Objective To evaluate the inhibiting effect of selective and non-selective cyclo-oxygenase-2(COX-2) inhibitor on growth of colon cancer cell lines in vitro and its signal transduction pathway. Methods The proliferation of colon cancer cells was determined by MTT assay. COX-2, nuclear factor(NF)-?Bp65, extracellular-signal regulated kinase(ERK), phospho ERK(pERK) protein expressed in colon cancer cell lines were analyzed by Western blot. Activator protein(AP)-1 and NF-?B binding activity influenced by non-steroidal anti-inflammatory was detected by electrophoretic mobility sift assay. Results Aspirin and celecoxib could inhibit the proliferation of HT-29, SW480 and LS174-T cells and showed a dose-dependent manner. Western blot analysis showed that expression of COX-2, pERK, NF-?Bp65 protein in colon cancer cells were down-regulated by celecoxib or aspirin in some degree but not effect in total ERK expression. AP-1 and NF-?B binding activity could be stimulated by 20% fetal calf serum or tumor necrosis factor-?. Both aspirin and celecoxib could inhibit fetal calf serum-induced AP-1 activation in HT-29 and SW480 cells. Celecoxib could also inhibit tumor necrosis factor induced NF-?B binding activity, but aspirin had little effects on SW480 cells. Conclusion NSAIDs are able to potentially inhibit the growth of colon cell lines in vitro and the mechanism may relate to AP-1 and NF-?B signal transduction pathway.

Aim To observe the neuron nitric oxide synthase(nNOS) expression in the distal cerebrospinal fluid contacting neurons(CSF-CNs) of rat brain parenchyma, and investigate the role of CSF-CNs in the development of morphine dependence and withdrawal.Methods Male adult Sprague-Dawley rats, weighed 260?20 g,were experimented with A 3 ?l volume of 30% cholera toxin subunit B with horseradish peroxidase(CB-HRP) was injected into one of the rats′lateral ventricles to trace and locate the distal CSF-CNs of rat brain parenchyma 48 hours before the animals were killed. All animals were perfused and the relative tissue of rats′brain was removed.Frozen serial coronal sections (40 ?m) were cut. Then TMB-ST reaction procedure was used to stain the CB-HRP positive neurons,followed by immunohistochemistry double-labeling of the nNOS with CB-HRP positive neurons. The withdrawal symptoms were observed and scored. The numbers of the CB-HRP, and CB-HRP/nNOS positive neurons on the same segmental brain sections were counted.Results The withdrawal symptoms of the withdrawal group were significant, scores of all signs were significantly higher than those of the dependence groups and control group(P

Objective The ultrastructure of the distal CSF-contacting neurons in the dorsal raphe and the relationships with their surrounding tissues were observed and a new morphological evidence for their functional significance was offered. Methods We combined CB-HRP tracing with electron microscopic techniques and observed the ultrastructure of the distal CSF-contacting neurons in the dorsal raphe and the relationships with their surrounding tissues. Results 1.The CSF-contacting neurons have the general cytological structure of neurons; 2.CB-HRP mainly is localized in the trans face of the Golgi apparatus and lysosomes;3.There are two kinds of contrary direction synaptic relationships between the distal CSF-contacting neurons and the non-CSF-contacting neurons.Conclusions 1.There are no obvious differences between the CSF-contacting neurons and other neurons in ultrastructure;2.The Golgi apparatus and the lysosomes may exist the special function in take and transport the material from CSF;3.The CSF-contacting neurons can not only transport the material or deliver the information from the dorsal raphe to CSF,but also from CSF to the dorsal raphe.

Objective To investigate the clinical and pathological characteristics,diagnosis,differential diagnosis,treatment and prognosis of giant neurofibroma of penis in the child.Methods The clinical data including general data,imaging data,treatment methods,pathological characteristics of a case with giant neurofibroma of penis in a child were analyzed retrospectively and the relevant literature was reviewed.Results Gross appearance of the penile shaft neurofibroma was about 9 cm × 11 cm × 15 cm,with local ulceration.Computerized tomography scan revealed a giant mass in the penile shaft,about 9.0 cm × 10.0 cm × 13.4 cm.Partial excision of the penis was performed.Postoperative appearance of the residual penile shaft was about 2 cm long.The HE staining showed spindle cells with the red dye cytoplasm,spindle or elliptic nuclei and arranged in wavy partly.Positive immunostaining was presented with S-100 protein and Vimentin.The pathologic examination revealed a neurofibroma.There was no evidence of recurrence and the penis of the boy had normal sensation and erection by follow-up in 2 years.Conclusions Neurofibroma of penis in the child is extremely rare and the differential diagnosis of soft-tissue tumors of penis should be considered.The operative method should be individualized,the treatment goal is the complete resection;however,this goal must be weighed against detriment to functioning and the cosmetics of the involved organ.

BACKGROUND: It has been reported that in China, human bone marrow mesenchymal stem ceils are mostly harvested from adults. Studies on bone marrow mesenchymal stem cells in children are few.OBJECTIVE: To isolate and expand bone marrow mesenchymal stem cells from children, and to analyze the biological characteristics of bone marrow mesenchymal stem cells and their potential of differentiating into osteoblasts, adipocytes and neural like cells.DESIGN: Observational comparative study.SETTING: Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Experiments were performed at the Laboratory of Department of Orthopaedics of Wuhan Tongji Hospital from March to September 2006. Bone marrow mesenchymal stem cells were collected from one boy patient and two girl patients aged 5-8 years, who received pelvis osteotomy for dysplasia of the hip joint. The experimental procedures were approved by the Hospital Ethics Committee and family members of all children patients singed the informed consent.Dexamethasone, vitamin C, β-sodium glycerophosphate, 3-1sobutyl-1-methylxanthine, insulin, indometacin and butylated hydroxyanisole were bought from Sigma Company. Dimethyl sulphoxide was purchased from Amersco Company.METHODS: Bone marrow mesenchymal stem cells were cultured from mononuclear cells isolated over a Percoll gradient.Bone marrow mesenchymal stem cells were observed under an inverted phase contrast microscope. Bone marrow mesenchymal stem cells could differentiate into osteoblasts, adipocytes and neural like cells with osteoblast inductor (β-sodium glycerophosphate, dexamethasone, vitamin C), lipoblast inductor (dexamethasone, 3-isobutyl-1-methylxanthine,bovine insulin, indometacin) and serum-free medium inductor (dimethyl sulphoxide, butylated hydroxyanisole) respectively.Osteoblast marker (alkaline phosphatase, osteocalcin mRNA, calcium node), adipocyte marker (lipid droplet, PPAR γ-2mRNA) and neural ceil-like marker (nissl body, neuron specific enolase, neurofilament protein) were respectively determined by the immunohistochemical method, polymerase chain reaction and immunocytochemical method.MAIN OUTCOME MEASURES: ①Appearance and proliferation of bone marrow mesenchymal stem ceils from children,and ②determination results of osteoblast, adipocyte and neural cell markers.RESULTS: ①Children bone marrow mesenchymal stem cells could easily adhere to the wall, appeared fusiform, had high reproductive activity and arranged vortically after fusing. ②Appearance of bone marrow mesenchymal stem cells changed after receiving inductor. Osteoblast marker, adipocyte marker and neural cell-like marker were positive after chemical staining, polumerase chain reaction and immunocyte staining.CONCLUSION: Children bone marrow mesenchymai stem cells show stable proliferation, passage and multi-direction differentiation towards osteoblasts, adipocytes and neural like cells.

This study was conducted to evaluate the growth and NAG-1 gene expression effected by Non-steroidal anti-inflammatory drug (NSAID) on colon cancer cell lines in vitro. The proliferation of colon cancer cells were determined by MTT assay and COX-2 protein expression were detected by Western blot. Total RNA was isolated from three kinds of colon cancer cell lines; the expressions of NAG-1 mRNA in the cells treated with or without NSAIDs were assessed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay. Celecoxib, meloxicam and aspirin were able to inhibit the growth of HT-29, SW480 and LS174-T cells in dose-dependent manner. COX-2 protein expressed in HT-29 and LS174-T, but not in SW480 cells. All of colon cancer cells expressed NAG-1 gene and the level of LS174-T was lower than that of the other two cell lines. NAG-1 expression was increased by treatment with some NSAIDs in all three kinds of colon cancer cells. NSAIDs were able to potentially inhibit the growth of colon cell lines. Induction of NAG-1 gene expression by NSAID was not consistent with COX-2 expression.
Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Antineoplastic Agents ; pharmacology ; Aspirin ; pharmacology ; Celecoxib ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; metabolism ; Gene Expression Regulation, Neoplastic ; HT29 Cells ; Humans ; Neoplasm Proteins ; metabolism ; Pyrazoles ; pharmacology ; RNA, Messenger ; metabolism ; Sulfonamides ; pharmacology ; Thiazines ; pharmacology ; Thiazoles ; pharmacology

【Objective】 To explore the application value of different language assessment tools in the assessment of language development of 12-month-old high-risk infants, and to screen out simple and valid language assessment tools. 【Methods】 A total of 217 11- to 13-month-old high-risk infants who were followed up at the outpatient service for high-risk infants at the child health clinic of Guiyang Maternal and Child Health Hospital from March 2022 to May 2023 were selected as the study subjects. Their language was evaluated by Early Language Milestone Scale (ELMS), Putonghua Communicative Development Inventory (PCDI) and Ages and Stages Questionnaire-Third Edition (ASQ-3). With Gesell as the gold standard for the assessment of language, the area under receiver operating characteristic curve (AUC), sensitivity, specificity, accuracy, positive predictive value, negative predictive value, Youden index and Kappa value of the three tools were calculated. Spearman correlation analysis was used to analyze the correlation between the different language assessment scales. The Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) was used to evaluate the three tools. 【Results】 1)Among 217 high-risk infants, 78 preterm infants was the most (35.94%). The rate of delayed language development detected by Gesell, ELMS, PCDI-comprehension, PCDI-expression, ASQ-3 were 5.5%, 7.8%, 36.4%, 30.0% and 11.5%, respectively. 2)ASQ-3 had the strongest correlation with Gesell language region (rs=0.607, P<0.01) and it had the most AUC (AUC=0.812, P<0.05). The consistency between ASQ-3 and Gesell was moderate (Kappa=0.56, P<0.01). ASQ-3 had the highest sensitivity(91.7%), accuracy(93.1%), negative predictive value(99.5%) and Youden index(0.85), and ELMS had the highest specificity(94.6%). 3) Comprehensive evaluation of three tools by the TOPSIS indicated that ASQ-3 was the best, followed by ELMS and PCDI-comprehension was the worse. 【Conclusion】 Among the three assessment tools, ASQ-3 has the highest value in assessing the language development of 11- to 13-month-old high risk infants, and it may be necessary to expand the age range and establish a national norm of ELMS and PCDI in the future.

Objective: To investigate the efficacy and safety of the new investigational drug pegylated interferon α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 µg/week) combined with ribavirin in the treatment of patients with genotype 1/6 chronic hepatitis C (CHC), with standard-dose Peg-IFN-α-2a combined with ribavirin as a positive control. Methods: A multicenter, randomized, open-label, and positive-controlled phase III clinical trial was performed. Eligible patients with genotype 1/6 CHC were screened out and randomly divided into Peg-IFN-α-2b(Y shape, 40kD) group and Peg-IFN-α-2a group at a ratio of 2:1. The patients in both groups were given oral ribavirin for 48 weeks in addition and then followed up for 24 weeks after drug withdrawal. Abbott Real Time HCV Genotype II was used to determine HCV genotype, and Cobas TaqMan quantitative real-time PCR was used to measure HCV RNA level at 0, 4, 12, 24, 48, and 72 weeks. Adverse events were recorded in detail. The primary efficacy endpoint was sustained virological response (SVR), and a non-inferiority test was also performed. Results: A total of 561 patients with genotype 1/6 CHC were enrolled, among whom 529 received treatment; 90.9% of these patients had genotype 1 CHC. The data of the full analysis set showed that SVR rate was 69.80% (95% CI 65.00%-74.60%) in the trial group and 74.16% (95% CI 67.73%-80.59%) in the control group (P = 0.297 0). The data of the per protocol set (PPS) showed that SVR rate was 80.63% (95% CI 76.04%-85.23%) in the trial group and 81.33% (95% CI 75.10%-87.57%) in the control group (P = 0.849 8), and the 95% CI of rate difference conformed to the non-inferiority standard. The analysis of the PPS population showed that of all subjects, 47.9% achieved rapid virologic response, with a positive predictive value of 93.8%. The incidence rate of adverse events was 96.30% in the trial group and 94.94% in the control group, and the incidence rate of serious adverse events was 5.13% in the trail group and 5.06% in the control group. Conclusion In the regimen of Peg-IFN-α combined with ribavirin for the treatment of genotype 1/6 CHC, the new investigational drug Peg-IFN-α-2b(Y shape, 40 kD) has comparable clinical effect and safety to the control drug Peg-IFN-α-2a.