Objective To construct human eukaryotic expression system expressing T cell surface molecule CD40L and identify the expression product in SCCL cell line h446. Methods The expression vector pcDNA3.1-CD40L was constructed by recombinant DNA technology, then transfected into h446 cell line of SCCL by lipidosome methods, and the expressed membrane protein was identified by flow cytometry. Results The pcDNA3.1(+)-CD40L was constructed successfully and the coding region was inserted into the vector correctly. The membrane protein was observed by flow cytometry on transfected h446 cell line of SCCL. Conclusion Human CD40L was expressed successfully in eukaryotic system, which is very important in the anti-tumor vaccination study with a recombinant human CD40L in clinic.

BackgroundMüller cells has been recognized as being vital in both healthy and diseased retina.Recently,these cells even have been identified to be the source of retinal progenitor cells.In order to study the possible function of the retinal Müller cells,it is important to establish a practical procedure to obtain the purified cells. ObjectiveThis study was to simplify the procedure of primary culture and purification of retinal Müller cells in vitro. MethodsEyeballs of SPF newborn SD rats were enucleated and retinas were dissected free after soaking the eyeballs overnight in DMEM/F12 medium in room temperature.Then the retinas were mechanically dissociated into micro aggregates and cultured in DMEM/F12 medium containing 10％ FBS for 8-10 days.The floating retinal aggregates and debris were removed and the medium was changed in 2-3 days interval to get more purified flat cell population.Cultured cells were passaged after confluent.Immunofluorescence staining was used to detect the response to glutamine synthetase (GS) and Vimentin for the identification of the cultured Müller cells,and flow cytometry (FCM) was used to estimate the purity of the cells. Results Cultured Müller cells had large cellular body and richer cytoplasm.More than 95％ of the cells showed the positive response for GS with the brown staining in cytoplasm and cellular nuclei,and the positive stainiug also was seen for Vimentin in cytoplasm.FCM showed that 99.7％ of the cells were GS positive after 3 passages.Conclusions Modified procedure in this experiment is a simple and practical method for culturing retinal Müller cells.

Retina is subjected to many acquired and inherited neuronal degenerative diseases such as agerelated macular degeneration (AMD), diabetic retinopathy (DR) and retinitis pigmentosa (RP). All of these diseases are associated with the progressive damage and loss of photoreceptors, which is causing visual impairment and irreversible blindness. Stem-cell therapy is being widely considered as a promising treatment of these incurable retina diseases. However, in mammals including humans, there seems to be little or no recovery of lost cells. By contrast, nonmammalian vertebrates, such as amphibians and fish, have robust regenerative responses to injury, which can lead to the near complete restoration of the neurons lost through injury. Nevertheless ,over the past several years, studies have investigated that stem cells do exist in the adult mammalian eyes, and at least some types of neurons can be regenerated in the mammalian retina by stimulating with growth factors or transcription factors. These recent results suggest that some part of the regenerative program occurring in lower vertebrates remains in the mammalian retina.Here, the origin of various of adult retinal stem cells for the self-renewal and proliferation. and the relevant influencing factors were summarized.

Objective:To analyze the status quo and problems of the health human resources of township health center.Methods:296 township health centers in 18 counties of 6 provinces were selected with typical sampling and workers in the centers as target population.Relative administrative officials and directors of township health centers were interviewed.Descriptive analysis and thematic analysis were conducted with quantitative data and qualitative da-ta.Results:The volume and distribution of human resources in township health centers are not balanced and the allo-cation standard of the middle area is low.The null establishment and the employees out of establishment coexist.The allocation of health human resources inside township health centers is not reasonable and the ratio of health profes-sionals is lower than national standard.The rate of senior professionals is low.That health workers are out of stock is a potential problem in some counties.Medical education is not matched with the demand of township health centers.The recruitment policy is inappropriate and the remuneration of health professionals is low.It is difficult for the worker in township health centers to get professional promotion.Recommendations:The structure of workers in township health centers and the recruitment policy should be adjusted.The path of professional promotion should be clear in order to ex-pand the space of career development of workers in township health centers.Improving the remuneration of workers in township health centers should be considered to increase the attractiveness of the post in township health centers.

Objective:To Compare the operational status of the New Rural Cooperative Medical Scheme( NCMS) in 14 Counties of 6 provinces. Methods:Two provinces were selected from eastern, central and western areas respec-tively and then two counties were chosen randomly from each province, but each of Jiangsu and Guangxi Provinces pro-vided 3 to reach the sample number of 14 counties. Excel 2007 was used for descriptive and comparative analysis of fund and in-patient service for NCMS. Results:The average funding standard was about 300 and 350 Yuan for 2012 and 2013. Hospitals outside county were frequently used and the ratio was over 40% in county I. Except Jiangsu and An-hui, the rate of enrollees who get compensation for inpatient service was over 10% and the actual compensation rate more than 50%, Fujian being an exception. Average hospitalization costs per time were different among counties and significantly rose in 2013. The ratio of out of pocket inpatient service expenses to the net rural household income was diverse among counties and it declines in some of them in 2013. The rate of fund for hospitals outside county was high and that of G counties was more than50%. The fund surplus rate was negative in that same year and was accumulatively ranging between 1 and 2%. Conclusions:The NCMS financing level was low and the personal financing responsibility was lighter;the enrollees didn’t contribute enough. In-patient service utilization structure was not rational in different level hospitals. The actual compensation rate for inpatient service didn’t increase a lot and the medical expenses burden didn’t alleviate apparently. The funds supervision was weak and it probably leaded to a high risk fund deficit.

Objective: To analyze the current status of village health worker practice and their remuneration in six provinces. Methods: After 18 counties (cities/districts) from 6 provinces are selected based on their levels of economic and social development, this paper uses self-designed questionnaires to survey all village clinics under the jurisdiction of these 18 counties. Results: The average numbers of village health workers range from 6. 79 to 19. 05 per 10,000 rural populations and 1. 36 to 3. 24 per village. In some provinces, more than 20% of the village health workers are 60 years old, and the educational level of more than 50% is technical secondary school and they have village health worker prac-ticing certificates only. The coverage ratio of medical malpractice liability insurance among village health workers in two western provinces is less than 11%. Except Jiangsu province, village health workers in the other five provinces have mainly joined the New Rural Pension Scheme ( NRPS) . Jiangsu province ranks the highest in subsidies for the services of public health and essential medicines per village health workers, and Fujian province ranked the lowest. Village health workers who are included in the administrative staffing management of township health centers account for less than 20%. Except Jiangsu province, more than 70% village health workers provide 24-hour service. The number of the outflow personnel is higher than the number of the inflow. Conclusions and Suggestions:The problem of village health worker aging is getting worse. Their education level, practicing quality and remuneration are low. This paper suggests the government not only improve the practicing quality and define the legal status of village health workers, but also in-crease the remuneration, incentives and job satisfaction of village health workers Further studies should be conducted in order to put forward a more practical advice about how to attract more health workers in rural areas.

Objective To observe the chronological and spatial expressions of transforming growth factor-? receptor Ⅱ(TGF-?RⅡ) and fibronectin(Fn) in uriniferous tubules of mouse kidney and investigate the correlation of TGF-?RⅡ and Fn with development of uriniferous tubules.Methods The expressions and content changes of TGF-?RⅡ and Fn were examined by immunohistochemistry and stereological method in mouse kidneys on embryonic days 12,14,16 and 18 and postnatal days 1,3,7,14,21,28 and 42.Results The expression of TGF-?RⅡ was found in all stages of renal tubules and collecting ducts and increased gradually with the kidney development;however,the expressions were obvious in all stages of proximal tubules and collecting ducts,while little was observed in all stages of distal tubules.The expression of Fn was detected in all stages of renal tubules and collecting ducts and also increased gradually with the kidney development.Conclusion TGF-?RⅡ and Fn might play an important role in the development and maturation of uriniferous tubules of mouse kidney.

In order to understand the research status of HUANG Fu-mi's A-B Classic of Acupuncture and Moxibustion, 83 pieces of literature that have been publically published since 1978 were analyzed. A summary was performed according to 7 aspects including version origin, emendation, arrangement and summary, theory discussion, clinical research, value evaluation, the rest questions, etc. It is believed that deep research work has been done on explanations, emendation and translation regarding A-B Classic of Acupuncture and Moxibustion. However, the exploration work on the theory and application is far from enough. Therefore, beginning from historical and cultural background, we should analyze the humanistic spirit of HUANG Fu-mi combined with A-B Classic of Acupuncture and Moxibustion to perform a deep exploration, so we could have a thorough and comprehensive research on HUANG Fu-mi's academic thoughts.
Acupuncture ; education ; Acupuncture Therapy ; history ; methods ; Books ; history ; China ; History, Ancient ; Humans ; Medicine in Literature ; Moxibustion ; history ; methods

Objective To observe the chronological and spatial expression of fibronectin(Fn)and TGF-?RⅡ in developing mouse glomeruli.Methods The expressions of Fn and TGF-?RⅡ were examined by immunohistochemical and stereological method in embryos(including fetal age of 12,14,16 and 18d)and postnatal mouse kidneys(including the age of 1,3,7,14,21,28 and 42d).Results Immunohistochemical results showed that at fetal age of 14d,Fn was stained in S-shaped corpuscles,but not stained in the comma-shaped corpuscles.After fetal age 16d,Fn was expressed in all stages of glomeruli except the comma-shaped corpuscles.TGF-?RⅡ was stained in all stages of glomeruli from fetal age 14d.Fn and TGF-?RⅡ were localized respectively in basement membrane and cell plasma of glomeruli.Stereological analysis showed that the expression of Fn was increased gradually in all stages of glomeruli with renal development,except in the comma-shaped corpuscles;also,the expression of TGF-?RⅡ increased gradually in all stages of glomeruli with renal development.Conclusion The expressions of Fn and TGF-?RⅡ in embryonic and postnatal mouse kidney show chronological and spatial sequence,implying that they might play a role in the development and maturation of renal glomeruli of mouse.

OBJECTIVETo modify biomacromolecules, such as chitosan and collagen, to synthesize a mineralized template that will induce self-growing remineralization of tooth enamel.

METHODSNatural polycation polysaccharide chitosan was modified through phosphorylation to synthesize the polyanion derivative ofphosphorylated chitosan. Parent hydrogels com- bined with chitosan and collagen I were built through peptide binding reaction using genipin as a crosslinker. The gels self- assembled on the tooth's inert surface, which was stimulated by ultraviolet radiation. The bionic saliva provided mineralized ion, and then the hydroxyapatite assembled and grew in situ on the tooth.

RESULTSThe functional group P04(3-) (3,446 cm(-1)) was grafted on chitosan as confirmed by the Fourier transform infrared spectroscopy. The porous polyelectrolyte complex hydrogel formed by the interaction between the polycation chitosan and the polyanion phosphorylated chitosan could induce hydroxyapatite crystal nucleation and growth on the hydrogel fiber surfaces. The neonatal crystal was hydroxyapatite as confirmed by X-ray diffraction and was tightly connected to the tooth. A continuous structure of column crystals with sizes ranging from 30 nm to 60 nm was observed. The structure was in parallel direction similar to the direction of the enamel rod, and its hardness was close to dentin.

CONCLUSIONThe parent hydrogels that were easily obtained and controlled could mimic the template of the enamel mineralization and induce a self-growing hydroxyapatite, which is an important step in the structural bionics of enamel.

Biocompatible Materials ; Chitosan ; Collagen ; Dental Enamel ; Durapatite ; Microscopy, Electron, Scanning ; Polymers ; Spectroscopy, Fourier Transform Infrared ; Tooth ; Tooth Remineralization ; Ultraviolet Rays ; X-Ray Diffraction