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.

Efficacy and safety of minimal pedicle screw fixation for thoracolumbar fractures: a meta-analysis.

OBJECTIVE: Minimal and open pedicle screw fixation procedures have been widely used in the treatment of thoracolumbar fractures. However, the efficacy and safety of these approaches remain unclear. This meta-analysis was conducted to evaluate perioperative, functional and radiological outcomes of percutaneous versus open pedicle screw fixation for thoracolumbar fractures. MATERIALS AND METHODS: To obtain relevant literature, a systematic search was performed using the MEDLINE, EMBASE, and Cochrane databases. The Cowley criteria were used to evaluate the risk of bias for the included studies. A database that included patient demographic information and perioperative outcomes was established. Summary odds ratios (ORs) and weighted mean differences (WMDs) with 95% confidence intervals (CIs) were estimated. Analyses were performed for the two subgroups of Chinese studies and studies from other nations. Publication bias was assessed using the funnel plot method. RESULTS: Eleven comparative observational studies that satisfied our inclusion criteria were identified via a literature search in the MEDLINE, EMBASE, and Cochrane databases. Relative to the open approach, the minimal approach was associated with less blood loss (WMD=-218.10, 95% CI: -266.31 to -169.88, p<0.00001) and shorter operative time (WMD=-15.31, 95% CI: -24.73 to -5.88, p=0.001). Evidence indicated that a significant difference was observed between Chinese studies and other studies with respect to blood loss (p=0.02). We also found that the minimal approach was associated with a lower postoperative visual analog scale (VAS) score (WMD = -1.06, 95% CI: -1.32 to -0.8, p<0.00001) and less correction loss (WMD=-0.59, 95% CI: -1.16 to 0.02, p=0.04) than the traditional open approach. No significant difference between these approaches was found with respect to complication rate (OR 0.78, 95% CI: 0.39 to 1.55, p=0.48). CONCLUSIONS: The evidence indicated that the minimal approach had better functional and radiological outcomes than the open approach. Neither approach was superior with respect to complication rate. Relative to the open approach, the minimal approach might be associated with decreased operative time, less blood loss and a shorter hospital stay.

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.

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.

Enteric glial cell activation protects enteric neurons from damage due to diabetes in part via the promotion of neurotrophic factor release.

BACKGROUND: Diabetes can result in pathological changes to enteric nervous system. Our aim was to test the dynamic changes of enteric neurons and identify the role of enteric glial cells (EGCs) in regulating enteric neuron expression in diabetic rats. METHODS: A single injection of streptozotocin (STZ) was used to establish diabetic rats. Animals were randomly distributed into diabetic 1-, 4-, 8-, and 16-week groups, as well as age-matched control groups. The PGP9.5- and glial fibrillary acidic protein (GFAP)-immunopositive cells were quantified by immunohistochemistry. The protein levels of PGP9.5, ChAT, nNOS, S-100ß, and c-fos were determined by western blotting. The levels of nerve growth factor (NGF), neurotrophin 3 (NT-3), and glial cell-derived neurotrophic factor (GDNF) were tested by ELISA. KEY RESULTS: An increase in blood glucose and a decrease in body weight were observed following STZ administration. PGP9.5 expression did not change in the diabetic ileum. However, ChAT increased after 16 weeks, and nNOS decreased after 8 and 16 weeks in the ilea of diabetic rats. The absence of degeneration of enteric neurons during the acute stage of the disease could be the consequence of the up-regulation of GFAP, S-100ß, and c-fos. Moreover, the content of NGF, NT-3, and GDNF in the ileum increased by varying degrees after 1 and/or 4 weeks of diabetes. Using 2 co-culture models of EGCs and SH-SY5Y cells in a high glucose condition, the supportive role of EGCs was further confirmed. CONCLUSIONS & INFERENCES: Enteric glial cell activation can protect enteric neurons from damage due to diabetes in the acute stage of the disease, in part via the promotion of neurotrophin release.

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.

Recent advances in management of tuberculosis in infants.

The global tuberculosis (TB) incidence estimated by WHO is found to be 8.6 million people. Moreover, the highest TB burden worldwide is found in Asian and African countries. The disease is more prevalent in infants due to their immature immune systems. Despite this, the available diagnostic tools pose a challenge due to paucibacillary nature of the disease and difficulty in obtaining specimens. The present review article discusses the important and upcoming advancements in the management of above pathological state. The article will enlighten the new vaccinations for TB in the pipeline. Moreover, new upcoming approaches involving system biology and gene expression profiling for efficient supervision of the disease will also be highlighted.

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.

Magnesium sulfate micro air pump suction for bronchiolitis treatment in infants under two years old.

OBJECTIVE: To investigate the efficiency, clinical effects and nursing methods related to the use of magnesium sulfate micro air pump suction for treating infants under two years old suffering from bronchiolitis. PATIENTS AND METHODS: From January 2014 to September 2014, ninety-six infants with capillary bronchitis were enrolled. Patients were randomly divided into two groups: experimental group (n=49) and control group (n=47). All patients went through conventional anti-inflammatory therapy. Based on this, infants in the control group were additionally treated with intravenous drip of magnesium sulfate while patients in the experimental group were treated with magnesium sulfate micro air pump suction. We recorded all changes in blood gas and clinical scores, the residence time of symptoms and signs of bronchiolitis, and hospitalization time. Results obtained on clinical effects and adverse reactions were compared and analyzed. RESULTS: The Variations of PaO2, PaCO2, SaO2 before treatment in both groups did not show any statistically significant differences (p>0.05); while after treatment analyses demonstrated that in both groups we had an increase in PaO2 and SaO2 and a decrease in PaCO2. The increase in PaO2 and SaO2 values were more pronounced while the decrease observed in PaCO2 was more significant in our experimental group. The total effective rate was significantly higher while the total adverse reaction rate, the resolution time of clinical symptoms and hospitalization time were significantly lower in our experimental group. CONCLUSIONS: Magnesium sulfate micro air pump suction was safe and effective in treating with bronchiolitis of infants below 2 years old, and its adverse reaction rate was low, nursing procedure was simple, and nursing difficulty level was low.

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.

Clinical effect of treating secondary asthma attacks of children Mycoplasma pneumoniae with combined therapy of montelukast and azithromycin.

OBJECTIVE: To discuss the clinical effects of treating secondary asthma attacks of children Mycoplasma pneumoniae with combined therapy of montelukast and azithromycin. PATIENTS AND METHODS: 96 children patients diagnosed with secondary asthma attacks of Mycoplasma pneumonia were enrolled in this study. They were randomly divided into two groups: the control group (n=49) and the observation group (n=47). Patients in the control group received combined therapy using azithromycin and bronchodilators or glucocorticoid, and patients in the observation group received a combined therapy of montelukast, azithromycin and bronchodilators or glucocorticoid. The lung function indexes, T lymphocyte subpopulation, cytokines levels, positive rate of lgG and lgM, asthma control rate and recurrence rate were compared between groups before and after treatment. RESULTS: The levels of V-T, t-PTEF/t-E, MTIF/MTEF and TEF25/PTEF in both groups increased after treatment, but we observed a more significant improvement in the observation group. The CD4+ and CD4+/CD8+ levels in both groups also increased after the intervention, while the level of CD8+ decreased. The IL-10, IL-17 and TGF-ß levels decreased more intensely in the observation group. DISCUSSION: The positive rate of lgG and lgM in both groups decreased significantly after the intervention. In the observation group, the asthma control rate was higher while the recurrence rate was lower. Although montelukast had little effect on improving the immune function, it was certainly beneficial for controlling the symptoms of asthma and improving the prognosis. CONCLUSIONS: Using combined therapy of montelukast and azithromycin for treating the secondary asthma attacks of children mycoplasma pneumonia can relieve immunological and inflammatory reactions and improve the lung function.

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.