Screening and validation for core genes in osteoarthritic cartilage based on weighted gene co-expression network analysis.

OBJECTIVE: Osteoarthritis (OA) has the highest disability rate among chronic diseases. The burden on patients and public health care resources is increasingly evident due to increasing obesity rates and aging populations. So, there is still a lack of early diagnosis and treatment for OA. MATERIALS AND METHODS: A total of three OA cartilage tissue datasets (GSE1919, GSE32317, and GSE5235) were obtained from the Gene Expression Omnibus (GEO) database. Screening of differentially expressed genes and WGCNA of overlapping genes were performed using the R language package. Functional and immune infiltration analyses of overlapping genes were also carried out while hub genes were screened through LASSO regression analysis method and ROC curve. Finally, experimental validation was carried out through PCR and Western Blot analysis of rat cartilage. RESULTS: A total of 149 differentially expressed genes were screened, and they were mainly enriched in the cytokine-cytokine receptor interaction, rheumatoid arthritis, and interleukin (IL-17) signaling pathways. Four co-expression modules were obtained, of which the blue module was the most substantial morbidity associated with OA. Thirteen overlapping genes were identified based on significant module network topology analysis and differential genes, upon which their validation through LASSO regression analysis method and ROC curve was performed. From these, five signature genes were determined, before three potential core genes were finally identified after confirmation using the validation set. CONCLUSIONS: ATF3, FOSL2, and GADD45B may be hub genes to the osteochondropathy, and they are expected to be new biomarkers and drug targets in OA research.

Detection of CTCs and CSCs in the staging and metastasis of non-small cell lung cancer based on microfluidic chip and the diagnostic significance.

OBJECTIVE: Dynamic monitoring of CTCs/CSCs can assist in the diagnosis and prognosis of tumors. This study explores the diagnostic significance of microfluidic chip technology in the detection of CTCs/CSCs in clinical staging and metastasis of patients with non-small cell lung cancer (NSCLC). That lays a solid foundation for the use of microfluidic chips to monitor CTCs/CSCs for the stage and metastasis of patients with non-small cell lung cancer. PATIENTS AND METHODS: This study collected 80 patients with lung cancer from October 2017 to October 2018. Meanwhile, 30 healthy people and 30 patients with benign lung diseases were selected during the same period as the control group 1 and the control group 2, respectively. CellSearch (Huntington Valley, PA, USA) and microfluidic chip were used to detect CTCs, the sensitivities were recorded. ELISA methods were used to detect the concentrations of tumor markers VEGF-C, CEA, and CA125 in serum, and their association with CTCs and CSCs was analyzed. In addition, after 3 months, we followed up 40 patients with lung cancer, recorded their prognosis, and extracted peripheral blood to detect changes in their CTCs and CSCs. The CellSearch (Huntington Valley, PA, USA) system and the microfluidic chip system were used to detect the CTCs in patients with lung cancer, and the sensitivity and specificity of the patients were analyzed. The changes in CTCs and CSCs in the peripheral blood of the patient were recorded. RESULTS: It can be seen that the positive rate of CTCs and CSCs is not significantly correlated with the patients' age, gender, pathological type (adenocarcinoma, squamous cell carcinoma), etc. They are significantly correlated with clinical stage (I + II and III + IV) and metastasis (metastasis and non-metastasis) (p<0.01). Then, we divided the patients into groups for testing, and analyzed the association between different groups of patients and CTCs and CSCs. Compared with control group 1 and control group 2, the positive rates of CTCs and CSCs in lung cancer metastasis group and non-metastasis group were significantly different (p<0.05). Compared with the control group 1 and control group 2, the positive rates of CTCs and CSCs in stage I + II and III + IV of lung cancer were significantly different (p<0.05). The positive rate was significantly higher in the cancer metastasis group (p<0.05). The concentrations of tumor markers VEGF-C, CEA, CA125 in the serum of patients were consistent with CTCs-negative and CTC-positive lung cancer, with significant differences (p<0.05). CSCs negative and CSCs positive patients have similar results. Subsequently, we analyzed the sensitivity and specificity of CSCs, CTCs, and tumor markers for the diagnosis of NSCLC. The results showed that the sensitivity of CSCs and CTCs to diagnose patients was significantly higher than that of tumor markers. CONCLUSIONS: This study shows that our microfluidic chip device can exhibit relatively good performance and can better detect CTCs and CSCs. Monitoring CTCs and CSCs of patients can provide a basis for judging the stage and metastasis of patients.

Utilizing integrating network pharmacological approaches to investigate the potential mechanism of Ma Xing Shi Gan Decoction in treating COVID-19.

Beginning in December 2019, coronavirus disease 2019 (COVID-19), due to 2019-nCoV infection, emerged in Wuhan and spread rapidly throughout China and even worldwide. Employing combined therapy of modern medicine and traditional Chinese medicine has been proposed, in which Ma Xing Shi Gan Decoction (MXSGD) was recommended as a basic prescription and applied widely in the clinical treatment of COVID-19. We investigated the underlying mechanism of MXSGD in treating COVID-19 utilizing the approaches of integrating network pharmacology. A total of 97 active ingredients of MXSGD were screened out, and 169 targets were predicted. The protein-protein interaction network exhibited hub targets of MXSGD, such as Heat shock protein 90, RAC-alpha serine/threonine-protein kinase, Transcription factor AP-1, Mitogen-activated protein kinase 1, Cellular tumor antigen p53, Vascular endothelial growth factor A, and Tumour necrosis factor. Gene Ontology functional enrichment analysis demonstrated that the biological processes altered within the body after taking MXSGD were closely related to the regulation of such processes as the acute inflammatory response, chemokine production, vascular permeability, response to oxygen radicals, oxidative stress-induced apoptosis, T cell differentiation involved in the immune response, immunoglobulin secretion, and extracellular matrix disassembly. KEGG enrichment analysis indicated that the targets of MXSGD were significantly enriched in inflammation-related pathways, immunomodulation-related pathways, and viral infection-related pathways. The therapeutic mechanisms of MXSGD on COVID-19 may primarily involve the following effects: reducing inflammation, suppressing cytokine storm, protecting the pulmonary alveolar-capillary barrier, alleviating pulmonary edema, regulating the immune response, and decreasing fever.

CircRNA_100876 promote proliferation and metastasis of breast cancer cells through adsorbing microRNA-361-3p in a sponge form.

OBJECTIVE: This study was designed to investigate the expression level of circRNA_100876 in breast cancer (BC) tissues or cells, and to further explore whether it can promote cell metastasis and proliferative capacity via targeting microRNA- 361-3 p. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine the expression of circRNA_100876 in 50 pairs of BC tissue specimens and corresponding adjacent ones, and the correlation between circRNA_100876 expression and prognosis of patients with BC was analyzed. Meanwhile, qRT-PCR was further performed to verify circRNA_100876 level in BC cell lines. In addition, circRNA_100876 knockdown model was constructed using lentivirus and transfected in BC cells. Subsequently, the impact of circRNA_100876 on BC cell function was analyzed using Cell Counting Kit-8 (CCK-8), transwell and clone formation assays. The interplay between circRNA_100876 and microRNA- 361-3 p was verified using the Luciferase reporter gene assay and cell reverse experiment. RESULTS: QRT-PCR results showed that circRNA_100876 level in BC tissues was conspicuously higher than that in the adjacent tissues, and the patients with distant metastasis had higher expression than those without. Moreover, patients with a high expression of circRNA_100876 had a relatively lower overall survival rate. Compared with the NC group, the cell proliferation and invasion ability of circRNA_100876 knockdown group was conspicuously decreased. QRT-PCR revealed that microRNA-361-3p and circRNA_100876 showed a negative correlation in the expression level of genes in BC tissues. In addition, the results of the Luciferase reporter gene assay confirmed that circRNA_100876 can be targeted by microRNA-361-3p through their binding site. CONCLUSIONS: High expression of circRNA_100876 is conspicuously positively relevant to poor prognosis of BC patients. Additionally, circRNA_100876 is able to promote BC metastasis as well as proliferative capacity by modulating microRNA-361-3p expression.

Up-regulation of miRNA-105 inhibits the progression of gastric carcinoma by directly targeting SOX9.

OBJECTIVE: MicroRNAs (miRNAs) are involved in the tumorigenesis and progression of multiple tumor types and function as either tumor suppressor genes or oncogenes. This study was designed to investigate the functional behaviors and regulatory mechanisms of miR-105 in the progression of gastric carcinoma. PATIENTS AND METHODS: 24 pairs of patients with gastric carcinoma were enrolled in this study. The levels of miR-105 in gastric carcinoma tissues and cells were determined using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay. The biological functions of miR-105 in gastric carcinoma cell were detected by colony formation, transwell invasion and wound-healing assay. Luciferase activity assay and immunoblotting assay were applied to validate the direct target of miR-105. The expression of SRY-Box 9 (SOX9) was detected using immunofluorescence staining assay. Furthermore, the role of miR-105 on the growth of gastric carcinoma cell was examined in the established xenograft model. The role of miR-105 in the metastasis of gastric carcinoma cell in vivo, an experimental metastasis assay was performed. RESULTS: Herein, we proved that miR-105 was down-regulated in gastric carcinoma specimens as well as gastric cancer cells. Up-regulation of miR-105 suppressed the colony formation and aggressiveness traits of gastric carcinoma cell lines BGC823 and SGC7901 in vitro. Furthermore, over-expression of miR-105 inhibited the tumor growth as well as lung metastasis of gastric carcinoma cell in vivo. Further investigation identified SOX9 was the target gene of miR-105 in gastric cancer and its expression was negatively associated with the expression of miR-105 in gastric carcinoma tissues. Finally, overexpression of SOX9 partially reversed the influence of miR-105 on the growth and aggressiveness of gastric carcinoma cell. CONCLUSIONS: These results revealed the crucial role of miR-105 in the progression and metastasis of gastric carcinoma, which indicated the potential application of miR-105 in the treatment of gastric carcinoma.

MiR-126 induces myeloma cell line Karpas707 apoptosis by downregulating anti-apoptotic protein MCL.

OBJECTIVE: Myeloma seriously threats human life and health and needs more efficacy treatment method in the clinic. MiR-126 regulates cell proliferation and apoptosis. This study explores the regulatory role of miR-126 in myeloma and related molecular mechanism. MATERIALS AND METHODS: MiR-126 and control were synthesized and transfected to myeloma cell line Karpas707 using Lipofectamine. Cell apoptosis was evaluated by MTT assay, caspase-3 activity detection, and flow cytometry. Myeloid cell leukemin (MCL) siRNA and plasmid were transfected to Karpas707 cells to test its impact on cell apoptosis. RESULTS: MTT assay revealed that miR-126 significantly restrained Karpas707 cell growth (p=0.0017). Cell apoptosis detection showed that miR-126 significantly promoted phosphatidylserine eversion and caspase-3 activation (p=0.031), and downregulated MCL level (p=0.017). MCL siRNA markedly enhanced Karpas707 cell apoptosis induced by miR-126 (p=0.024), while the MCL overexpression apparently inhibited Karpas707 cell apoptosis induced by miR-126 (p=0.0073). CONCLUSIONS: MiR-126 induces Karpas707 cell apoptosis by downregulating anti-apoptotic protein MCL, which provides a theoretical basis for the target selection of myeloma.

The clinical effects of gonadotropin-releasing hormone agonists for the treatment of children patients with central precocious puberty.

OBJECTIVE: We explored the clinical effect of gonadotropin-releasing hormone agonists for the treatment of children patients with central precocious puberty. PATIENTS AND METHODS: From March 2012 to October 2015, 100 cases of children patients with central precocious puberty were enrolled in this study. Intramuscular injection of acetic acid triptorelin (50 to 100 pLg/kg) was made once every 4 weeks, and the treatment lasted for 4 months. Patients' bone age/height differentials (DBA/DCA), bone age (BA), growth velocity (GV) and predicted adult height (PAH) were determined before and after treatment (after 6, 12, 24, 36 months). Differences before and after treatment were analyzed. DBA/DCA, BA and PAH values 6, 12, 24, 36 months after treatment were significantly different compared with those before treatment. RESULTS: Sexual development symptoms in children patients were significantly improved 4 months after treatment (p<0.05). All patients completed the treatment, without any adverse drug reaction or severe complication. After one course of treatment (4 months), patients' uterus and ovarian volumes shrank, FSH level peaked, and LH level was reduced, compared to those before treatment. CONCLUSIONS: Acetate acid triptorelin is safe and reliable for treating central precocious puberty. We achieved the excellent clinical curative effect and were able to delay the growth rate in children patients. The predicted height and final height were improved.

Overexpression of Ecm22 improves ergosterol biosynthesis in Saccharomyces cerevisiae.

Ergosterol biosynthesis in Saccharomyces cerevisiae is complex and the underlying mechanism of regulation remains unclear. To clarify the influence of transcriptional regulation on the ergosterol content, transcription factor Ecm22 was overexpressed in S. cerevisiae. Results showed that the overexpression of ECM22 led to an increased invasive growth. Fluconazole susceptibility testing indicated that strains overexpressing ECM22 could grow at 20 µg(fluconazole)  ml-1 . By contrast, the control failed to grow at 16 µg(fluconazole)  ml-1 . Among truncated ECM22 fragments, only the 1440-bp DNA fragment exerted almost the same impact on ergosterol content as that of the full-length gene. In a 5-l bioreactor, the highest ergosterol yield of the recombinant reached 32∙7 mg g(dry cell weight) -1 , which was increased by about 20% compared with that of the control. In this work, a novel approach for enhancing the ergosterol production by overexpressing a transcription factor in S. cerevisiae was developed.

Differentiation between malignant and benign breast masses: combination of semi-quantitative analysis on DCE-MRI and histogram analysis of ADC maps.

AIM: To investigate the performance of combined semi-quantitative analysis on dynamic contrast enhanced (DCE) magnetic resonance imaging (MRI) and histogram analysis of diffusion-weighted imaging (DWI) for distinguishing malignant from benign breast masses. MATERIALS AND METHODS: This study included 178 patients with breast masses (benign:malignant=88:9) who underwent both DCE-MRI and DWI. The semi-quantitative parameters, derived from DCE-MRI, included maximum slope of increase (MSI), signal intensity slope (SIslope), initial percentage of enhancement (Einitial), percentage of peak enhancement (Epeak), early signal enhancement ratio (ESER), and second enhancement percentage (SEP). Histogram parameters derived from apparent diffusion coefficient (ADC) maps included ADCmin, ADCmax, ADCmean, ADC10, ADC25, ADC50, ADC75, ADC90, skewness, and kurtosis. All parameters were compared between malignant and benign groups, and their differences were tested using independent-samples t-test or Mann-Whitney test. Receiver operating characteristic (ROC) curves were used to determine the diagnostic value of each significant parameter. RESULTS: Among semi-quantitative parameters, SIslope exhibited the best diagnostic performance in predicting malignancy (cut-off value, 0.096; ROC, 0.756; sensitivity, 86.7%; specificity, 61.4%). Among histogram parameters, ADC10 exhibited the best diagnostic performance in predicting malignancy (cut-off value, 1.051; ROC, 0.885; sensitivity, 86.7%; specificity, 84.1%). The optimal diagnostic performance of combined ADC10 and SIslope (area under curve [AUC], 0.888; sensitivity, 82.2%; specificity, 95.5%) was significantly better than SIslope alone (p<0.001). Moreover, the combination showed higher AUC (0.888 versus 0.885) than ADC10 alone, but the difference was not statistically significant (p=0.914). CONCLUSION: SIslope and ADC10 are significant predictors for breast malignancy. The combination of DCE-MRI and DWI improves differentiating performance.

The effects of add-on exenatide to insulin on glycemic variability and hypoglycemia in patients with type 1 diabetes mellitus.

OBJECTIVE: To investigate the effect of add-on exenatide to insulin on glycemic excursion and the counter-regulatory hormone in response to hypoglycemia in patients with type 1 diabetes mellitus (T1DM). METHODS: 30 patients with T1DM were recruited and randomly assigned to exenatide + insulin-treated group (group 1, n = 15) or insulin-only-treated group (group 2, n = 15) for 4 weeks. All patients had continuous glucose monitor system (CGMS) applied at before (week-0) and after (week-4) treatment to evaluate the glycemic variability. All patients had an arginine-stimulated test at before and after treatment. Six patients from each group also had hypoglycemic clamp test to assess counter-regulatory hormone level. RESULTS: Patients in the exenatide group had significant reductions in body weight, body mass index (BMI), total insulin dose, bolus insulin dose, fructosamine, and glycemic excursion after 4 weeks' treatment. Compared with patients in group 2, the mean amplitude of glycemic excursion (MAGE) and coefficient of variation (CV) of exenatide group decreased significantly. Similarly, a significant decrease of glucagon (GLC) in the arginine-stimulated test was found in group 1. No significant changes of GLC, growth hormone (GH), cortisol (COR), epinephrine (E), and norepinephrine (NE) were found in both groups during hypoglycemia clamp test. However, patients who had residual islet function in group 1 showed an upward trend of basic C-peptide (C-P) and GLC during the hypoglycemia period. CONCLUSION: Although exenatide could inhibit glucagon secretion during euglycemia or hyperglycemia in patients with T1DM, it has no effect on GLC and counter-regulatory hormones during hypoglycemia clamp in patients with no functional residual islet test.

Role of local allergic inflammation and Staphylococcus aureus enterotoxins in Chinese patients with chronic rhinosinusitis with nasal polyps.

OBJECTIVE: To investigate the role of local allergic inflammation and Staphylococcus aureus enterotoxins in chronic rhinosinusitis with nasal polyps. METHODS: This study included 36 patients with chronic rhinosinusitis with nasal polyps and 18 controls. Total immunoglobulin E, eosinophil cationic protein, staphylococcal enterotoxin types A and B specific immunoglobulin E, staphylococcal enterotoxin types A and B, and myeloperoxidase levels were determined. RESULTS: Four patients with chronic rhinosinusitis with nasal polyps had a local allergy. All chronic rhinosinusitis with nasal polyps patients tested negative for staphylococcal enterotoxin types A and B specific immunoglobulin E. The chronic rhinosinusitis with nasal polyps group had significantly elevated staphylococcal enterotoxin types A and B levels in the supernatant. Fourteen patients belonged to the eosinophilic chronic rhinosinusitis with nasal polyps group and the others were characterised as having non-eosinophilic chronic rhinosinusitis with nasal polyps. CONCLUSION: Local allergy may play a role in chronic rhinosinusitis with nasal polyps, independent of staphylococcal enterotoxin superantigens. Staphylococcal enterotoxins may be important in the pathogenesis of chronic rhinosinusitis with nasal polyps; however, their roles as superantigens were not confirmed in this study. In Chinese subjects, chronic rhinosinusitis with nasal polyps usually manifests as a neutrophilic inflammation.

Oxymatrine suppresses proliferation and induces apoptosis of hemangioma cells through inhibition of HIF-1a signaling.

Oxymatrine (OMT), a natural quinolizidine alkaloid, has been known to have anti-inflammation, anti-anaphylaxis, and chemopreventive effects on various cancer cells. To clarify the underlying role and molecular mechanisms of OMT in human hemangioma (HA), in the present study, we examined the expression of hypoxia-inducible factor-1a (HIF-1a) and vascular endothelial growth factor (VEGF) in different phases of human HA. After HA derived endothelial cells (HDEC) were pretreated with different concentrations of OMT, cell proliferation, apoptosis, and cycle distribution were evaluated by MTT assay and flow cytometry analysis, respectively. The effects of OMT on expression of HIF-1a signaling were determined by real-time PCR and western blot assays. Our results showed that, the expression of HIF-1a and VEGF was significantly increased in proliferating phase HA, but decreased in involuting phase HA. Moreover, OMT in vitro inhibited proliferative activities and induced cell apoptosis and cycle arrest in G0/G1 phase in HA cells with decreased expression of HIF-1a, VEGF, Bcl-2, and CyclinD1, and increased expression of p53. Taken together, our findings suggest that, the expression of HIF-1a and VEGF is increased in proliferating phase HA, and OMT suppresses cell proliferation and induces cell apoptosis and cycle arrest in proliferative phase HA through inhibition of the HIF-1a signaling pathway, suggesting OMT may provide a novel therapeutic strategy for the treatment of HA.

Exploring the osteoarthritis-related genes by gene expression analysis.

OBJECTIVE: Osteoarthritis (OA) is a most common chronic degenerative joint lesion, which affects both cartilage and bone. A better understanding of the gene expression profiling of OA may help understanding the pathogenesis of OA and finding the therapy targets for OA treatment. MATERIALS AND METHODS: GSE8077 was downloaded from Gene Expression Omnibus (GEO) including 5 OA rats induced by anterior cruciate ligament transection and partial medial meniscectomy and 5 rats that were performed sham surgery as control. Differentially expressed genes (DEGs) between OA group and control group were identified by t-test with p < 0.05 and the coding genes that transcription factors corresponded were screened by TRANSFAC. Then Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis for DEGs and transcription factors were performed. The DEGs and transcription factors were integrated with information from STRING database to construct PPI network. RESULTS: A total of 119 up-regulated genes, 39 down-regulated genes and 9 transcription factors were identified in OA sample. The GO enrichment analysis showed that 119 up-regulated genes were significantly enriched in blood vessel development and KEGG pathway enrichment showed that genes were involved in circadian rhythm pathway. In the PPI network, Cd44, Mmp13, Timp1 and Igf1 showed higher degrees. CONCLUSIONS: The screened genes could provide a new and comprehensive view for treatment of OA.

Pathway enrichment analysis of human osteosarcoma U-2 OS bone cells expose to dexamethasone.

OBJECTIVE: Osteosarcoma is the second highest cause of cancer-related death in children, mainly due to development of often fatal metastasis, usually in the lungs. Glucocorticoids play an important role in the treatment of a number of inflammatory diseases and immune diseases. The objective of this study was to explore the molecular mechanism of osteosarcoma in response to dexamethasone (DEX, a kind of synthetic glucocorticoid), with a view to obtain information on the pathways activated by DEX. MATERIALS AND METHODS: By using the GSE6711 Affymetrix microarray data accessible from Gene Expression Omnibus database, we first identified the differentially expressed genes (DEGs) among different time course treatment with dexamethasone of each isoform, and the DEGs among cells expressing different GR isoforms, followed by the pathway enrichment analysis of the DEGs. RESULTS: The results indicated that DEX could inhibit osteosarcoma cell proliferation and promote osteosarcoma cell apoptosis through induction of lots of related genes expression at the transcription level. CONCLUSIONS: Our data provide a comprehensive bioinformatics analysis of pathways which may be involved in the response to glucocorticoids.

Novel Y-chromosome polymorphisms in Chinese domestic yak.

Y-chromosome-specific haplotypes (Y-haplotypes) constructed using single nucleotide polymorphisms (Y-SNPs) in the MSY (male-specific region of the Y-chromosome) are valuable in population genetic studies. But sequence variants in the yak MSY region have been poorly characterized so far. In this study, we screened a total of 16 Y-chromosome-specific gene segments from the ZFY, SRY, UTY, USP9Y, AMELY and OFD1Y genes to identify Y-SNPs in domestic yaks. Six novel Y-SNPs distributed in the USP9Y (g.223C>T), UTY19 (g.158A>C and g.169C>T), AMELY2 (g.261C>T), OFD1Y9 (g.165A>G) and SRY4 (g.104G>A) loci, which can define three Y-haplotypes (YH1, YH2 and YH3) in yaks, were discovered. YH1 was the dominant and presumably most ancient haplotype based on the comparison of UTY19 locus with other bovid species. Interestingly, we found informative UTY19 markers (g.158A>C and g.169C>T) that can effectively distinguish the three yak Y-haplotypes. The nucleotide diversity was 1.7 × 10(-4) ± 0.3 × 10(-4) , indicating rich Y-chromosome diversity in yaks. We identified two highly divergent lineages (YH1 and YH2 vs. YH3) that share similar frequencies (YH1 + YH2: 0.82-0.89, YH3: 0.11-0.18) among all three populations. In agreement with previous mtDNA studies, we supported the hypothesis that the two highly divergent lineages (YH1 and YH2 vs. YH3) derived from a single gene pool, which can be explained by the reunion of at least two paternal populations with the divergent lineages already accumulated before domestication. We estimated a divergence time of 408 110 years between the two divergent lineages, which is consistent with the data from mitochondrial DNA in yaks.

Neonatal sevoflurane anesthesia induces long-term memory impairment and decreases hippocampal PSD-95 expression without neuronal loss.

AIM: Volatile anesthetics are widely used in the clinic, and sevoflurane is the most prevalent volatile anesthetic in pediatric anesthesia. Recent findings question the potential risks of volatile anesthetics on brain development. Evidence suggests that sevoflurane may cause neuronal deficiency. This study investigates the long-term effect of sevoflurane in the developing brain. MATERIALS AND METHODS: We anesthetized 7 day-old rats for 4 h with 2.5% sevoflurane. A Morris water maze was used to evaluate hippocampal function 7 weeks after sevoflurane exposure. Nissl staining was performed to analyze neuronal loss. PSD-95 (postsynaptic density protein-95) expression in the hippocampus was measured using a western blot. RESULTS: The exposure to 2.5% sevoflurane caused long-term deficits in hippocampal function and decreased hippocampal PSD-95 expression without neuronal loss. This study demonstrates that P7 rats exposed for 4 h to 2.5% sevoflurane have significant spatial learning and memory impairment 7 weeks after anesthesia. In addition, PSD-95 expression in the hippocampus decreased at P56 without neuronal loss. CONCLUSIONS: These data suggest that sevoflurane causes neurotoxicity in the developing brain, which may be attributed to decreased PSD-95 in the hippocampus.

Role of stromal-derived factor-1/CXCR4 in neo-intimal repair.

Neo-intimal hyperplasia is one of the major causes of restenosis in which stromal cell-derived factor-1 (SDF-1α) and its receptor CXCR4 play an important role. In a rat common carotid artery balloon injury model, the number of CD34(+)CXCR4(+) cells was significantly increased immediately after injury (p < 0.01), followed by a gradual decrease to baseline seven days after the injury. Furthermore, the plasma (SDF-1α) level was markedly elevated, and peaked 24 hours after injury (p < 0.01), followed by a rapid decrease to baseline level seven days after the injury. In the injured common carotid artery, the mRNA expression of (SDF-1α) was elevated immediately after injury, followed by a gradual decline, but that of CXCR4 was increased four days after injury. Immuno-histochemistry displayed CXCR4-positive staining one day after injury, which then gradually increased and continued for at least one month. In addition, administration of AMD3100 (200 ng/kg, i.p.), a CXCR4 antagonist, did not affect the number of CD34(+)CXCR4(+) cells, the elevated level of plasma (SDF-1α) and expression of (SDF-1α) mRNA. The expression of CXCR4 mRNA and protein however was markedly decreased, and detectable CXCR4-positive cells occurred four days after injury, followed by a decreased intensity of staining. We also found that, three months after balloon injury, stenosis of the carotid artery intima in the group that received AMD3100 was significantly less than in the untreated group (p < 0.05). Therefore, (SDF-1α)/CXCR4 played a crucial role in the intimal hyperplasia, and restenosis may have be attenuated after inhibition of CD34(+)CXCR4(+) cells in the intima.

Differential diagnosis of inflammatory myofibroblastic tumour and low-grade myofibroblastic sarcoma: two case reports with a literature review.

Inflammatory myofibroblastic tumour (IMT) and low-grade myofibroblastic sarcoma (LGMS) have similar morpho logical and immunophenotypic features, but LGMS is more malignant than IMT and the treatment requires a wider surgical margin plus post-operative chemotherapy or radiotherapy. To date, only 28 cases of IMT and two cases of LGMS have been reported in the laryngopharynx. Recent studies have suggested that anaplastic lymphoma kinase (ALK) and cytokeratin are important markers for differentiating between the two tumours. Here, two cases involving different myofibroblastic tumours of the larynx are reported. Based on the histological and immunohistochemical results, case 1 was diagnosed as IMT involving the right arytenoepiglottic fold, while case 2 was diagnosed as LGMS involving the epiglottic-glossal surface. There was no recurrence or metastasis in either case after post-operative follow-up (12 and 14 months, respectively). It is difficult to distinguish IMT from LGMS; both morphological and immunohistological analyses are required.

Angiopoietin-2 inhibits the growth of tongue carcinoma without affecting expression of vascular endothelial growth factor.

Angiopoietin-2 (Ang-2) has been identified as an important factor in tumour angiogenesis through its action in blocking the stabilizing actions of Ang-2 and leading to new tumour vessel growth in the presence of vascular endothelial growth factor (VEGF). In the authors' previous study, over-expression of Ang-2 in Tca8113 tongue tumour cells inhibited growth. The current study aims to clarify the mechanisms of Ang-2-mediated tumour growth inhibition and its role in the regulation of VEGF expression. These studies used tumours derived from Ang-2-transfected Tca8113 cells injected into nude mice. The results showed that Ang-2-transfected tumours displayed aberrant angiogenic vessels with few associated smooth muscle cells. No detectable differences in VEGF expression were observed between Ang-2-transfected and parental tumours. Tumours produced by the Ang-2 transfection also had a higher apoptosis index and lower tumour cell proliferative index than tumours in the control groups. These observations suggest that enhanced expression of Ang-2 has no effect on VEGF expression and results in tumour vessel regression and inhibition of tumour growth.

Virtual screening for finding natural inhibitor against cathepsin-L for SARS therapy.

Recently Simmons et al. reported a new mechanism for SARS virus entry into target cells, where MDL28170 was identified as an efficient inhibitor of CTSL-meditated substrate cleavage with IC(50) of 2.5 nmol/l. Based on the molecule fingerprint searching method, 11 natural molecules were found in the Traditional Chinese Medicines Database (TCMD). Molecular simulation indicates that the MOL376 (a compound derived from a Chinese medicine herb with the therapeutic efficacy on the human body such as relieving cough, removing the phlegm, and relieving asthma) has not only the highest binding energy with the receptor but also the good match in geometric conformation. It was observed through docking studies that the van der Waals interactions made substantial contributions to the affinity, and that the receptor active pocket was too large for MDL21870 but more suitable for MOL736. Accordingly, MOL736 might possibly become a promising lead compound for CTSL inhibition for SARS therapy.