Background Seed cells and scaffold material are the important aspects of corneal tissue engineering research.Adipose-derived stem cells(ASCs) are becoming the focus of seed cells research because of their wide source,powerful proliferation and differentiation abilities.As biodegradable polymer,polylactic-co-glycolic acid(PLGA) has successfully build multiple tissues and organs.Objective Present study was to ascertain the biological characteristics of the rabbit ASCs and their biocompatibility with PLGA scaffold in vitro and to provide groundwork for further study on the reconstruction of tissue engineered corneal stroma.Methods Adipose cells were isolated from lipoaspirate of New Zealand white rabbit using collagenase Ι.The cells were cultured and passaged.The generation 4 cells were inoculated to culture plate with 6 holes at the density of 3×104/cm2,3×104/cm2,3×106/cm2 respectively and cultivated in ossification inducing medium,lipoblast inducing medium and chondroblast inducing medium to identify the characteristics of the cells.The multilineage differentiated cells were identified by alizarin red staining,oil red O staining and immunoinfluorescene technique.The generation 4 cells were re-suspended with DiO influorescence fluid at the density of 1×107/ml and seeded on PLGA scaffold to fabricate cell-PLGA constructs.Quantitative analysis of cell proliferation on PLGA was detected by Hoechst DNA assay.The attachment and growth of adipose-derived stem cells on the scaffold were observed under the scanning electron microscope(SEM) and confocal microscopy in 1 day,3,7 days after seeding for the evaluation of biocompatibility between cells and PLGA.Results Primarily cultured cells reached 80%-90% confluence after 7-8 days with the fibroblast-like appearance.Adipose-derived stem cells of rabbits differentiated into osteoblast,adipocyte and chondroblast successfully,showing the positive stain for alizarin red staining,oil red O staining and immunoinfluorescene technique respectively.Proliferation of cells on PLGA scaffold went into plateau phase at 7 days after culture.SEM and confocal microscopy revealed the well-attached,spread cells along the scaffold and abundant excellular matrix both on the surface and interior pore of scaffold.Conclusion Cultured rabbit adipose cells have the ability of potential multilineage differentiation and good biocompatibility with PLGA scaffold,which could be used to construction of tissue engineered corneal stroma.

Animals ; Coal Tar ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Mice ; NIH 3T3 Cells ; drug effects

Objective To compare the safety and clinical efficacy between paclitaxel liposomes and paclitaxel in patients with advanced esophageal cancer .Methods A total of 90 patients with advanced esophageal cancer were enrolled into this study and were randomly divided into paclitaxel liposomes treatment group(treatment group) and paclitaxel treatment group(control group) . The patients of each group were treated with paclitaxel liposomes or paclitaxel 80 mg/m2 intravenously on day 1 and 8 ,and nedapla‐tin 75 mg/m2 intravenously on day 1 ,respectively .After two cycles of chemotherapy ,three weeks per period ,we evaluated the short term efficacy and adverse reactions according to the WHO standard .Results The short term efficacy between control group and treatment group showed no significance statistically (P>0 .05) .However ,the incidence of allergic reactions in the treatment group was significantly lower than control group (P<0 .05) .Conclusion Paclitaxel liposomal or paclitaxel combine with nedaplatin was effective equivalently in the treatment of patients with advanced esophageal cancer .But the allergic reactions of paclitaxel liposomal were lower than paclitaxel .It is worthy of clinical promotion .

Objective To investigate the clinical characteristics of infectious diseases of CNS.Methods Inpatients treated in the second hospital of Hebei Medical University from 1996 to 2005 were subjected to retrospective study.Results During 10 years,the hospitalization number of infectious diseases of CNS was 3929.The proportion of inpatients with viral meningitis(encephalitis),cerebral cysticercosis,tuberculous meningitis(TBM),purulent meningitis(PM)and cryptococcal meningitis(CM) was 63.8%,17.4%,7.7%,0.7% respectively.The hospitalization number of viral meningitis (encephalitis),TBM,PM and CM in the 2001—2005 5-year period was 53.5%,21.3%,35.3%,and 77.3% higher than that of the 1996—2000 period.Whereas the hospitalization number of cerebral cysticercosis decreased by 32.4% than that of the last 5 years.The peak months of viral meningitis (encephalitis)in a year were July,August and September.The peak months of TBM in a year were April, May and June.The case fatality of viral meningitis(encephalitis),cerebral cysticercosis,TBM,PM and CM was 8.1%,6.9%,22.4%,12.9%,33.3% respectively.Conclusions In the past l0 years,the incidence of common infectious diseases of CNS has increased in Shijiazhuang except cerebral cysticercoids. Our vigilance should be improved and some effective measures should be adopted to keep away infectious diseases of CNS.

Objective To integrate research training into clinical teaching effectively, the essential steps are understanding the students` demand and mastery, factors hindering learning and how to learn effectively.MethodsStudents in 7th grade in Peking Union Medical College (PUMC), graduate students and residents under clinical training, trained doctors for advanced studies from primary hospitals were surveyed with questionnaire by collecting educational background, the demand of learning medical research methodology, factors hindering learning, specific issues need in medical research methodology, learning model and assessment approaches.Results Totally 69 people were surveyed.92.75% of the respondents said they had studied medical research methodology and related courses before.However, 51 people (73.91%) answer they cannot read literatures with critical thinking.52 people (75.37%) indicated that they had difficulty in conducting research.58 (84.06%) people said it is very necessary to study the medical research methodology.No learning opportunities and enough time were the main hinders forlearning relevant knowledge.The top three high-demanding issues are sample size calculation and statistical knowledge, study design, and how to use the statistical software.44 (63.77%) people suggest e-learning with case based learning.Conclusions Integrate medical research methodology teaching into clinical teaching is in high-demand for students.E-learning is suggested with case based learning.

Objective: To analyze the epidemiological features of death comorbid with chronic kidney disease in Ningbo,and to provide reference for chronic kidney disease prevention and control. Methods: The information about death comorbid with chronic kidney disease in Ningbo from 2002 to 2017 was collected. The crude mortality,standardized mortality and the proportions of primary death causes were calculated. The annual percentage change（APC）was used to evaluate the trends of the mortality comorbid with chronic kidney disease. Results: From 2002 to 2017,the average annual crude mortality rate and standardized mortality rate of Ningbo residents comorbid with chronic kidney disease were 13.31/100 000 and 9.89/100 000,respectively. The crude mortality rate was 14.25/100 000 in males,which was higher than 12.37/100 000 in females（P<0.05）. The crude mortality rate was 14.88/100 000 in urban residents,which was higher than 12.32/100 000 in rural residents（P<0.05）. The crude mortality rates of Ningbo residents comorbid with chronic kidney disease from 2002 to 2017 showed an increasing trend and increased with age（P<0.05）. The primary causes of death comorbid with chronic kidney disease were urogenital diseases,endocrine nutritional and metabolic diseases,circulatory diseases,tumors and infectious diseases or parasitic diseases,accounting for 44.45%,24.32%,13.51%,6.50% and 3.97%,respectively. Conclusion The crude mortality rates of Ningbo residents comorbid with chronic kidney disease from 2002 to 2017 showed an upward trend. Men,the elderly and the urban residents were at high risk. The death comorbid with chronic kidney disease mainly attributed to urogenital disease.

Objective: To analyze the trend of the fall death rate in Ningbo,and to provide reference for prevention and control of fall death. Methods : The data of fall mortality in Ningbo from 2002 to 2018 was collected to calculate the crude mortality rate,standardized mortality rate,age specific mortality rate and the proportion of fall death types. Joint Point Regression was used to evaluate annual percentage change（APC）of fall mortality. Results : The average annual crude and standardized mortality rate of fall in Ningbo was 16.81/100 000 and 11.18/100 000,showing an upward trend from 2002 to 2018（P<0.05）. The standardized mortality rate of fall was 11.57/100 000 in men and 10.41/100 000 in women,and the difference was statistically significant（P<0.05）,both showing an upward trend from 2002 to 2018（P<0.05）. The standardized mortality rate of fall was 11.07/100 000 in urban residents and 11.25/100 000 in rural residents,and the difference was statistically significant（P<0.05）,both showing an upward trend from 2002 to 2018（P<0.05）. The mortality rate of fall increased with age（P<0.05）. The mortality rate of fall showed a small peak in children less than five years old（1.81/100 000）,and began to rise rapidly in people aged 65 years or over and peaked again in people aged 85 years or over. There were 11 141 cases died of falling on the same plane,accounting for 68.25%. Fall deaths occurred mainly at home,with 11 829 cases（72.46%）. Conclusion The mortality rate of fall in Ningbo increased from 2002 to 2018. The males,urban residents,people aged less than five years and aged 65 years or older have high risk of death from fall,who were targets for prevention and control strategies of fall.

Objective] To investigate influence factors for willingness to influenza and pneumonia vaccination in COPD patients, thus providing a reference for relevant health education and vaccination pro-motion. [ Methods] A total of 629 COPD patients aged 60-74 were selected as subjects from 3 dis-tricts in Ningbo City of Zhejiang Province.Questionnaire involving personal information, occupation, in-come, behavior and willingness to vaccination were used for the survey through October to November in 2013 .Multivariate unconditional logistic regression was used to analyze the influence factors in vaccination willingness. [ Results] Influenza and pneumonia vaccination rate was found to be 10.49%, and there were 36.88% of the subjects who voluntarily vaccinated against influenza and pneumonia in community hospital.Factors as high income, vaccination within 5 years contributed to higher vaccination willingness;and older age, misgivings about adverse reaction of vaccination were factors affecting the willingness to vaccination. [ Conclusion ] Extensive health education for adult vaccination should be further developed.For the older, COPD patients and other special population, influenza and pneumonia vaccination should be offered free of charge by government, so as to raise the vaccination rate.

OBJECTIVETo explore the suppression effect of human cytomegalovirus (HCMV) on megakaryocytes and their precursors and study the antiviral effect of antisense phosphorothioate deoxyoligonucleotide (ASON) against HCMV.

METHODSCD34(+) cells were induced to proliferate and differentiate committedly to megakaryocytes in a semi-solid CFU-MK culture system. Cultured cells and ASON pretreated CD34(+) cells were infected by HCMV of AD169 strain. HCMV immediate early protein (IEP) DNA and mRNA and UL36 mRNA were detected by PCR and reverse transcription-polymerase chain reaction (RT-PCR). Cytotoxicity was evaluated by MTT assay.

RESULTSHCMV AD169 suppressed the proliferation of megakaryocytes significantly. Compared with the mock group, the CFU-MK yields were decreased by 21.6%, 33.8%, and 46.3%, respectively, in 3 different titers of virus infected groups (P < 0.05). The suppression was virus titer dependent. HCMV IEP DNA, HCMV IEP mRNA and UL36 mRNA were detected in the colony cells of viral infection group. Compared with the infected group by HCMV AD169, UL36Anti treatment at 0.08 micromol/L could recover the CFU-MK yields significantly (P < 0.05). In the infected MK, which was pretreated with UL36Anti at 0.08 micromol/L, HCMV UL36 mRNA was undetectable by RT-PCR. The oligonucleotide MM(1) containing a G-to-C substitution in UL36Anti was inactive at 0.08 micromol/L but active at 0.40 micromol/L. The concentration of UL36Anti necessary to significantly affect cell growth was 90.00 micromol/L.

CONCLUSIONSHCMV AD169 infection inhibits the proliferation and differentiation of megakaryocytes and their precursors. There are early transcriptions of HCMV IE and UL36 protein in infected CFU-MK. The specific ASON has a definite anti-HCMV activity.

Antigens, Viral ; genetics ; Antiviral Agents ; pharmacology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Cytomegalovirus ; genetics ; physiology ; Fetal Blood ; cytology ; Host-Pathogen Interactions ; Humans ; Immediate-Early Proteins ; genetics ; Infant, Newborn ; Megakaryocyte Progenitor Cells ; cytology ; drug effects ; virology ; Oligonucleotides, Antisense ; genetics ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the effects of nickel-refining dusts on DNA damage in mouse NIH/3T3 cell.

METHODDNA damage of mouse NIH/3T3 cell induced by two nickel-refining dust samples was determined with single cell gel electrophoresis (SCGE) technique.

RESULTS(1) Under the condition of the same treatment time, the tailed cell (%) of NIH/3T3 cells increased with the increase in doses of nickel-refining dusts (35.5%, 69.7%, 85.2%, 41.3%, 75.7%, 89.2% respectively except for sample 2, 50 microg/ml, 24 h group), and DNA strand breaks reached the peak value at 4 h of exposure; (2) When we treated the NIH/3T3 cells with the same dose of nickel-refining dusts, the tail rate at 4 h was higher than those at 2 h and 24 h of exposure; (3) Both sample 1 and sample 2 with different doses of nickel-refining dusts could induce higher comet tail, DNA%, tail length (except for 12.5 microg/ml), extent of TM (except for sample 1, 12.5 microg/ml) than in control group (P < 0.05). The DNA damage range was significantly increased in different tested doses of nickel-refining dusts and the damage range reached the peak value when the cells were treated for 4 h.

CONCLUSIONNickel-refining dusts could induce different degree DNA damage in NIH/3T3 cells.

Animals ; Cell Cycle ; Cell Survival ; Comet Assay ; DNA Damage ; Dust ; Metallurgy ; Mice ; NIH 3T3 Cells ; Nickel ; toxicity