: PTED and foraminoplasty technique showed promising outcomes in the treatment of central lumbar stenosis in a 1-year follow-up period. It suggested that PTED and foraminoplasty might be applied as a safe and effective therapeutic option for patients with lumbar stenosis.

Clinical assessment of magnetic resonance imaging-guided radiofrequency ablation for breast cancer.

The aim of the present study was to investigate the efficacy of magnetic resonance imaging (MRI)-guided radiofrequency ablation (RFA) for breast cancer patients who cannot undergo traditional surgery. A total of 10 patients were treated by MRI-guided RFA, of whom 6 had stage IV disease (lung metastasis n=3, bone metastasis n=1, liver metastasis n=1 and mediastinal metastasis n=1) and the remaining 4 patients, who refused surgery, had stage III disease accompanied by severe underlying conditions. The changes in feasibility, tumor volume, bleeding, local recurrence, metastasis and complications were evaluated after RFA. The patients were followed up at 1, 3, 6 and 12 months, and annually thereafter. A total of 14 RFA sessions were successfully performed (100%) in the 10 patients, among whom 7 patients underwent a single RFA session, 2 patients underwent two sessions, and 1 patient underwent three sessions. Compared with pre-RFA, the volume of the tumors at 6 months after RFA was markedly decreased. There was no local tumor recurrence or metastasis detected during a mean follow-up period of 19.5±3.46 months, and no major complications were reported. Therefore, RFA was found to be a minimally invasive and feasible treatment method in the present study, and MRI-guided RFA may be a promising alternative option for breast cancer patients who are unable to tolerate surgery. However, more prospective studies on the applicability of RFA in breast cancer are required.

T-cell lymphomas associated gene expression signature: Bioinformatics analysis based on gene expression Omnibus.

OBJECTIVES: Due to the low incidence and the heterogeneity of subtypes, the biological process of T-cell lymphomas is largely unknown. Although many genes have been detected in T-cell lymphomas, the role of these genes in biological process of T-cell lymphomas was not further analyzed. METHODS: Two qualified datasets were downloaded from Gene Expression Omnibus database. The biological functions of differentially expressed genes were evaluated by gene ontology enrichment and KEGG pathway analysis. The network for intersection genes was constructed by the cytoscape v3.0 software. Kaplan-Meier survival curves and log-rank test were employed to assess the association between differentially expressed genes and clinical characters. RESULTS: The intersection mRNAs were proved to be associated with fundamental processes of T-cell lymphoma cells. These intersection mRNAs were involved in the activation of some cancer-related pathways, including PI3K/AKT, Ras, JAK-STAT, and NF-kappa B signaling pathway. PDGFRA, CXCL12, and CCL19 were the most significant central genes in the signal-net analysis. The results of survival analysis are not entirely credible. CONCLUSIONS: Our findings uncovered aberrantly expressed genes and a complex RNA signal network in T-cell lymphomas and indicated cancer-related pathways involved in disease initiation and progression, providing a new insight for biotargeted therapy in T-cell lymphomas.

Dietary supplements during the cold season increase rumen microbial abundance and improve rumen epithelium development in Tibetan sheep.

Livestock on the Qinghai-Tibetan Plateau are faced with extreme harsh winters and are often in negative energy balance during this period. Dietary supplementation can improve growth performance of Tibetan sheep and, consequently, we hypothesized that it would also increase microbial abundance and rumen epithelium development. To test this hypothesis, we examined the effect of feed supplementation during the cold season on rumen microbes, fermentation, epithelium development, and absorptive capability in Tibetan sheep. Eighteen 1-yr-old ewes (BW = 29.4 ± 1.79, kg) were offered oat hay ad libitum for 60 d and divided randomly into three groups: 1) no supplement; control group (CON); 2) urea-molasses lick block supplement (BS); and 3) concentrate feed supplement (CS). The ADG of CS ewes (143.3, g/d) was greater (P < 0.05) than BS ewes (87.9, g/d), which was greater (P < 0.05) than CON ewes (44.5, g/d). Serum concentrations of GH, IGF-1, and IGF-2 in the CS and BS groups were greater than in the CON group (P < 0.05). Greater relative abundance of protozoa, Ruminococcus albus, Fibrobacter succinogenes, Streptococcus bovis, and Ruminobacter amylophilus was observed in the CS and BS groups than in the CON group (P < 0.05), and relative abundances of rumen fungi, Butyrivibrio fibrisolvens, and Prevotella ruminicola in the CS group were greater than in the BS and CON groups (P < 0.05). Ruminal total VFA, ammonia, and microbial protein concentrations in the CS and BS groups were greater than in the CON group (P < 0.05), and in the CS group were greater than in the BS group (P < 0.05). Ruminal papillae width and surface area in the CS and BS groups were greater than in the CON group (P < 0.05), while in the CS group were greater than in the BS group (P < 0.05). The mRNA expressions of IGFBP5, NHE1 (sodium/hydrogen antiporter, isoform 1), DRA (downregulated in adenoma), and Na+/K+-ATPase (sodium/potassium ATPase pump) in ruminal epithelium were greater in the CS and BS groups than in the CON group (P < 0.05), and in the CS group was greater than in the BS group (P < 0.05), while NHE3 (sodium/hydrogen antiporter, isoform 3), MCT1 (monocarboxylate transporter 1), and MCT4 (monocarboxylate transporter 4) mRNA expressions in the CS group were greater than in the BS and CON groups (P < 0.05). It was concluded that supplementing Tibetan sheep during the cold season increases rumen microbial abundance and improves fermentation parameters, rumen epithelium development, and absorptive capability.

FBI-1 mRNA in normal mucosa is an independent prognostic factor in colorectal cancer patients.

Although several studies provide evidence that FBI-1 is an important gene regulator in colorectal cancer (CRC), it is noteworthy that, to our knowledge, no analysis of the correlation between FBI-1 expression and prognosis in CRC has been reported. Using real-time RT-PCR, we detected FBI-1 mRNA in 161 CRC patients (primary tumor, along with the corresponding normal mucosa), 36 liver metastases, and analyzed the relationship of its expression with clinicopathological features. Colon cancer cell lines were used to study FBI-1 function. Our study found that FBI-1 was significant up-regulated in tumor tissue (2.621 ± 0.157) compared with the corresponding normal mucosa (1.620 ± 0.165, P < 0.0001). FBI-1 in normal mucosa was a prognostic factor (P = 0.039, RR 0.431, 95% CI 0.194-0.958), independent of gender, age, stage, and differentiation. High levels of FBI-1 mRNA were related with good survival. Patients with complications had a higher primary tumor FBI-1 expression than those without complications (3.400 ± 0.332 vs. 2.516 ± 0.241, P = 0.032). Suppression of FBI-1 in colon cancer cell lines could repress proliferation of cancer cells. In conclusion, FBI-1 mRNA is overexpressed in CRC, and takes part in the development of CRC. FBI-1 mRNA in normal mucosa is an independent prognostic factor. Our findings give further support to the concept of "field cancerization", and hint that when we study a biomarker, we should not only focus on the tumor tissue but also the corresponding normal mucosa.

[Significance of Smad4, Smad7 and Caveolin-1 expression in oral mucosa carcinogenesis of Wistar rats].

PURPOSE: To investigate the expression of Smad4, Smad7 and Caveolin-1 in the process of carcinogenesis of oral mucosa in Wistar rats, and to understand the changes of TGF-ß/Smad signaling pathway and Caveolin-1 in oral cancer. METHODS: Palatal mucosal carcinogenesis specimens of Wistar rats were obtained from School of Stomatology, Zhengzhou University, which included 5 samples of normal mucosa, 10 samples of simple hyperplasia mucosa, 6 samples of mild mucosal dysplasia, 7 samples of moderate mucosal dysplasia, 13 samples of mucosa severe mucosal dysplasia, and 28 samples of oral cancer tissue. The expression of Smad4, Smad7 and Caveolin-1 was detected by immunohistochemistry. SPSS15.0 software package was used for statistical analysis. RESULTS: The expression of Smad4 decreased in normal and hyperplastic epithelia, dysplasticepithelia and oral cancer gradually, the difference of the expression among the three groups was significant (P<0.05). The expression of Smad7 and Caveolin-1 increased in normal and hyperplastic epithelia, dysplasticepithelia and oral cancer gradually, respectively; the difference of the expression among the 3 groups was significant (P<0.05). Spearman correlation analysis showed that Smad4 was negatively correlated with Smad7, Smad4 was negatively correlated with caveolin-1, Smad7 was positively correlated with Caveolin-1 (P<0.05). CONCLUSIONS: Synergistic effects may exist among Smad4, Smad7 and caveolin-1 in carcinogenesis of oral cancer.

GSK-3ß suppresses HCC cell dissociation in vitro by upregulating epithelial junction proteins and inhibiting Wnt/ß-catenin signaling pathway.

Glycogen synthase kinase-3ß (GSK-3ß) is required in the expression of epithelial junction proteins. It was found downregulated in hepatocellular carcinoma (HCC) tissues. The purpose of this study was to investigate the role of GSK-3ß in modulating the metastatic behaviors of human HCC cell lines in vitro. In this study, the expression level of GSK-3ß was measured in 4 human HCC cell lines, and the small interfering RNA (siRNA) vectors against or plasmids encoding GSK-3ß were used to evaluate the responses of target cells to the knockdown or overexpression of this kinase, respectively. Our results showed that GSK-3ß expression was significantly lower in human HCC cell lines with high metastatic potential than that in HCC cell lines without metastatic characteristics or in a normal human liver cell line. The knockdown of GSK-3ß by siRNA led to a decreased expression of the epithelial junction molecules (ZO-1, E-cadherin) and an increase in the expression of a mesenchymal cell marker (α-SMA) and a gene transcription factor (ß-catenin), resulting in enhanced tumor cell dissemination. In contrast, gain-of-function studies revealed that ectopic expression of GSK-3ß reduced invasive and migratory abilities of HCC cells accompanied by decreased HCC cell proliferation and induced apoptosis. More importantly, downregulation of GSK-3ß led to an increase in the expression and accumulation of ß-catenin in the nuclei, promoting gene transcription. In conclusion, GSK-3ß might play a vital role in suppressing HCC dissociation by preventing the disassembly of cancer cell epithelial junctional complex via the GSK-3ß/ß-catenin pathway.

Astragaloside IV protects new born rats from anesthesia-induced apoptosis in the developing brain.

Exposure to general anesthesia may cause severe neurotoxicity in developing brain due to neuronal apoptosis. Astragaloside IV (AS IV) has antioxidant and antiapoptosis properties; however, its effects on anesthesia-induced neuroapoptosis have not been studied. In the present study, we determined whether AS IV pre-treatment is able to reduce isoflurane exposure-induced neuroapoptosis in rats. New born rats were pre-treated with AS IV or solvent by oral gavage for three days, then exposed to isoflurane. The results showed that pre-treatment of AS IV significantly inhibited isoflurane-induced neural apoptosis in the hippocampus of new born rats, and such protection was accompanied by reduced levels of caspase-3, nuclear factor-κB activation and phosphorylated c-Jun N-terminal kinase, extracellular signal-regulated kinase and increased levels of B-cell lymphoma-2, glycogen synthase kinase-3ß, Klotho and phosphorylated protein kinase B. Furthermore, AS IV pre-treatment significantly alleviated isoflurane-induced oxidative stress and proinflammatory cytokine release in the rat hippocampus and serum. In summery, the results of the study demonstrated that AS IV is able to protect developing brain from anesthesia-induced neuroapoptosis via anti-oxidant and anti-inflammatory activities.

[Influence of Co-inhibiting mTORC2 and HSP90 on Proliferation Apoptosis of Multiple Myeloma Cells].

UNLABELLED: Objective：To explore the influence of co-inhibiting mTORC2 and HSP90 on the proliferation and apoptosis of multiple myeloma(MM) cell line U266. METHODS: During culture, the human MM cell line U266 were treated with 20 nmol/L of rapamycin, 600 nmol/L 17-AAG, 20 nmol/L of rapamycin + 600 nmol/L 17-AGG and phosphate-buffered saline (PBS), then the growth inhibition rate, morphologic changes, apoptosis rate and the expression of caspase 3 and ATK protein in U266 cells were compared and analyzed. RESULTS: The rapamycin and 17-AAG both could inhibit the growth of U266 cells, while the inhibitory effect of rapamycin in combination with 17-AAG on growth of U266 cells was significantly higher them that of rapamycin and 17-AAG alone and control (PBS); the apoptosis rate of U266 cells treated with rapamycin, 17-AAG and their combination was higher than that of control PBS groups, and the efficacy of 2 drug conbination was higher than that of control PBS group, and the efficacy of 2 drug combination was superior to single drug. The expression levels of caspase 3 and ATK in U266 cells treated with rapamycin, 17-AAG and their combination were higher and lower than those in control group respectively, and the efficacy of 2 drug combination was superior to signle drug. There were significant difference between them (P<0.05). CONCLUSION: The co-inhibition of mTORC2 and HSP90 can suppress the proliferation and induce the apoptosis of MM cells.

Secondary hemophagocytic lymphohistiocytosis induced by cholecystitis: A case report and a review of the literature.

BACKGROUND: Hemophagocytic lymphohistiocytosis (HLH) is a rare, life-threatening disease resulting from excessive activation and non-malignant proliferation of macrophages and T lymphocytes. Whether it can be caused by cholecystitis has not yet been reported in the world. CASE REPORT: A 4-year-old girl was admitted to hospital with cholecystitis. The patient was diagnosed with hemophagocytic lymphohistiocytosis after 3days of admission based on the results of laboratory tests showing hypofibrinogenemia, hypertriglyceridemia, thrombocytopenia, anemia and leukopenia. CONCLUSIONS: From this case experience, if a timely symptomatic treatment is given, the condition of the patient with secondary HLH can be alleviated. This is the first report of cholecystitis-induced hemophagocytic syndrome in the world also.

Therapeutic evaluation of monoclonal antibody-maytansinoid conjugate as a model of RON-targeted drug delivery for pancreatic cancer treatment.

Aberrant expression of the RON receptor tyrosine kinase, a member of the MET proto-oncogene family, contributes significantly to pancreatic cancer tumorigenesis and chemoresistance. Here we validate RON as a target for pancreatic cancer therapy using a novel anti-RON antibody Zt/g4-drug maytansinoid conjugates (Zt/g4-DM1) as a model for RON-targeted drug delivery to kill pancreatic cancer cells. In pancreatic cancer cell lines overexpressing RON, Zt/g4-DM1 rapidly induced receptor endocytosis, arrested cell cycle at G2/M phase, reduced cell viability, and subsequently caused massive cell death. These in vitro observations help to establish a correlation between the number of the cell surface RON receptors and the efficacy of Zt/g4-DM1 in reduction of cell viability. In mice, Zt/g4-DM1 pharmacokinetics in the linear dose range fitted into a two-compartment model with clearance in 0.21 ml/day/kg and terminal half-life at 6.05 days. These results helped to confirm a concentration-activity relationship for the BxPC-3 and other pancreatic cancer cell xenograft model with a tumoristatic dose at 3.02 mg/kg. Zt/g4-DM1 was effective in vivo against various xenograft PDAC growth but efficacy varied with individual cell lines. Combination of Zt/g4-DM1 with gemcitabine had a complete inhibition of xenograft pancreatic cancer growth. We conclude from these studies that increased RON expression in pancreatic cancer cells is a suitable targeting moiety for anti-RON ADC-directed drug delivery and anticancer therapy. Zt/g4-DM1 is highly effective alone or in combination with chemotherapeutics in inhibition of pancreatic cancer xenograft growth in preclinical models. These findings justify the use of humanized Zt/g4-DM1 for targeted pancreatic cancer therapy in the future.

Exosomes decrease sensitivity of breast cancer cells to adriamycin by delivering microRNAs.

While adriamycin (adr) offers improvement in survival for breast cancer (BCa) patients, unfortunately, drug resistance is almost inevitable. Mounting evidence suggests that exosomes act as a vehicle for genetic cargo and constantly shuttle biologically active molecules including microRNAs (miRNAs) between heterogeneous populations of tumor cells, engendering a resistance-promoting niche for cancer progression. Our recent study showed that exosomes from docetaxel-resistance BCa cells could modulate chemosensitivity by delivering miRNAs. Herein, we expand on our previous finding and explore the relevance of exosome-mediated miRNA delivery in resistance transmission of adr-resistant BCa sublines. We now demonstrated the selective packing of miRNAs within the exosomes (A/exo) derived from adr-resistant BCa cells. The highly expressed miRNAs in A/exo were significantly increased in recipient fluorescent sensitive cells (GFP-S) after A/exo incorporation. Gene ontology analysis of predicted targets showed that the top 30 most abundant miRNAs in A/exo were involved in crucial biological processes. Moreover, A/exo not only loaded miRNAs for its production and release but also carried miRNAs associated with Wnt signaling pathway. Furthermore, A/exo co-culture assays indicated that miRNA-containing A/exo was able to increase the overall resistance of GFP-S to adr exposure and regulate gene levels in GFP-S. Our results reinforce our earlier reports that adr-resistant BCa cells could manipulate a more deleterious microenvironment and transmit resistance capacity through altering gene expressions in sensitive cells by transferring specific miRNAs contained within exosomes.

Autoantibody response to Sui1 and its tissue-specific expression in hepatocellular carcinoma.

To investigate the immunogenicity of Homo sapiens putative translation initiation factor (Sui1) in hepatocellular carcinoma (HCC), enzyme-linked immunosorbent assay (ELISA) and Western blot were utilized to assess autoantibody responses to Sui1 in sera from HCC patients and healthy individuals. Indirect immunofluorescence (IIF) assay with cancer cells and immunohistochemistry (IHC) study with tissue array slides were performed to examine Sui1 expression profile in cancer cells and tissues. The data confirmed that the frequency of autoantibody to Sui1 in sera of HCC patients was 15.5 % (16/103), which was remarkably higher than that in sera of liver cirrhosis (LC) patients (3.3 %, 1/30), chronic hepatitis (CH) patients (0 %, 0/29), and normal human serum (NHS) (0 %, 0/82) (p < 0.01). IHC study showed that the Sui1 expression in HCC tissues was 26.7 % (16/60). The expression of Sui1 had the trend of increasing along with the cancer grades but no statistical significance (p > 0.05). In immunodiagnosis of HCC, the sensitivity and specificity of the anti-Sui1 antibody were 15.5 and 99.3 %, respectively. If both anti-Sui1 and alpha fetal protein (AFP) were simultaneously utilized as detective markers, 66.7 % (30/45) of HCC patients could be correctly distinguished. The results suggested that anti-Sui1 could be utilized as a supplementary serological marker for the detection of HCC and Sui1 might be associated to HCC carcinogenesis.