Application of personalized spacers made with 3D-printed mold in the two-stage procedure for periarticular infection.

BACKGROUND: Cement spacers treat periarticular infection after bone tumor resection in patients with bone defects. Complications such as poor joint function, poor soft tissue reconstruction, and poor postoperative daily living ability are present. We present a case of periarticular infection treated successfully after distal femoral osteosarcoma surgery with a personalized spacer made with a 3D-printed mold. CASE REPORT: A two-stage procedure was performed on an 18-year-old patient with high-grade conventional osteosarcoma of the left distal femur. After two biopsies, the boy developed a periarticular infection of the affected limb during neoadjuvant chemotherapy. We had a microbiologically confirmed methicillin-resistant Staphylococcus aureus (MRSA) infection. Because of the infection risk associated with primary joint replacement, a two-stage procedure was planned. In the first stage of surgery, we prepared a personalized spacer using a 3D-printed mold, antibiotic-loaded polymethylmethacrylate (PMMA), and an intramedullary needle. This spacer restored the function of the knee joint and the daily activities of the affected limb, and the infection was effectively eradicated. This spacer was firmly fixed two years after the surgery, and there were no surgical or spacer-related complications. The patient underwent a second stage of surgery to replace a permanent metal mega-prosthesis, and the knee joint functions returned to near normal. CONCLUSIONS: This case report describes limb-salvage surgery following distal femoral resection for periarticular infection. The personalized spacers prepared by a 3D-printed mold can be used in periarticular infection after long bone resection, mega-prosthetic infection, or limb-salvage surgery for temporary joints in small children.

Clinical efficacy of modified sacral fixation under Leonardo da Vinci robot laparoscopy for pelvic organ prolapse.

OBJECTIVE: The aim of this study was to analyze the clinical efficacy of modified sacral fixation under Leonardo da Vinci robot laparoscopy for pelvic organ prolapse (POP). PATIENTS AND METHODS: Sixty POP patients admitted to our hospital from January 2020 to December 2021 were picked and divided into Group A (laparoscopic Y-mesh, n = 20), Group B (laparoscopic sacrovaginal fixation, n = 20), and Group C (da Vinci robotic sacral fixation, n = 20). These three groups were compared in terms of the perioperative indexes, such as operation time, intraoperative blood loss, postoperative indwelling catheter days, anal exhaust time, postoperative hospitalization days, etc. The occurrence of short-term and long-term complications in the three groups was compared. The changes of the following index values in the POP quantification system (POP -Q) staging before and 1 year after surgery were recorded and compared among the three groups. It mainly includes the midline of the anterior vaginal wall at 3 cm from the hymenal margin (Aa), the farthest point of the anterior vaginal vault from point Aa (Ba), the farthest point of the ectocervix (C), the location of the posterior vaginal vault or rectal uterine trap (D), the midline of the posterior vaginal wall at 3 cm from the hymenal margin (Ap), and the reflection of the posterior vaginal vault at the farthest point from the Ap point (Bp) values. The changes in Pelvic Floor Distress Inventory-Short Form 20 (PFDI-20) and Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12) were recorded and compared before and 1 year after the operation. RESULTS: The patients in Group C had significantly lower intraoperative bleeding, postoperative indwelling catheter days, anal exhaust time, and postoperative hospitalization days compared with those in Group A and Group B (p < 0.05). There existed no statistical difference in the incidence of short-term and long-term complications between Group B and Group C (p > 0.05), but both were much lower than Group A (p < 0.05). The differences in POP-Q staging, PFDI-20 scale, and PISQ-12 scale were not statistically significant among the three groups before surgery (p > 0.05), and the POP-Q staging Aa, Ba, C, D, Ap, and Bp values, PFDI-20 scale, and PISQ-12 scale were strongly improved in three groups after the surgery (p < 0.05). However, the POP-Q staging, PFDI-20 scale, and PISQ-12 scale among the three groups had no obvious difference after the surgery (p > 0.05). CONCLUSIONS: The efficacy of modified sacral fixation under Leonardo da Vinci robot laparoscopy for POP was comparable to that of laparoscopic Y-mesh treatment and laparoscopic sacral vaginal fixation. However, da Vinci's robotic sacral fixation had the advantages of less intraoperative bleeding and faster postoperative recovery, which helped patients recover quickly and improved their quality of life.

Correlations of ultrasound and pathological features of thyroid carcinoma with TC-1 mRNA and protein expression.

OBJECTIVE: To investigate the correlations of ultrasound and pathological characteristics of thyroid carcinoma through evaluating the messenger ribonucleic acid (mRNA) level and protein expression of thyroid cancer-1 (TC-1). PATIENTS AND METHODS: The patients with papillary thyroid carcinoma (PTC) hospitalized in our hospital were enrolled. Then, real-time fluorescence quantitative polymerase chain reaction (qPCR) and immunohistochemistry (IHC) streptavidin-peroxidase (SP) technique were applied to measure the mRNA and protein expression levels of TC-1 in PTC and corresponding adjacent tissues (NCE) of 50 patients. The relations with clinicopathological and ultrasound characteristics were analyzed. RESULTS: The expression of TC-1 mRNA in PTC tissues was statistically higher than that in corresponding adjacent tissues and significantly correlated with tumor-node-metastasis (TNM) stage, pathological grade, and lymph node metastasis of PTC (p<0.05). According to IHC, TC-1 positive expression was mainly found in the cytoplasm in PTC samples, which was statistically increased compared to adjacent tissues (p<0.05). Western blotting results revealed that the relative protein expression of TC-1 in PTC tissues was 2.646±195, which was significantly higher than that in corresponding adjacent tissues (892±76) (p<0.05). The TC-1 protein expression also showed significant associations with TNM stage, pathological grade, and lymph node metastasis of patients (p<0.05). The level of TC-1 mRNA in PTC tissues with micro-calcification detected by ultrasound (87.46±49.55) was higher than that in those without micro-calcification (38.46±29.15) (p<0.05). CONCLUSIONS: The expression of TC-1 plays an important role in the occurrence and development of PTC. Ultrasound characteristics reflect the expression of TC-1 in PTC tissues to some extent, providing a certain value in evaluating the prognosis of PTC.

Role and mechanism of Dvl3 in the esophageal squamous cell carcinoma.

OBJECTIVE: The purpose of this project was to investigate the expression of Dishevelled-3 (Dvl3) in esophageal squamous cell carcinoma and cultured cells, and to determine the consequence of Dvl3 silencing in the tumorous properties of esophageal squamous cell carcinoma cells. PATIENTS AND METHODS: The expression of Dvl3 mRNA and protein in 50 cases of esophageal squamous cell carcinoma was detected. The expression of Dvl3 mRNA and protein was significantly elevated in esophageal squamous cell carcinoma tissues compared with atypical hyperplasia and normal esophageal mucosa. RESULTS: Dvl3 promoted the proliferation of esophageal squamous cell carcinoma cells and cell migration of cells expressing Dvl3 siRNA was significantly lower than that of the non-transfected cells. Flow cytometry showed that silencing Dvl3 promoted apoptosis of esophageal squamous cell carcinoma. Dvl3 overexpression cells in the subcutaneous tissue of nude mice promoted the formation of tumors. The expression of Dvl3 was associated with invasion and metastasis of the esophageal squamous cell carcinoma. CONCLUSIONS: Overall, down-regulation of Dvl3 expression can control the progression of esophageal squamous cell carcinoma, inhibit the growth and promote the apoptosis of tumor cells. Thus, Dvl3 has potential applications for early diagnosis, prognosis and therapeutics in the esophageal squamous cell carcinoma.

Expression and function of peripheral blood miRNA16a in patients with ankylosing spondylitis.

OBJECTIVE: Ankylosing spondylitis (AS) is a progressive spinal disease presented as rheumatoid factor negative. As an autoimmune disorder, AS is featured with an inflammatory change of tendon and ligament, accompanied by elevates serum levels of inflammatory factors. MicroRNA (miR) participates in the regulation of various diseases including tumor, inflammation, cardiovascular disease and immune response. MiR16a exerts critical roles in inflammatory disease. Its function in AS, however, has not been fully illustrated. PATIENTS AND METHODS: AS patients (at stable and active phase) and healthy controlled individuals were recruited to test peripheral expression of miR16a by Real-time PCR (RT-PCR). Enzyme-linked immunosorbent assay (ELISA) was used to test serum helper T cell 1 (Th1) cytokine levels including interferon (IFN)-γ, tumor necrosis factor-α (TNF-α) and Th2 cytokines including interleukin-4 (IL-4) and IL-10. The correlations between miR16a and cytokine levels, C reactive protein (CRP), erythrocyte sedimentation rate (ESR) and AS activity, were analyzed. RESULTS: MiR16a expression in peripheral blood of AS patients was significantly higher compared to control people (p<0.05 compared to control group). AS patients at active phase had significantly higher miR16a levels, compared to stable phase (p<0.05). Serum IL-4 and IL-10 levels in AS patients were significantly increased, while IFN-γ and TNF-α expressions were depressed (p<0.05 compared to healthy controls). MiR16a expression was positively correlated with IL-4/IL-10 or disease active index, and was negatively correlated with IFN-γ and TNF-α levels (p<0.05), but not with CRP or ESR. CONCLUSIONS: Peripheral miR16a was up-regulated in AS patients, and reflected disease activity, probably via regulating Th1/Th2 balance.

Long non-coding RNA HULC promotes proliferation and osteogenic differentiation of bone mesenchymal stem cells via down-regulation of miR-195.

OBJECTIVE: LncRNAs HULC has been reported to be important regulators in the development of various human diseases. However, the role of HULC in bone mesenchymal stem cells (BMSCs) remains unclear. The present study aimed to explore the regulatory effect of HULC on proliferation and osteogenic differentiation of BMSCs and the underlying mechanism. MATERIALS AND METHODS: The expression of HULC and miR-195 in BMSCs were altered by transfection and measured by qRT-PCR. Cell viability was measured by the CCK-8 assay. Osteogenic differentiation of BMSCs was determined by evaluation of osteogenic markers (Ocn, ALP, Runx2, and Col-1) expression levels using Western blot and qRT-PCR. Furthermore, Western blot was performed to assess the expression of proliferation-related factors, Wnt/ß-catenin and p38MAPK pathway-related factors. RESULTS: HULC overexpression significantly increased cell viability, down-regulated p21 expression but up-regulated CyclinD1 expression, and promoted the levels of osteogenic markers. However, the complete opposite effect was observed in HULC knockdown. Notably, miR-195 expression was negatively regulated by HULC and miR-195 exerted a reversed effect of HULC on BMSCs. Moreover, miR-195 mediated the regulatory effect of HULC on BMSCs proliferation and osteogenic differentiation, as miR-195 mimic abolished the effect of HULC overexpression on BMSCs. We also found that HULC overexpression enhanced the activation of Wnt/ß-catenin and p38MAPK pathway through down-regulating miR-195. CONCLUSIONS: We revealed that HULC promoted proliferation and osteogenic differentiation of BMSCs. The potential mechanism might be involved in its negative regulation on miR-195 and enhanced activation of Wnt/ß-catenin and p38MAPK pathway.

Bioinformatics analysis of common differential genes of coronary artery disease and ischemic cardiomyopathy.

OBJECTIVE: This paper aims at screening the common differential genes of coronary atherosclerotic heart disease (CAD) and ischemic cardiomyopathy (ICM), and to conduct pathway analysis and protein-protein interaction (PPI) network analysis for the differential genes. MATERIALS AND METHODS: The CAD and ICM datasets were collected from the Gene Expression Omnibus (GEO) database for human tumors to extract data components of peripheral blood RNA of patients and normal people in GSE71226 and GSE9128 chips; "limma" package of "R" software was used to screen the differential genes, and "pheatmap" package was applied to construct heat maps for the differential genes; Cytoscape, Database for Annotation, Visualization and Integration Discovery (DAVID) and String platforms were utilized for PPI network analysis, Genome Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis on the selected differential genes. RESULTS: A total of 575 differential genes were screened from GSE71226, including 350 genes with up-regulated expression and 225 with down-regulated expression, which was statistically significant (p<0.05, fold change >1). 75 differential genes were screened from GSE9128, including 47 genes with up-regulated expression and 28 with down-regulated expression. By virtue of String, DAVID and Cytoscape software, the PPI network diagram was constructed, and GO and KEGG analyses were performed successfully. CONCLUSIONS: A total of 8 common differential genes are screened, and functional annotation and pathway analysis are conducted, which is conducive to further studying the interactions between the differentially expressed genes.

MiR-299-5p targets cell cycle to promote cell proliferation and progression of osteosarcoma.

OBJECTIVE: Osteosarcoma is a common primary bone tumor with high mortality. MicroRNA (miRNA, miR) is a small RNA with 20-25 nucleotides, which could regulate diverse biological processes by targeting 3'-UTR of gene to degrade it. MiR-299-5p has been reported to participate in the progression of many diseases, but the role in osteosarcoma is still uncertain. The aim of this work was to investigate the expression of miR-299-5p in osteosarcoma and its clinical significance. MATERIALS AND METHODS: The datasets of osteosarcoma miRNA was searched in Gene Expression Omnibus (GEO) datasets, including GSE65071, GSE39040, and GSE39055. Osteosarcoma U2 and MG-63 cells were cultured in our study. Cell proliferation level after transfection was detected by using Cell Counting Kit-8 (CCK8) and colon formation assay. Cell cycles were explored using flow cytometer and cell protein expression levels after that the transfection was detected by Western blotting. RESULTS: We found that ROC curve analysis showed that miR-299-5p was a sensitivity diagnostic criteria and GSEA indicated that miRNA-299-5p may regulate cell cycle. Gain of function assay showed that miR-299-5p promotes cell cycle transition and proliferation. Reversely, the opposite results were observed with loss of function assay. At last, Western blotting assay showed that miR-299-5p may promote cell cycle transition by regulating CDK family.

Estrogen and progesterone promote breast cancer cell proliferation by inducing cyclin G1 expression.

Breast cancer is the most common cause of cancer among women in most countries (WHO). Ovarian hormone disorder is thought to be associated with breast tumorigenesis. The present study investigated the effects of estrogen and progesterone administration on cell proliferation and underlying mechanisms in breast cancer MCF-7 cells. It was found that a single administration of estradiol (E2) or progesterone increased MCF-7 cell viability in a dose-dependent manner and promoted cell cycle progression by increasing the percentage of cells in the G2/M phase. A combination of E2 and progesterone led to a stronger effect than single treatment. Moreover, cyclin G1 was up-regulated by E2 and/or progesterone in MCF-7 cells. After knockdown of cyclin G1 in MCF-7 cells using a specific shRNA, estradiol- and progesterone-mediated cell viability and clonogenic ability were significantly limited. Additionally, estradiol- and progesterone-promoted cell accumulation in the G2/M phase was reversed after knockdown of cyclin G1. These data indicated that estrogen and progesterone promoted breast cancer cell proliferation by inducing the expression of cyclin G1. Our data indicated that novel therapeutics against cyclin G1 are promising for the treatment of estrogen- and progesterone-mediated breast cancer progression.

Estrogen and progesterone promote breast cancer cell proliferation by inducing cyclin G1 expression

Breast cancer is the most common cause of cancer among women in most countries (WHO). Ovarian hormone disorder is thought to be associated with breast tumorigenesis. The present study investigated the effects of estrogen and progesterone administration on cell proliferation and underlying mechanisms in breast cancer MCF-7 cells. It was found that a single administration of estradiol (E2) or progesterone increased MCF-7 cell viability in a dose-dependent manner and promoted cell cycle progression by increasing the percentage of cells in the G2/M phase. A combination of E2 and progesterone led to a stronger effect than single treatment. Moreover, cyclin G1 was up-regulated by E2 and/or progesterone in MCF-7 cells. After knockdown of cyclin G1 in MCF-7 cells using a specific shRNA, estradiol- and progesterone-mediated cell viability and clonogenic ability were significantly limited. Additionally, estradiol- and progesterone-promoted cell accumulation in the G2/M phase was reversed after knockdown of cyclin G1. These data indicated that estrogen and progesterone promoted breast cancer cell proliferation by inducing the expression of cyclin G1. Our data indicated that novel therapeutics against cyclin G1 are promising for the treatment of estrogen- and progesterone-mediated breast cancer progression.

MiR-146a inhibits proliferation and induces apoptosis in murine osteoblastic MC3T3-E1 by regulating Bcl2.

OBJECTIVE: The dysregulation of proliferation and apoptosis plays a significant role in the pathogenesis of hormone-induced osteonecrosis of femoral head (ONFH). The research aimed to explore the regulatory role of miR-146a in dexamethasone (DEX)-induced proliferation and apoptosis change in MC3T3-E1 cells from murine osteoblastic. MATERIAL AND METHODS: In this study, MC3T3-E1 was co-cultured with 10-7 DEX for 6 h, then RT-PCR was employed to test the expression level of miR-146a and Bcl2. CCK8 assay and flow cytometry were adopted to verify miR-146a could regulate proliferation and apoptosis. After transfected MC3T3-E1 with mimics and inhibitor, RT-PCR and Western blot was used to detect Bcl2 expression level. RESULTS: In DEX treated MC3T3-E1 cells showed higher MiR-146a expression level and lower Bcl2 expression level. MiR-146a could inhibit proliferation and promotes apoptosis in murine osteoblastic MC3T3-E1 cells. Additionally, Bcl2 gene is regulated by MiR-146a. CONCLUSIONS: The MiR-146a expression level increased, while Bcl2 has low expression level in dexamethasone treated MC3T3-E1 cells. MiR-146a regulates proliferation and apoptosis of mouse bone cells. The low expression level of Bcl2 in DEX treated MC3T3-E1 cells is caused by increased MiR-146a level.

Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase

The epithelium is a highly dynamic system, which plays a crucial role in the homeostasis of the intestinal tract. However, studies on the physiological and pathophysiological functions of intestinal epithelial cells (IECs) have been hampered due to lack of normal epithelial cell models. In the present study, we established a reproducible method for primary culture of mouse IECs, which were isolated from the viable small intestinal crypts of murine fetuses (on embryonic day 19), using type I collagenase and hyaluronidase in a short span of time (≤20 min). With this method, continuously growing mouse IECs, which can be subcultured over a number of passages, were obtained. The obtained cell lines formed a tight cobblestone-like arrangement, displayed long and slender microvilli, expressed characteristic markers (cytokeratin 18 and Notch-1), and generated increasing transepithelial electrical resistance and low paracellular permeability during in vitro culture. The cells also had enzymatic activities of alkaline phosphatase and sucrase-isomaltase, and secreted various cytokines (IL-1β, IL-6, IL-8, and monocyte chemoattractant protein-1), responding to the stimulation of Escherichia coli. These results show that the primary-cultured mouse IECs obtained by the method established here had the morphological and immunological characteristics of IECs. This culture system can be a beneficial in vitro model for studies on mucosal immunology and toxicology.

A new cyclic RGD peptide dimer for integrin αvß3 imaging.

OBJECTIVE: To design a new Arg-Gly-Asp (RGD) peptide that can specifically bind integrin αvß3 and evaluate the possibility of using 131I-labeled peptide for imaging αvß3-positive tumors. MATERIALS AND METHODS: The structure of the RGD monomer was selected using V-life software. Based on the RGD monomer, a dimer of cyclic RGD [c(RGD)2] linked by Tyr-(D)Ser-Lys-(D)Ser-Ser with a Gly-Gly-(D)Ala-Gly side chain on the lysine residue was synthesized. 131I-c(RGD)2 was synthesized using the chloramine-T (ChT) method, and the octanol-water partition coefficient was experimentally measured. To evaluate its binding affinity and selectivity, its equilibrium dissociation constant (Kd) with U87 MG glioma cells was measured in vitro, while whole body imaging and biodistribution were assessed in vivo in mice bearing U87 MG xenografts. RESULTS: The optimal structure of the monomer was cyclic [-Cys-Arg-Gly-Asp-(D)Ser-Cys-]. The 131I-c(RGD)2 molecule exhibited good stability and was highly hydrophilic. The Kd value was (3.87 ± 0.05) × 10(-9) M, suggesting a high αvß3-binding affinity and specificity. The tumors were clearly visualized at 3 and 6 h post-injection. Biodistribution data of the 131I-c(RGD)2 molecule showed rapid clearance from the blood and predominant accumulation in the tumor and kidney. The tumor-to-normal tissue (T/NT) ratio increased over time. At 24 h post-injection, the tumor-to-liver, tumor-to-muscle, and tumor-to-blood ratios were 4.92, 4.29, and 5.00, respectively. CONCLUSIONS: These results suggest that the 131I-c(RGD)2 molecule may serve as a promising tracer for the detection of αvß3-positive tumors.

Glial cell line-derived neurotrophic factor promotes proliferation of neuroglioma cells by up-regulation of cyclins PCNA and Ki-67.

OBJECTIVE: We wished to test whether glial cell line-derived neurotrophic factor (GDNF) stimulates proliferation of gliomas by up-regulating expression of nuclear cyclins PCNA and Ki37. MATERIALS AND METHODS: As a model, we tested rat C6 glioma cell line exposed to basal conditions, vehicle control, or exogenous GDNF at different concentrations (0-90 µg/L) or different times (0-72 hours). Cell proliferation was quantified by MTT test, cell cycle by flow cytometry and propidium iodide staining, expression of PCNA and Ki67 by intracellular antibody staining and flow cytometry. RESULTS: We first observed that cell proliferation was most stimulated by GDNF at concentration of 70 µg/L and incubation time of 48 hours. Using this concentration and incubation time, we next documented that GDNF increased the percentage of cells in the S phase (47.98% vs. 32.57% in basal cells; p < 0.05), while not affecting the percentage of cells in G0/G1 or G2/M phases. Finally, we demonstrated that expression of both PCNA and Ki67 was significantly increased in cells exposed to GDNF. CONCLUSIONS: We demonstrate that GDNF stimulates proliferation of glioma cells by up-regulating expression of cyclins PCNA and Ki-67.

Radioactive iodine labeling of monoclonal antibody against Hsp90α and its use in diagnostic imaging in prostate cancer xenograft model.

OBJECTIVE: Heat shock protein (Hsp90) resides exclusively in the cytosol in normal cells, but is activated and then removes to the cell surface in tumor cells. The detecting upregulation or activation of Hsp90 is an early indicator of malignant behavior of cancer cells. Hsp90 has emerged as an important target for diagnosis or therapy of prostate cancer. In this study, we labeled Hsp90α specific monoclonal antibody (Hsp90α-mAb) with radioiodine Na131I and investigated its potential usage in diagnostic imaging of prostate tumor in xenograft mice model. METHODS: Hsp90α-mAb was radioiodinated by using chloramine-T. The radiolabeling efficiency and radiochemical purity were assessed in vitro. 131I-Hsp90α-mAb was then injected into the nude mice bearing human prostate carcinoma. The planar gamma Imaging was performed at 3, 6, 9 and 12 h after injection. RESULTS: The radiochemical purity of 131I-Hsp90α-mAb exceeded 95% after purification. This radiolabeled mAb was stable in human blood serum. In planar gamma imaging study, the prostate tumors in mice model were imaged clearly at 3h after injection of 131I-Hsp90α-mAb. CONCLUSIONS: The results suggest that 131I-HSP90α-mAb could be a new promising molecular probe for diagnostic imaging of prostate tumors.

Anti-tumor effect of cactus polysaccharides on lung squamous carcinoma cells (SK-MES-1).

BACKGROUND: Cactus polysaccharides are the active components of Opuntia dillenii which have been used extensively in folk medicine. In this study, we investigate the anti-tumor effect of cactus polysaccharides on lung squamous carcinoma cells SK-MES-1. MATERIALS AND METHODS: The inhibitory effect of Cactus polysaccharides on lung squamous carcinoma cells were detected by MTT assay. Cell cycle was determined by flow cytometry and cell apoptosis was determined by AnnexinV assay. Western-blotting was applied to detect P53 and PTEN protein expression in the cells treated with cactus polysaccharides. RESULTS: Results showed that different concentrations of wild cactus polysaccharides prevent SK-MES-1 cells growth and induces S phase arrest. The data also revealed that cactus polysaccharides cause apoptosis in SK-MES-1 cells determined by Annexin-V assay. Furthermore, cactus polysaccharides induced growth arrest and apoptosis may be due to the increase of P53 and phosphatase and tension homolog deleted on chromosome ten (PTEN) protein. CONCLUSION: Cactus polysaccharides have anti-tumor activity on lung squamous carcinoma cells.

Reprogramming the fate of human glioma cells to impede brain tumor development.

Malignant gliomas, the most common solid tumors in the central nervous system, are essentially incurable due to their rapid growth and very invasive nature. One potential approach to eradicating glioma cells is to force these cells to undergo terminal differentiation and, in the process, to irreversible postmitotic arrest. Here, we show that neurogenin 2 (NGN2, also known as NEUROG2) synergizes with sex-determining region Y-box 11 (SOX11) to very efficiently convert human glioma cells to terminally differentiated neuron-like cells in both cell culture and adult mouse brains. These cells exhibit neuronal morphology, marker expression, and electrophysiological properties. The conversion process is accompanied by cell cycle exit, which dramatically inhibits glioma cell proliferation and tumor development after orthotopic transplantation. Most importantly, intracranial injection of NGN2- and SOX11-expressing virus into the tumor mass also curtails glioma growth and significantly improves survival of tumor-bearing mice. Taken together, this study shows a simple and highly efficient strategy for reprogramming malignant glioma cells into postmitotic cells, which might be a promising therapeutic approach for brain tumors.

Natural products with antitumor activity from endophytic fungi.

Endophytic fungi are a seemingly inexhaustible source of novel bioactive natural products. Currently, more than 140 fungal metabolites have shown confirmed activity in tumor cell line bioassays. We present the chemical structures of these antitumor metabolites, their corresponding fungal endophytes and host plants, and the activities they exhibited, and briefly discuss some of their action mechanisms. This review emphasizes the role of endophytic fungi as an important source of leads for drug discoveries.

Cloning, expression and location of RNase9 in human epididymis.

BACKGROUND: Mammalian spermatozoa become fully motile and fertile during transit through the luminal fluid of the epididymis. At least 200 proteins are present in the epididymal lumen, but the potential roles of these luminal proteins in male fertility are unknown. Investigation of the function of these proteins will elucidate the mechanism of sperm maturation, and also provide new drug targets for male contraception. We cloned RNase9 from a human epididymis cDNA library for characterization and analysis of its functions. FINDINGS: It was predicted that human RNase9 gene was located on chromosome 14q11.2 and encoded a 205 amino acids protein with a signal peptide of 26 amino acids at the N-terminus. The protein had eight conserved cysteine residues characteristic of the RNase A family members and several potential post-translational modification sites.At the transcriptional level, RNase9 was expressed in a wide variety of tissues, and the expression was higher in men than in boys. RNase9 was localized to the post-equatorial region of the sperms' head. Immunofluorescence staining showed that RNase9 protein was present mostly in the epithelium of the epididymal tubule. Recombinant RNase9 had no ribonuclease activity. In addition, RNase9 had no detectable effect on sperm motility and fertilization as demonstrated by blocking spermatozoa with anti-RNase9 polyclonal serum. CONCLUSION: RNase9 is expressed in a wide variety of tissues. It is located on the post-equatorial region of the sperm head and the epithelium of epididymal tubule. Although RNase9 belongs to the RNase A family, it has no ribonuclease activity.

Evaluation of clinical and serological data from Taenia solium cysticercosis patients in eastern Inner Mongolia Autonomous Region, China.

Neurocysticercosis (NCC) is one of the major causes of neurological disease in China. ELISA and immunoblotting using glycoproteins purified by preparative isoelectric focusing were used to detect human cysticercosis in Tongliao area, Inner Mongolia, China in 1998. Approximately 89% (39 of 44 inpatients and outpatients with suspected NCC at Tongliao City Hospital) were residents of Inner Mongolia. About 53% were male and 77% were of working age (18-59 years), and 32% were farmers. Immunoblotting and ELISA showed a high correlation. Of the 44 patients, 31 positive by cerebral computed tomography (CT) scan were confirmed serologically to have cysticercosis. In the ELISA, patients with no lesions by CT scan had lower OD values, similar to those of normal serum. These findings confirm that both ELISA and immunoblotting assays are sufficiently sensitive to detect asymptomatic or symptomatic cysticercosis patients.