This study is to investigate the expression of CyclinD1 in asthmatic rats and construct expression plasmids of sense and antisense CyclinD1 gene and transfect them to asthmatic airway smooth muscle cell to study the effects of CyclinD1 on the proliferation of airway smooth muscle cells in asthmatic rats. CyclinD1 cDNA was obtained by RT-PCR of total RNA extracted from the airway smooth muscle in asthmatic rats. The sequence was inserted into eukaryotic expression vector pcDNA3.1 (+) to recombinate the sense and antisense pcDNA3.1-CyclinD1 eukaryotic expression vector. The two recombinations and vector were then separately transfected into airway smooth muscle cell in asthmatic rats by using liposome. The expression level of CyclinD1 was certificated by Western blotting analysis. The proliferations of ASMCs isolated from asthmatic rats were examined with cell cycle analysis, MTT colorimetric assay and proliferating cell nuclear antigen (PCNA) immunocytochemical staining. Results showed (1) Compared with control group, the content of CyclinD1 was significantly increased; (2) It was comformed by restriction endonucleasa digestion and DNA sequence analysis that the expression plasmid of sense and antisense CyclinD1 were successfully recombinated. There was significant change of CyclinD1 expression between vector and sense CyclinD1 transfected cells, and the expression level of CyclinD1 in ASMC transfected with antisense CyclinD1 was lower than that in vector transfected cells (P <0.01); (3) In the asthmatic groups, compared with the vecter group, the percentage of S + G2M phase, absorbance A value of MTT and the expression rate of PCNA protein in ASMC transfected with pcDNA3. 1-CyclinD1 vector significantly increased. The values decreased remarkably in the pcDNA3,1-as CyclinD1 group. Statistical analysis revealed that there were significant differences in these indicators of cell proliferation in three groups (P <0.01). In the normal groups, statistical analysis revealed that there were significant differences in the percentage of S + G2M phase, a value of MTT and the expression rate of PCNA protein in three groups (P <0.01). Sense CyclinD1 eukaryotic expression vectors could have a positive effect on the proliferation of ASMC, however the antisence one have a negative effect, which implicated that CyclinD1 might contribute to the process of airway smooth muscle cell proliferation.
Animals ; Asthma ; pathology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Codon ; genetics ; pharmacology ; Cyclin D1 ; agonists ; antagonists & inhibitors ; genetics ; DNA, Antisense ; genetics ; pharmacology ; Disease Models, Animal ; Gene Expression ; Genetic Vectors ; genetics ; Male ; Myocytes, Smooth Muscle ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley ; Recombination, Genetic ; genetics ; Respiratory System ; Reverse Transcriptase Polymerase Chain Reaction ; Transduction, Genetic ; Transfection

BACKGROUNDAirway smooth muscle (ASM) is suspected to be a determining factor in the structural change of asthma. However, the role of protein kinase C alpha (PKCalpha) and cyclin D1 involved in the dysfunction of ASM leading to asthmatic symptoms is not clear. In this study, the central role of PKCalpha and cyclin D1 in ASM proliferation in asthmatic rats was explored.

METHODSThirty-six pathogen-free male Brown Norway (BN) rats were randomly divided into 2 groups: control groups (group N1, N2 and N3) and asthmatic groups (group A1, A2, and A3). Groups A1, A2 and A3 were challenged with ovalbumin (OA) for 2 weeks, 4 weeks and 8 weeks respectively. Control animals were exposed to an aerosolized sterile phosphate buffered saline (PBS). The ASM mass and nucleus numbers were studied to estimate the degree of airway remodeling by the hematoxylin-eosin staining method. PKCalpha and cyclin D1 expression in the ASM cells was detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The relation between PKCalpha and cyclin D1 was assessed by linear regression analysis. PKC agonist phorbol 12-myristate 13-acetate (PMA), PKC inhibitor Ro31-8220 and an antisense oligonucleotide against cyclin D1 (ASOND) were used to treat ASM cells (ASMCs) obtained from the 2 weeks asthmatic rats. The cyclin D1 protein expression level was detected by Western blotting.

RESULTSCompared with the control group, the PKCalpha and cyclin D1 mRNA levels were increased in the asthmatic group. Similar to RT-PCR results, immunohistochemistry analysis for PKCalpha and cyclin D1 expression revealed an increased production in ASMCs after allergen treatment for 2, 4 and 8 weeks compared with the respective control groups. No difference in expression of PKCalpha and cyclin D1 in ASM were found in the 2, 4 or 8 weeks asthmatic rats. There were significant positive correlations between PKCalpha and cyclin D1 expression, both transcriptionally (r = 0.944, P < 0.01) and translationally (r = 0.826, P < 0.01), in ASM. The content of cyclin D1 in asthmatic ASMCs increased after being stimulated by PMA, and decreased when induced by Ro31-8220. ASOND targeting for cyclin D1 lowered the expression of cyclin D1 induced by PMA.

CONCLUSIONSIncreased expression of PKCalpha and cyclin D1 in ASM along with smooth muscle structure changes might implicate PKCalpha and cyclin D1 participation in the proliferation of ASM and contribute to the pathogenesis of asthma after repeated allergen exposure in rats. The results suggested that cyclin D1 might be downstream of PKC signal transduction pathway.

Animals ; Asthma ; pathology ; Cell Proliferation ; Cyclin D1 ; genetics ; physiology ; Lung ; pathology ; Male ; Myocytes, Smooth Muscle ; pathology ; Protein Kinase C-alpha ; genetics ; physiology ; RNA, Messenger ; analysis ; Rats ; Rats, Inbred BN

OBJECTIVETo explore the abnormal expression of plasma proteins by analysis of proteomic expression profile in children with infectious mononucleosis (IM).

METHODSTwo dimensional gel electrophoresis (2-DE) followed by the mass spectrometry was used to examine important protein spots with different expression levels between children with IM and normal controls.

RESULTSSeven differential proteins were obtained: hemopexin, vitamin D binding protein, fetuin A, C-reactive protein, apolipoprotein A, haptoglobin and transthyretin. Compared with the control group, haptoglobin showed a higher expression level in children with IM, and the expression levels of the other proteins were obviously down-regulated.

CONCLUSIONSThe expression changes of differential proteins identified in this study are all related with the liver acute injury, suggesting that children with IM are associated with acute liver injury. Further studies on the characteristics of above proteins will contribute to the diagnosis and treatment of pediatric IM.

Blood Proteins ; analysis ; Child ; Child, Preschool ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Infectious Mononucleosis ; blood ; Male ; Proteomics ; methods

OBJECTIVESTo investigate the prevalence of oncogenic type of human papillomavirus (HPV) infection and identify the high risk population for conducting immuno/chemoprevention of cervical cancer.

METHODSAll married women aged 30 to 50 with no history of hysterectomy, pelvic radiation and non-pregnant from certain villages of Xiangyuan and Yangcheng County were invited. This study was conducted through two phases. In phase one, subjects sampled the vaginal secretions using the collectors after signing the informed consent. And physicians sampled exfoliated cells from cervix in the phase two. All the specimens were tested with the Hybrid Capture 2 test. The data was managed and analyzed by VFP and SPSS software.

RESULTSThere were 9,683 women participated in this study. Local women welcomed this study and population compliance rate was 75.4%. In tested population, we found 2,666 subjects of HPV DNA positive and HPV prevalence was 27.5%. The rates of different age group were 24.5% (30-34 yrs), 27.4% (35-39 yrs), 28.2% (40-44 yrs), 27.4% (45-50 yrs) respectively and had no significant differences (P = 0.604). The rates were slightly increased with the higher education level and had no differences (P = 0.106). The rate in mountain areas was higher than that in half-mountain areas (P = 0.001).

CONCLUSIONSThe prevalence of HPV infection is indeed high in this region. Local women and health professionals welcome the activities of cervical cancer screening and prevention. It is an emergent task to improve their sanitary condition and prevent them from cervical cancer in these women. A women health cohort is established successfully among high HPV exposed women in rural China. The extensive biologic specimen repository has been successfully established to simultaneously study the etiology, early detection, and immuno/chemoprevention of cervical cancer.

Adult ; China ; epidemiology ; DNA, Viral ; analysis ; Female ; Humans ; Mass Screening ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; epidemiology ; prevention & control ; Prevalence ; Tumor Virus Infections ; epidemiology ; prevention & control ; Uterine Cervical Neoplasms ; epidemiology ; prevention & control ; virology ; Vaginal Smears

Objective To evaluate the efficacy and safety of endovascular repair with covered stent in treating ruptured carotid artery pseudoaneurysms.Methods The clinical data of 15 patients with ruptured carotid artery pseudoaneurysm,who were admitted to Sun Yat-Sen Memorial Hospital of Zhongshan University during the period from July 2009 to July 2015 to receive emergency rescue with endovascular covered stent implantation,were retrospectively analyzed.Of the 15 patients,the rupture of common carotid artery pseudoaneurysm caused by direct trauma was seen in one and caused by cervical operation was observed in one,the rupture of internal carotid artery pseudoaneurysm caused by cervical operation was found in 3,and the rupture of internal carotid artery pseudoaneurysm due to radiotherapy for nasopharyngeal carcinoma was seen in 10.Results Successful rescue with endovascular repair using covered stent implantation was obtained in all 15 patients.No immediate procedure-related complications or death occurred.The mean follow-up time was 11 months.During the follow-up period,no re-bleeding or cerebral ischemic complications occurred.Conclusion For the treatment of ruptured carotid artery pseudoaneurysms,endovascular repair with covered stent is minimally invasive,safe and effective with less complications;and along with the occlusion of carotid artery rupture,the blood supply of the head and neck returns to normal.However,further clinical researches with larger samples are needed before its long-term efficacy can be completely clarified.

OBJECTIVEThe effects of lentivirus-mediated suppression of Cyclin Y (CCNY) expression on the proliferation of laryngeal cancer cells were investigated in vitro.

METHODSThe lentivirus vectors containing a small hairpin RNA (shRNA) to target CCNY were constructed.Hep-2 cells were divided into the following two experimental groups:the negative control group (control lentivirus infected cells) and CCNY knockdown group (CCNY shRNA-expressing lentivirus infected cells). After Hep-2 cells were infected, Real-time PCR was used to measure CCNY expression. The influence of CCNY on the proliferation of laryngeal cancer cells were assessed using MTT and colony formation experiments.Each experiment was performed in triplicate and repeated three times.

RESULTSLentiviruses expressing shRNA against CCNY were constructed and Hep-2 cells were infected with above mentioned lentivirus at MOI (Multiplicity of infection) of 120.Real-time PCR analysis showed that the mRNA expression of CCNY in Hep-2 cells in the knockdown group was significantly decreased (P < 0.05); the mRNA level of CCNY was 75.3% lower in the si-CCNY group than in the si-CTRL group. After 5 days of lentiviral infection, the cell viability was significantly lower in cells infected with the CCNY-shRNA lentivirus compared to cells infected with the control lentivirus following a 6-day incubation. The colony number was decreased by 60% in Hep-2 cells infected with the CCNY-shRNA-lentivirus infected cells following a 10-day incubation.

CONCLUSIONSThe results suggested that lentivirus-mediated downregulation of CCNY expression decreased the proliferation and growth potency of laryngeal cancer cells.Lentiviruses delivering shRNA against CCNY may be a promising tool for laryngeal cancer therapy.

Cell Line, Tumor ; Cell Proliferation ; Cyclins ; Humans ; Laryngeal Neoplasms ; metabolism ; Lentivirus ; genetics ; RNA, Small Interfering ; genetics

Humans ; Child ; Neurofibromatosis 1/drug therapy* ; Benzimidazoles/therapeutic use*

Objective:To explore the status of disability, and characteristics of unmet needs and services of rehabilitation for children with disabilities (CWDs). Methods:A total of 130 290 CWDs administration data of unmet needs and services of rehabilitation at provincial level had been sampled and analyzed. Results:CWDs accounted for 5.33% of the total population with disabilities, in which, 60.4% were boys and 39.6% were girls; 16.3% aged 0 to nine years and 83.7% were seven to 18 years old; 88.4% with agricultural household and 11.6% with non-agricultural household. The distribution of disability severity from severe to mild were 28.3%, 39.5%, 17.7% and 14.6%. The reported unmet needs included assistive devices (40.3%), functional training (37.4%), nursing care (32.2%), medicine (24.9%) and surgery (5.7%). The received services involved in assistive devices (37.9%), functional training (33.7%), nursing care (31.0%), medicine (19.7%) and surgery (2.9%). Logistic Regression model showed that severities of disabilities had significant effects on reported unmet needs and received services (P < 0.001). Conclusion:Implement rehabilitation programs would be tailored to the unmet needs of rehabilitation as CWDs had functioning oriented unmet needs. It proposed to develop individualized and structured rehabilitation services to improve the accessibility and quality of rehabilitation for CWDs.

BACKGROUND: Shenyankangfu Tablet (SYKFT) is a Chinese patent medicine that has been used widely to decrease proteinuria and the progression of chronic kidney disease. OBJECTIVE: This trial compared the efficacy and safety of SYKFT, for the control of proteinuria in primary glomerulonephritis patients, against the standard drug, losartan potassium. DESIGN, SETTING, PARTICIPANTS AND INTERVENTION: This was a multicenter, double-blind, randomized, controlled clinical trial. Primary glomerulonephritis patients, aged 18-70 years, with blood pressure ≤ 140/90 mmHg, estimated glomerular filtration rate (eGFR) ≥ 45 mL/min per 1.73 m MAIN OUTCOME MEASURES: The primary outcome was change in the 24-hour proteinuria level, after 48 weeks of treatment. RESULTS: A total of 735 participants were enrolled. The percent decline of urine protein quantification in the SYKFT group after 48 weeks was 8.78% ± 2.56% (P = 0.006) more than that in the losartan 50 mg group, which was 0.51% ± 2.54% (P = 1.000) less than that in the losartan 100 mg group. Compared with the losartan potassium 50 mg group, the SYKFT plus losartan potassium 50 mg group had a 13.39% ± 2.49% (P < 0.001) greater reduction in urine protein level. Compared with the losartan potassium 100 mg group, the SYKFT plus losartan potassium 100 mg group had a 9.77% ± 2.52% (P = 0.001) greater reduction in urine protein. With a superiority threshold of 15%, neither was statistically significant. eGFR, serum creatinine and serum albumin from the baseline did not change statistically significant. The average change in TCM syndrome score between the patients who took SYKFT (-3.00 [-6.00, -2.00]) and who did not take SYKFT (-2.00 [-5.00, 0]) was statistically significant (P = 0.003). No obvious adverse reactions were observed in any group. CONCLUSION: SYKFT decreased the proteinuria and improved the TCM syndrome scores of primary glomerulonephritis patients, with no change in the rate of decrease in the eGFR. SYKFT plus losartan potassium therapy decreased proteinuria more than losartan potassium therapy alone. TRIAL REGISTRATION NUMBER NCT02063100 on ClinicalTrials.gov.