Objective The diolistic assay has been modified to make it simpler and more efficient in labeling neurons and glia. Methods CNS neurons and glial cells were labeled with DiI diolistic assay in fixed tissue and living brain slices of C57/B6J mice. Results The method allowed the visualization of the fine structure of neurons and glia including synaptic structures such as dendritic spines. Conclusion With the method, the labeling efficacy of cell's fine structure is improved, making it preferable for the analysis of dendritic spine. In addition, the ability to label the living neuron and glia will extend its application vastly.

Objective To understand the work tasks of rural doctors in our country and to provide bases for the reforms of clini‐cal medical education of junior colleges .Methods A self‐made questionnaire was performed in 1 245 rural doctors from 86 rural grassroots health institutions in 7 provinces/municipalies of our country .Results The academic levels of rural doctors were low (65 .54% )and the rates of their medical certification holding were low in western district(7 .79% ) .The work load including basic medical care and basic public health services they committed were approximately matching .The top 5 items of basic medical care were ,in proper sequence ,diagnoses and treatments of common diseases ,common symptoms ,chronic disease management ,communi‐ty rehabilitation ,community first aid .The top 5 items of basic public health services were establishment of residents′health files , maternal and child health ,health care of the aged ,immunization ,health education .Conclusion The technological levels of diagnosis and treatment and theoretical knowledge of rural doctors should be further improved .The clinical medical education of junior medi‐cal colleges must strengthen the training and cultivation of the students′abilities of basic public health services ,besides consolida‐ting the training of the abilities of practical diagnosis and treatment .

Objective To learn the professional ability of rural doctors in our country ,to provide scientific reference for educa-tional reform of junior-college clinical medicine .Methods A self-made questionnaire was performed in 1 245 rural doctors from 86 rural grassroots health institutions in 7 provinces(municipalities) of our country .Results The rural doctors were familiar with working environment ,coordinated and communicated well with colleagues ,basic competence at work .But their clinical diagnosis-treatment technology and operational capability of medical equipments were not strong .The doctor-patient relationship was not so good .Conclusion Not the proper use of medical equipment and treatment technology of weak seriously affected the quality of serv-ice .Not ideal doctor-patient relationship influenced the service effect .It was suggested that college of clinical medical professional education should enhance the cultivation of students′clinical diagnosis technology ,basic operational skills and communication skills between doctors and patients .

Objective To explore the effect and mechanism of Gli2 on EMT and invasion in the hepatocellular car-cinoma cell line SMMC-7721.Methods shRNA of Gli2 and Negative control (NC) shRNA were constructed and transfected into SMMC-7721 cells.shRNA-Gli2 group,shRNA-NC group and blank group were set up .Transwell chambers assay , adhesion experiments were used to detect the ability of invasion ,homogeneous and heterogeneous cells intercellular adhesion of each group .Meanwhile, the qRT-PCR, Western blot were used to examine Gli2, E-cadherin ,N-cadherin and vimentin mRNA and protein expression .Results In different hepatocellular carcinoma cell lines and be along with the increasing ability of the invasion in hepatocellular carcinoma cell lines , Gli2 expres-sion was higher .Compared with the shRNA-NC group and blank control group ,the interfered group extensive cells invasion ability was inhibited ( P <0.05 ) , homogeneous cells intercellular adhesion increased and heterogeneous cells intercellular was opposite ( P <0.05 ) , meanwhile , the expression of E-cadherin was declined significantly ( P<0.05 ) ,the expression of N-cadherin , vimentin raised significantly ( P<0.05 ) .Conclusions Silencing Gli2 gene can reverse the EMT of the hepatocellular carcinoma cell line SMMC-7721 and inhibit cell invasion ,its mecha-
nism may be related to the up-regulation of E-cadherin and the dow N-regulation of N-cadherin, vimentin.

Apoptosis ; Cell Proliferation ; Gene Expression ; Gene Silencing ; HL-60 Cells ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; RNA, Small Interfering ; genetics

The problems about scientific research design,data disposal and paper writing of medical scientific research are described in the article.

Objective To Investigation the quality of marrow smear purchased by CDUTCM.Methods The quality and the typ-icality of marrow smears purchased during 2015 -2016 were collectively examined,and then decided whether these smears fit the blood cell morphology experimental teaching requirement.Results Of all the 960 marrow smears purchased these two years, 49.7% failed in smear made or stained,and 16.0% failed to meet the teaching requirements in the typicality of marrow cells.Con-clusion Teaching marrow smears,being different from clinic ones in their preparation and morphological diagnosis,must be of great quality in sustaining and of better typicality in their cell features.

Objective:To detect the expression of tumor necrosis factor alpha in (TNF alpha) breast cacner and its relationship with imaging features.Methods:Using immunofluorescence and flow cytometry to detect the expression of TNF-α in MDA-MB-231 and MCF-7.Collect 82 patients with mammary gland disease,which was confirmed by pathological tissue,its pathological data,imaging data,and by immunohistochemistry to detect the expression of TNF-α in breast tissues,and analyze and the relationship between its expression and the pathological features and imaging characteristics.Results:TNF-α high expression in MDA-MB-231,the expression of TNF-oα in malignant breast tumor tissue significantly higher than that in benign tumor,the expression quantity associated with lymph node metastasis,TNM stages,strengthen uniform in MRI,the boundary and the shape of the X-ray Mammography (P=0.01),and color flow signal strength in ultrasound (P＜0.05).Conclusions:TNF alpha in breast tumor tissue was unusually high expression,and is closely related to some of the imaging features of breast tumor.

Objective: To understand the behavioral characteristics of vaccination of human papillomavirus (HPV) vaccine based on diffusion of innovation theory, and to put forward suggestions for improving the vaccination willingness. Methods: The women who had made an appointment or received at least one dose of HPV vaccine in two community health service centers with HPV vaccination qualification in Hangzhou were recruited by convenience sampling method. A questionnaire survey was conducted according to cervical cancer risk factors, HPV vaccine knowledge and the time of vaccination. The diffusion of HPV vaccination and the characteristics of vaccinees in different stages was analyzed. Results: A total of 448 women, aged from 18 to 45 years old, with a median of 30.5 years old, were investigated. The spread of HPV vaccine were divided into five stages: April 2017 and before was the initial stage, with 8 leaders, accounting for 1.79%; October 2017 to February 2018 was the slowly rising stage, with 59 early vaccinees, accounting for 13.17%; April 2018 to April 2019 was the rising stage, with 160 most early vaccinees, accounting for 35.71%; May to August 2019 was the rapidly rising stage, with 170 most late vaccinees, accounting for 37.95%; September 2019 to the end of the survey was the standstill stage, with 51 laggards, accounting for 11.38%. All of the leaders had participated in cervical cancer screening, voluntarily vaccinated, and knew the susceptible population of cervical cancer, the targeted HPV types and the suitable population of vaccine. The median age of the first pregnancy was 27 years old. Six of them had three pregnancies. The early vaccinees and the most early vaccinees were recommended by doctors or friends to get HPV vaccine. The median age of first pregnancy was 26 and 28 years old, respectively. Their other characteristics were similar to those of the leaders. The characteristics of most late vaccinees were similar to those of the laggards. They were were mainly students who were recommended by their friends to receive HPV vaccination, did not participate in cervical cancer screening, did not know the susceptible population of cervical cancer, the targeted HPV types and the suitable population of vaccine. Conclusions People with high risk of HPV exposure, awareness of cervical cancer and screening are more likely to receive HPV vaccination at the early stage. It is necessary to strengthen the health education of HPV vaccination, and make full use of the influence of doctors and early vaccinees, which is helpful to promote the early vaccination of HPV vaccine among the right age population.

Objective To investigate the effects of electromagnetic irradiation on activation of protein kinase C(PKC)and phosphorylation of glutamate receptor 2(GluR2)in rat cerebellum.Methods Sixty male Wistar rats were divided randomly into a control group and an electromagnetic exposure group(including 5 subgroups ob- served at different time points after the irradiation,eg.0 hour,3 hours,12 hours,24 hours and 72 hours after irradi- ation),with 10 rats in each group.All the rats in the exposure group were exposed to 90 mW/cm2 electromagnetic ir- radiation for 20 minutes,their rectal temperature was detected immediately after irradiation and the specific absorption rate(SAR)value was calculated,activation of PKC was detected with improved TaKai method,the level of cerebellar GluR2 expression and phosphorylation(ser880)was detected by using Western blot.Results Immediately(0 hour)after exposure,the rectal temperature of rats increased 2.99℃,SAR value was 8.66 W/kg.When compared to the control group,it was found that there was no significant difference between the exposure group and the control group with regard to all the parameters at 3,12,24 and 72 hours after exposure,except that the cerebellar PKC acti- vation and GluR2(ser 880)phosphorylation decreased significantly immediately after irradiation.Conclusion The electromagnetic irradiation has injurious effects on cerebellar signal pathway of for motor learning.