BackgroundHuman retinal gliocytes play an important role in proliferative diseases,which are the basis of in vitro studies.Researchers have cultured human retinal gliocytes in the past.In our study,we found that the cells we cultured presented a unique shape different from those by other researchers.ObjectiveThis study was to design to produce a new culture and purification method for retinal gliocyte in vitro.Methods Retinal tissue was isolated from human eyeballs and digested using the two-step digestion method (2％ pancreatin and 0.133％collagenase Ⅵ) to harvest the retinal glio cytes.The cells were collected and cultured in endothelial cell-targeted nutrient culture containing 10％ fetal calf serum and supplemented with β-endothelial cell growth factor (ECGF) and sodium heparin,and the culture dishes were coated with fibronectin(FN) to promote the attachment of retinal gliocyte.During the culturing process,the gliocytes were identified by the observation of morphological characteristic and regular histological examination.The identification of the cells also was performed by immunochemistry targeting glial fibrillary acidic protein (GFAP),Vimentin,neuron specific enolase ( NSE ),S-100,CD34,and Ⅷ factor.Results Retinal gliocytes were isolated successfully from the human retina by the two-step digestion method.Primary cultured cells attached after 72 hours and achieved confluency between day 9 and 10 that were aligned petaliform in shape.Regular histological examination after H&E staining showed blue cell nuclei and light red cytoplasm.The target cells presented with strong responses for GFAP and Vimentin and no response for NSE,S-100,CD34 and Ⅷ factor.ConclusionsLarge amount of purified human retinal gliocytes can be obtained by two-step digestion and cultured in endothelial cells-targeted culture medium supplemented with β-ECGF and sodium heparin in plates coated with FN.The cultured cells expressed markers for retinal gliocytes.However,specific features of these cells remain to be further elucidated.

mPem, a homeobox gene, is expressed in a time and stage specific manner during murine ontogeny. Pem transcripts are abundant in 7- and 8-day mouse embryos, but decrease precipitously thereafter. On Day 9 they become abundant in placenta and yolk sac, persisting there until parturition. Although Pem transcripts are not detectable in most of adult tissues, they are present in reproductive system such as testis, epididymis and ovary. This indicates a important role for Pem during embryogenesis and reproductive development. To study the function of mPem protein, we used a GAL4 based yeast two-hybrid assay to screen a 7-day mouse embryo library with full-length of mPem. 3 proteins were found interacting with mPem protein. One of theses is Mdfic. We confirmed the interaction between mPem and Mdfic in yeast and in vitro. Mdfic, MyoD family inhibitor domain containing, encodes the myoD family inhibitor domain (I-mfa domain). The interaction between mPem and Mdfic suggested they maybe form the transcriptional regulator complex to regulate embryo differentiation.
Animals ; Embryo, Mammalian ; Embryonic Development ; Female ; Genes, Homeobox ; genetics ; Homeodomain Proteins ; chemistry ; metabolism ; Mice ; Pregnancy ; Protein Binding ; Transcription Factors ; chemistry ; metabolism ; Two-Hybrid System Techniques

Objective To investigate the expressions of human telomerase reverse transcriptase(hTERT) mRNA and protein in pheochromocytoma and paraganglioma and their significance as diagnostic markers in predicting the biological behaviour of these tumours.Methods Expression of hTERT mRNA was determined by in situ hybridization in 45 pheochromocytomas/paragangliomas(31 benign,7 suspected malignant and 7 malignant) and 9 normal adrenal medulla samples,hTERT protein was determined by immunohistoebemistry.Results hTERT mRNA was expressed in 5/7 malignant turnouts and 5/7 suspected malignant tumours as compared with 3/31 benign tumours(P

Objective:To study the expression of phosphates signal transducer and activator of transcription 3 (pSTAT3) and programmed death ligand-1 (PD-L1) in extranodal NK/T cell lymphomas (ENKTCL) and the relationships of pSTAT3 and PD-L1 expression with the clinicopathological characteristics and prognosis of ENKTCL.Methods:Fifty-one cases of ENKTCL diagnosed at Guangdong Provincial People′s Hospital from June 2015 to February 2019 were included in the study. The expression of pSTAT3 and PD-L1 was examined using immunohistochemistry.Results:There were 35 males and 16 females, ranging from 18 to 85 years old with a median age of 47 years. The positive rates of pSTAT3 and PD-L1 expression were 68.6% (35/51) and 76.5% (39/51), respectively. pSTAT3 expression was correlated with PD-L1 expression ( P=0.033, R=0.322), while there were no associations of pSTAT3 and PD-L1 expression with the clinicopathological characteristics of ENKTCL, including age, sex, clinical site, B symptom, Ann Arbor stage, LDH value, EBV DNA load of peripheral blood and international proliferation index score. Kaplan-Meier survival analysis showed the prognoses of the pSTAT3 and PD-L1 positive groups were slightly better than the respective negative groups, but the differences were not significantly ( P>0.05). Conclusions:pSTAT3 is highly expressed in extranodal NK/T cell lymphoma and related to the expression of PD-L1, which provides a potential target and rationale for combinations of targeted therapies and immune checkpoint blockade inhibitors in the treatment of ENKTCL.

Objective To analyze the epidemiological characteristics of imported cases of dengue fever in Guangdong province (GD) and in I-long Kong (HK) during 2004-2006 to provide evidence for further cooperation in the prevention and control programs on dengue fever in the two places.Methods Descriptive statistical analysis was performed on data obtained from dengue fever surveillance and reporting network in GD and from Centre for Health Protection,Department of Health,HK.Results Both from GD and HK 44 and 93 imported cases of Dengue fever were reported during 2004-2006.Most patients from GD acquired their infection from Singapore (13 cases),Indonesia (9 cases) or Cambodia (6 cases) while patients in HK mainly were imported from Indonesia (31 cases),the Philippines (16 cases) and Thailand (15 cases).The peak seasons of the two places were both from July to September.During the non-peak season period,the number of cases in Hong Kong was higher than that in Guangdong.Male/ female ratio was 1.2 : 1 in GD and 1.1 : 1 in HK.Age of patients in GD appeared to range from 6-80 years,with 63.6% (28/44) of them aged 20-39 years.40.9% (18/44) of the cases were engaged in business,services,housework or unemployed.Those cases in HK were between 10-72 years of age,with 63.6%(28/44) of them aged 20-39 years while 47.3%(44/93) of the patients were with the occupation of business,services and industry.More cases in GD had a onset of disease before entering the border (27 : 17) than the cases in HK (35 : 57).The average time interval between onset and diagnosis were 7 and 9 days for GD and HK respectively.Conclusion Frequent travel between Southeast Asia in summer among the working class appeared to be the main factor,causing imported cases of dengue fever in GDand HK.It is crucial to provide health education targeted at these high risk groups in order to prevent importation of dengue fever in the two areas.

Objective To investigate the lipid levels of Han ethnicity Chinese children at school-age,to provide objective data for the formulation of prevention and management strategy regarding dyslipidemia among children and adolescents.Methods 20 191 children (with 10 669 boys and 9522 girls) aged 7 to 16 years old from 6 representative geographical areas,including Beijing,Tianjin,Hangzhou,Shanghai,Chongqing and Nanning,were surveyed in a randomly selected clustered sample in China.Data on fasting blood triglyceride (TG),total cholesterol (TC),lowdensity lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) levels were measured.Non-high-density lipoprotein cholesterol (non-HDL-C) levels were calculated with data collection,entry,and collation were under the same criteria.Results (1) In the 7-16 year-old group,TG (P95) fluctuated between 1.26 mmol/L and 1.88 mmol/L,while TC (P95) was between 4.80 mmol/L and 5.46 mmol/L.LDL-C(P95) was between 2.67 mmol/L and 3.27 mmol/L while non-HDL-C (P95) was between 3.36 mmol/L and 3.91mmol/L,sugesting that age did not seem to be an affecting factor for the lipid level (P＞0.05).The level of HDL-C (P5) fluctuated bctwcen 1.08 mmol/L and 0.83 mmol/L,and the dependability analysis on HDL-C and age showed statistically significant difference (P＜0.01,r=-0.274).(2) In the 7-9 year-old group,the levels ofTG,TC,LDL-C and non-HDL-C of boys were lower but the HDL-C level was higher than in girls.However,in the 10-16 year-old group,the levels of five lipids of boys were all lower than in girls,with all the differences statistically significant (P＜0.05).(3) The levels of TG,TC,LDL-C and non-HDL-C in the obese group were significantly higher than those in non-obesity group,as HDL-C was significantly lower than in non-obese group(P＜0.01).Incidence rates of single and multiple dyslipidemia in obese group were significantly higher than in non-obese group (P＜0.01).(4) Grouped by region,the abnormal rates of TG were descending,with the ranking as North (10.4％),Midwest (9.7％) and East (8.3％),while the abnormal rates of TC were descending with the ranking as Midwest (6.0％),North (5.2％)and East (4.8％).The abnormal rates of LDL-C were descending as the ranking of North (3.1％),East (2.6％) and Midwest (0.9％),with the abnormal rates of non-HDL-C were descending as Midwest (6.5％),North(4.2％)and East (3.6％).The abnormal rates of HDL-C were descending as Midwess (14.2％),North(5.7％) and East(5.5％).All the differences in the above-said items were statistically significant (P＜0.05).(5) According to the standards of hyperlipidemia formulated by the American Academy of Pediatrics,the incidence rates of abnormal TG,TC,LDL-C,non-HDL-C,HDL-C were 9.4％,5.4％,2.2％,4.8％,8.6％ respectively.Conclusion (1) Levels of lipids were affected by many factors,but age was not one of them in children and adolescents.However,HDL-C was declining along with the increase of age,to some extent.(2)Girls had a relatively protective tendency through the increasing HDL-C level when they entered the puberty years.(3)Lipids levels in non-obese group were significantly better than the obese group.(4)The lipids levels of children and adolescents in the Eastern region of the country were better than that in the northern and mid-western areas.

OBJECTIVETo investigate the anti-HIV constituents from the root of Mirabilis jalapa.

METHODThe compounds were isolated by column chromatography on silica gel, Sephadex LH - 20, MCI-gel CHP-20P and RP-18. The structure were identified by means of NMR and MS analyses (1H-NMR, 13C-NMR, MS).

RESULTEleven compounds were isolated and identified as astragaloside II (1), astragaloside II (2), astragaloside IV (3), astragaloside VI (4), flazin (5), 4'-hydroxy-2, 3-dihydroflavone 7-beta-D-glucopyranoside (6), gingerglycolipid A (7), 3, 4-dihydroxybenzaldehyd (8), p-hydroxybenzaldehyde (9), beta-sitosterol (10) and daucosterol (11).

CONCLUSIONCompounds 1-9 were obtained from this genus for the first time.

Benzaldehydes ; analysis ; chemistry ; Carbolines ; analysis ; chemistry ; Chromatography, Gel ; Furans ; analysis ; chemistry ; Galactolipids ; analysis ; chemistry ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Mirabilis ; chemistry ; Plant Roots ; chemistry ; Sitosterols ; analysis ; chemistry

UNLABELLEDMorbidity of chronic lung disease in young children is relatively high, and could increase in the future. Pulmonary function testing is used for clinical assessment of patients with suspected or obvious pulmonary disease to assess the severity of dysfunction and to evaluate therapeutic effectiveness. In the recent few years, forced expiratory parameters assessing lung function have been measured in older children. In order to assess abnormalities of lung function in preschool patients with respiratory disorders based on changes of forced expiratory parameters, adequate reference values are needed. However, such data in healthy preschool children remain scant in the literature.

OBJECTIVEThe aim of this study was to characterize the spirometry of preschool children and establish the normal lung function prediction equations for Chinese preschool children.

METHODSA survey in 343 healthy preschool children (184 boys and 159 girls) aged 3 to 6 years (73 children aged 3 years, 96 children aged 4 years, 91 children aged 5 years and 83 children aged 6 years) was carried out in Shenzhen in 2004. Eleven flow volume tests parameters, i.e., forced vital capacity (FVC), forced expiratory volume at 0.5 second (FEV(0.5)), forced expiratory volume at 0.75 second (FEV(0.75)), forced expiratory volume at one second (FEV(1)), maximal mid expiratory flow rate (FEF(25%-75%)), peak expiratory flow (PEF), forced expiratory time (FET100%) were measured by using COSMED spirometry produced in Italy. Stepwise multiple regressions and non-linear regressions were carried out with the statistical software SPSS10.0 for Windows to identify the best predictors of lung function parameters using standing height, weight, age and gender as potential determinants.

RESULTSSpirometric tests could be successfully carried out by using imagery methods in the following percentages of children: 69.9% of 3 to 4 years old, 70.8% of 4 to 5 years old, 92.3% of 5 to 6 years old and 91.6% of 6 to 7 years old children, 77.7% of the selected population (217/279) of children performed at least two acceptable tests respectively. The average forced expiratory time (FET) was 1.61 +/- 0.52 sec (x +/- s), the 5th percentile value was 0.9 sec, 18 of 279 (6.5%) children produced a FET less than 1 second. Forced expiratory volume in 0.50 and 0.75 sec (FEV(0.5), FEV(0.75)) were thus measured necessary in preschool children. All lung volumes and flow rates increased with age, height as well as weight growth in both gender groups. The correlation among most lung function parameters was higher in height than in age and weight in boys. Whereas the correlation among most lung function parameters was higher in age than in height and weight in girls. The regression equations of lung function were established. By comparison with the equations derived from this study and from studies by Nystad, it was found that there was a difference between the predicted values.

CONCLUSIONSpirometric testing is feasible in preschool children by using imagery methods and may be useful for both clinical practice and research. The correlation among most lung function parameters was higher in height than in age and weight in boys. Whereas the correlation among most lung function parameters was higher in age than in height and weight in girls.

Age Factors ; Body Height ; Body Weight ; Child ; Child, Preschool ; Feasibility Studies ; Female ; Forced Expiratory Volume ; Humans ; Male ; Peak Expiratory Flow Rate ; Reference Values ; Regression Analysis ; Respiratory Function Tests ; methods ; Spirometry ; statistics & numerical data ; Vital Capacity

OBJECTIVETo uncover the transmission patterns of the HIV epidemic in Dehong prefecture.

METHODSThe reviewed case reports, data of sentinel surveillance, testing and special survey were analyzed by SAS 8.0 program. The transmission patterns were modeled by utilizing data including sizes of the whole population and the high risk groups, high risk behavior data from 1989 to 2007, and the population index such as sex ratio and fertility rate.

RESULTSIn 2005, case reports showed the proportion of people infected with HIV through sexual contact was 39.1%, and 46.9% in 2006. Among 1636 cases reported between January 1 to September 20, 2007, the proportion of people infected with HIV through sexual contact was 52%. From 1989 to 2007, the proportion of HIV infection among drug users was declining, while HIV infection through sexual contact was rising after standardizing the population tested/surveyed. The Asian Epidemic Model has shown that the proportions of incident HIV infections through sexual transmissions were 50.6%, 52.3% and 52.7% respectively from 2005 to 2007. Correspondingly, the proportions of incident cases by injecting drug user were 48.9%, 47.2% and 46.7% respectively during this period. Moreover, the Workbook method has shown that, among adults living with HIV in 2007, 50.3% were infected through injecting drugs and 48.4% through unsafe sexual activity.

CONCLUSIONThe rapid rise in HIV infections through injecting drug in Dehong prefecture has been initially curbed. HIV epidemic has already witnessed a change from predominantly through drug injecting-related activity to an almost equally fuelled epidemic by sexual and drug-related transmission.

Acquired Immunodeficiency Syndrome ; epidemiology ; transmission ; China ; epidemiology ; Humans ; Models, Statistical ; Risk Factors ; Social Problems

Objective To investigate the influence of dezocine on spinal cord glial fibrillary acidic protein (GFAP) in rats with chronic constriction injury (CCI) of sciatic nerve as well as its protective effect on sciatic nerve.Methods Thirty two SD rats were randomly divided into sham-operation group,model group,experimental-L,-H groups (with 2.5,10 mg · kg-1 dezocine),eight SD rats in each group.The SD rats in experimental groups were treated with dezocine by intraperitoneal injection once every day from the beginning to seven days after operation;but the SD rats in sham-operation group and model group were treated with normal saline.Seven days after operation,the changes of pathological morphological of sciatic nerve in the four groups were detected.And the expression of GFAP in the four groups was detected by immunofluorescence histochemistry.Results After continuous injection of dezocine for 7 days,the sciatic nerve fibers in experimental-L group rats were swelling,the number of sheath cells decreased,and some of them were dissolved.In experimental-H group,there were a few normal myelin in rats,the swelling of nerve fibers is reduced compared with those in model group,but the number of sheath cells decreased compared with those in sham group,axons can be seen in some myelin sheaths.The expression of glial fibrillary acidic protein of spinal cord in sham-operation group,model group,experimental-L,-H groups were 1.03 ± 0.28,2.08 ±0.55,1.61 ±0.14,1.25 ±0.45.Compared with sham group,the difference in model group was significant (P ＜0.001);compared with model group,the difference in experimental-L,-H groups was significant(P ＜ 0.05,P ＜ 0.01).Conclusion Dezocine can inhibit the neuropathic pathologic pain.The mechanism maybe related to the decrease of the expression of GFAP in spinal cord,the activation of astrocytes,and the relief of the edema of sciatic nerve.