Retina is composed of a heterogeneous population of cell types, each with a unique biological function. Even if the same type of cells, due to genetic heterogeneity will lead to cell function differences. In the past, traditional molecular biological methods cannot resolve variations in their functional roles that arise from these differences, and some cells are difficult to define due to the lack of specific molecular markers or the scarcity of numbers, which hindered the understanding and research of these cells. With the development of biotechnology, single-cell RNA sequencing can analyze and resolve differences in single-cell transcriptome expression profiles, characterize intracellular population heterogeneity, identify new and rare cell subtypes, and more definitely define the characteristics of each cell type. It clarifies the origin, function, and variations in cell phenotypes. Other attributes include pinpointing both disease-related characteristics of cell subtypes and specific differential gene expression patterns, to deepen our understanding of the causes and progression of diseases, as well as to aid clinical diagnosis and targeted therapy.
Objective To observe the dynamic expression of nestin and glial fibrilary acidic protein (GFAP) in the development of retina in rats.Methods In 48 Wistar rats, 24 were divided into 8 groups with 3 rats in each according to their age (1 day, 1 week, and 2, 3, 4, 7, 12, and 20 weeks old). The sagittal freezing sections of the eye were made; nestin/glutamine synthetase (GS) and GFAP/GS were stained by immunofluorescence and were observed under the confocal microscope. Total RNA was extracted from 18 rats which were divided into 6 groups according to the age (1 day, 1 week, and 2, 3, 4, and 12 weeks old) with 3 rats in each. The expression of nestin, GAFA and GS mRNA were detected by realtime quantitative reverse transcription polymerase chain reaction (RT-PCR). Müller cells were cultured from postnatal day 7-12 rats; the expression of nestin and GFAP was detected by immunostaining study. Double immunofluorescence was carried out between nestin/GS and GFAP/GS.Results One day after the birth, nestin positive cells were found in the whole retinal neuroblast layers with elongated retinal progenitor cells; the GFAP positive astrocytes were observed in the inner retina. One week after the birth, Müller glial cells expressed GS and nestin but not GFAP; GFAP positive cells localized in the inner retina.Two to 12 weeks after the birth, the expression of nestin in Müller cells decreased and even disappeared; the expression of GFAP in astrocytes didn't change much. The Müller cells expressed nestin but no GFAP in vitro. The expression of nestin and GFAP mRNA in retina was accordant with the results of immunofluorescence staining.Conclusion In the developing retina, the expression of nestin in Müller cells decreases gradually, and no expression of nestin can be found in adult rats; the expression of GFAP can't be observed in Müller cells in neonatal and adult rats.
ObjectiveTo analyze the electro-clinical characteristics and surgical outcome of low-grade developmental tumors in temporal lobe. MethodsThe onset age, seizure duration, seizure types, electroencephalogram and surgical outcome of 49 patients with low-grade developmental tumor of temporal lobe were analyzed retrospectively. ResultsTwo groups of the seizure types were divided. The first group was spasm, the other was focal onset. There were 12 cases in spasm group, with an average onset age of (1.00±0.59) years. The discharge was extensive and multi-brain-area locaded, especially in the temporal montages and the ipsilateral posterior montages. There were 37 cases in second group, with an average onset age of (8.90±8.84) years, mainly including autonomic seizure, tonic seizure and automotor seizure. In this group, the discharge was mainly recorded in the temporal montages, which could spread to the frontal montages and less locaded in posterior montages. The difference of onset age between the two groups was statistically significant (P<0.01). The average follow-up of spasm group was (2.80±1.57) years, and the surgical outcome of all patients in this group were all Engel I (100.00%, 12/12). The focal onset group was followed up for an average of (6.50±4.78) years, and the rate of Engel I was 91.80% (34/37). There was no significant difference between the two groups (P>0.05). ConclusionsFor low-grade developmental tumors in temporal lobe, there are two seizure types, including spasm and focal onset. The onset age of spasm is earlier, while patients with focal onset mostly start at childhood or older, rare in infancy. Surgery has a good effect on the treatment of temporal lobe developmental tumor epilepsy.
Based-on retrieval and reviewing information of evidence-based clinical practice guidelines from the literature, this paper introduced the key components of assessment guideline quality, including the process of development and reporting of guideline.
ObjectiveTo develop the questionnaire and test its reliability for investigating route, prevention, and control of SARS-CoV-2 infection in medical staffs.MethodsThis questionnaire was development based on the COVID-19 relevant guidelines, official documents issued by the National Health Committee of the People's Republic of China, and published studies. The development group performed repeated discussions and drafted the first questionnaire, then performed expert consultation and revised the draft according to their suggestions. Eventually, some frontline medical staffs were invited to carry out pre-test investigation of the questionnaire and test its reliability.ResultsThe first draft included 48 items; 18 experts were invited in the first round questionnaire and 10 experts in the second round questionnaire. The positive coefficient of experts in these two rounds was both greater than 75%, and the authority coefficient of experts' opinions was greater than 0.70. The variation coefficient of these items was between 0.00 and 0.35, the coordination coefficient of experts was 0.193 (P<0.05). The experts of above two rounds put forward 14 suggestions for text modification or adjustment options of some items; after the development group held repeatedly discussions, a total of 8 items were performed secondary consultation and finally reached consensus. The final questionnaire included two domains of questionnaire before and after confirmed diagnosis. The domain "before confirmed diagnosis" covered 4 sections and 29 items involving infectious cause, plan and knowledge of prevention and control, and psychological symptoms. The domain "after confirmed diagnosis" covered 5 sections and 21 items, included symptoms, treatment, and psychological status after diagnosis; impact on the surrounding environment and people, and awareness of protection after infection. The pre-test results showed that the total items were considerably numerous, some items were difficult to understand, some laboratory results and treatment conditions were ambiguous, etc. After modification and re-testing, the test-re-test reliability of each domain was between 0.74 and 0.93, and the overall re-test reliability of the questionnaire content was 0.82.ConclusionsThis research has developed a questionnaire for investigating infection process, prevention and control of SARS-CoV-2 infection in medical staff, and the items considered two domains prior to and after confirmed diagnosis. The reliability and practicability of the questionnaire are acceptable.
ObjectiveTo compare the similarities and differences in procedures between Western medicine (WM) and traditional Chinese medicine (TCM) guidelines. MethodsWe systematically searched China National Knowledge Infrastructure, WanFang Data, China Biology Medicine disc and PubMed from inception to May 17, 2022. Supplementary search of reputable medical institutions and organizations, as well as retrospective references were conducted and screened. We extracted and analyzed the basic information and guideline development procedure of the included handbooks. ResultsA total of 10 handbooks published from 2013 to 2022 were included, with four WM handbooks and six TCM handbooks. The median completeness of handbooks was 53.3%, with a maximum of 93.3% and a minimum of 43.3%. There are six handbooks with less than 60.0% completeness. The median reporting rate of the key steps was 65.0%, and the reporting rate of 5/8 of the key steps was more than 80.0%. Among the key reporting steps, role of funders, update methods, and conflict of interest management were reported at a low rate. Compared with WM handbooks, TCM handbooks reflected TCM characteristics in the procedure of topic selection, working groups, conflict of interest declaration and management, clinical questions, evidence, recommendations, report guideline, and external review. ConclusionThe completeness and reporting rate of the key steps need to be improved. TCM characteristics need to be further integrated when developing TCM guideline handbooks. We highly recommend that guideline stakeholders actively participate in handbook development to promote the quality of handbooks.
High-quality development has become the command stick for the current reform of medical institutions. This article combines the practical experience of West China Hospital of Sichuan University to sort out the difficulties in the construction of provincial medical quality control centers in China. It summarizes the problems in policy support, intervention methods, work content, network construction, discipline construction, and quality control efficiency of provincial medical quality control centers, and proposes that building a collaborative platform based on key management projects, establishing a multi-level operation mode, and constructing a benchmark construction model are the key paths to reform the management of provincial medical quality control centers.
Objectives To investigate the participation of magazines or journals' editors in the clinical practice guidelines in China. Methods WanFang Data, VIP, CNKI, CBM databases, as well as Baidu, Google and www.medlive.cn were searched online to collect incorporated guidelines in which magazines or journal editors participated in. Data was then analyzed. Results In total, 68 guidelines were selected, with 51 (75.00%) led by magazines and 17 (25.00%) edited by editors. 55 guidelines (80.88%) were the same in published and participated journals. Circulatory diseases (27.94%), diagnosis and treatment (54.41%) were the most concerned. 15 guidelines (22.06%) were updated. 17 guidelines reported the roles of the editors who were mostly expert group members (13.24%). 7 guidelines, 3 of which affirmed no relevant conflicts of interest, reported the sponsorship. The quality was higher than the domestic average while lower than the international guidelines. Conclusions The number of clinical practice guidelines magazines or editors participating in China is relatively small, while the quality was higher. The primary form of the participation is journal-led, however, the process, methods, roles, and conflicts of interest in the guidelines require further definition.
Objective To investigate effects of neural retina on development of the structure of outer blood retinal barrier in embryogenesis. Methods The retinal neural epithelium (RNE) and pigment epithelium (RPE) layers of 150, 120 and 90 embryonic chicken eyes incubated for 7,10, and 14 days were peeled off. RNE was used to prepare the culture medium with different conditions (7drcSF3, 10drcSF3, 14drcSF3). RPE cells of 7- and 14-incubated chicken embryos were cultured on laminin-coated transwell filter. The SF3, 7drcSF3, 10drcSF3 , 14drcSF3 medium were used respectively in the apical chamber and SF2 was used in basolateral chamber. After the formation of monolayer, the transepithelial electrical resistance of the RPE was detected. After the fixation of RPE cells, the condition of the tight junction among the cells was observed by immunohis tochemistry and transmission electron microscopy. Results For the RPE cells of 7-and 14-day incubated embryonic eyes, the difference of TER in various medium of SF3/SF2, 7drcSF3/SF2, 10drcSF3/SF2, 14drcSF3/SF2 was statistically significant (P＜0.01). The polarity of RPE cells was induced and the netlike tight junctional strands was urged in the retina-conditioned medium. Conclusion The neural retina may actively promote the formation of the structure of outer blood retinal barrier. (Chin J Ocul Fundus Dis，2004，20：237-240)