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.

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

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 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

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.

Objective:To investigate the effect of traditional Chinese medicine enema retention time on patients with chronic renal failure under the intervention of guan yuan point in patients with TDP lamp irradiation.Methods:By convenience sampling, 65 patients with chronic renal failure who were admitted to the Chinese medicine department of a Class Ⅲ Grade A hospital in Hubei Province from 1st January 2019 to 1st November 2019 and needed Chinese medicine retention enema treatment were selected as the research subjects. 65 patients with chronic renal failure were randomly divided into observation group (32 cases) and control group (33 cases) by random table. The control group received routine enema operation, and the experimental group received TDP light irradiation at the guan yuan point on this basis. After two consecutive courses of treatment, the effective rate of enema, blood urea nitrogen and the decreased level of creatinine, and nursing satisfaction rate in the two groups were compared.Results:The total effective rate of Chinese medicine enema in the observation group was 81.82% (27/33) , and the total effective rate in the control group was 6.25% (2/32) , the difference between the two groups was statistically significant ( P<0.05) . After the intervention, the blood creatinine and blood urea nitrogen levels in the observation group were lower than those in the control group, and the differences between the two groups were no statistically significant ( P>0.05) . The total satisfaction rate of patients in the observation group was 93.94% (31/33) , which was higher than 71.88% (23/32) in the control group, the difference between the two groups was statistically significant ( P<0.05) . Conclusions:After TDP light irradiation on guan yuan point, the retention time of traditional Chinese medicine enema patients was prolonged, which promoted the absorption of traditional Chinese medicine, improved the therapeutic effect of traditional Chinese medicine enema, and delayed the course of disease.

Objective To study the influence of nasal irrigations on the postoperative curative effect of patients with chronic sinusitis.Methods Total of 357 patients who suffered from chronic sinusitis received transnasal endoscopic surgery from 2008 to 2010 with 184 patients selecting nasal irrigations for the following treatments.Patients were followed up at two weeks,six weeks and four months after the surgeries via telephone.The symptoms of nasal obstruction,nasal discharge and headache were recorded and SNOT-20 was used to measure the quality of life.Results Symptoms of patients were improved,but improvement in nasal irrigations group was better than that in non-nasal irrigations group.Four months later,the average symptom score of nasal obstruction in nasal irrigations group was [0(0,1 )] which was significantly lower than [ 0( 0,1 )] in non-nasal irrigations group ( Z =- 3.176,P ＜ 0.05).The average symptom score of nasal discharge in nasal irrigations group was [0(0,2)] which was significantly lower than [0(0,2)] in non-nasal irrigations group (Z =- 4.403,P ＜ 0.05 ).The average score of headache was [ 0 ( 0,1 ) ] in nasal irrigations group and [ 0 (0,2) ] in non-nasal irrigations group,and the difference was statistically significant ( Z =- 4.539,P ＜ 0.05 ).Conclusions Nasal irrigations can improve symptoms of patients with chronic sinusitis and reduce the rectrrence.