Effects of different culture systems on the culture of prepuberal buffalo (Bubalus bubalis) spermatogonial stem cell-like cells in vitro

Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⁺ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.

Effects of different culture systems on the culture of prepuberal buffalo (Bubalus bubalis) spermatogonial stem cell-like cells in vitro

Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⁺ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.

Effects of different culture systems on the culture of prepuberal buffalo (Bubalus bubalis) spermatogonial stem cell-like cells in vitro

Intervention of auricular point sticking on perioperative psychological stress in patients with anorectal diseases / 中国针灸

OBJECTIVE@#To explore and quantify the intervention effect of auricular point sticking on perioperative psychological stress in patients with anorectal diseases.@*METHODS@#Eighty patients who underwent anorectal surgery were randomly divided into an observation group (40 cases) and a control group (40 cases). The routine preoperative guidance, preoperative visits, and informed of the postoperative condition were received in the control group. On the basis of the treatment in the control group, auricular point sticking was immediately applied at Shenmen (TF), Shen (CO), Wei (CO), Gan (CO), Pi (CO), Pizhixia (AT), E (AT), Nie (AT) and Zhen (AT) in the observation group.The patients were pressed by themselves, 3 to 5 min per point each time, 5 times a day, and the contralateral auricular points were replaced every 2 or 3 days until 1 week after surgery. The Hamilton anxiety scale (HAMA), Hamilton depression scale (HAMD), and Pittsburgh sleep quality index (PSQI) scores were compared between the two groups before and 7 days after surgery.@*RESULTS@#There was no significant difference in the total HAMA scores between after and before surgery in the observation group (>0.05). The total HAMA score in the control group was higher than that before surgery (0.05). The scores of somatic anxiety factor in the two groups were higher than those before surgery (0.05). The total score of PSQI in the two groups was lower than that before surgery (<0.05), and the total score of PSQI in the observation group was lower than that in the control group (<0.05).@*CONCLUSION@#Auricular point sticking can effectively improve some psychological stress problems during perioperative period in patients with anorectal diseases.

Resolution revolution:advances and prospects of cryo-EM in structural biology / 复旦学报（医学版）

Cryo electron microscopy (cryo-EM) is one of the most important methods in structural biology.In the past five years,cryo-EM has milestone breakthrough due to revolutionary advances in hardware and methodology.The resolution has been pushed to as high as 1.8 （A),which significantly extended the research scope of this method.Single particle reconstruction method has become one of the most exciting fields of structural biology and related subjects.Here we will briefly introduce the history of cryo-EM,recent revolutionary breakthrough,and the facility at Fudan University.

Retrospect of Institute of Biomedical Sciences of Fudan University on epigenetics research / 复旦学报（医学版）

Epigenetics is the study of heritable traits that are not involved in the changes in DNA sequences.It has been widely recognized that epigenetics plays multiple regulatory roles in diverse cellular activities and is associated with the occurrence and development of a variety of diseases.Following the human genome project (HGP),discoveries in the field of epigenetics reveal the complexity and accuracy of genetic regulation.Institute of Biomedical Sciences (IBS),which is affiliated with Fudan University Shanghai Medical School,has been attaching great importance to "Medical Epigenetics" since the establishment of IBS.In the past 12 years,a number of influential discoveries in medical epigenetics have been made,raising China's status in the field of epigenetics and contributing greatly to the disciplinary construction and talent cultivation of biomedical-related disciplines in Fudan University.In this review,a list of accomplishments in epigenetics in IBS were summarized.

Clinical analysis of interleukin-6 level in tear and serum of dry eye patient / 中华实验眼科杂志

Background Mechanism of dry eye disease has not been established clearly.Some increasing evidences showed that inflammation plays an important role in pathogenesis and development of dry eye disease.However,the association of interleukin-6 (IL-6) with dry eye has not been clarified.Objective This study was to assess the correlation of IL-6 levels in tear and serum of patient with dry eye disease and clinical symptom.Methods Twenty patients with Sj（o）gren syndrome (SS) and 20 cases with non-Sj（o）gren syndrome (NSS) were enrolled from Second Affiliated Hospital of Nanjing Medical University,and 20 healthy volunteers were recruited as the normal controls.Symptom questionnaires were self-answered and multiple dry eye-related clinical tests were performed,including tear film breakup time (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescence staining.IL-6 concentrations in serum and tear were detected by enzyme-linked immunosorbent assay (ELASA).The correlation between IL-6 level and tear film and ocular surface parameters was analyzed by Spearman rank correlation.Written informed consent was obtained from each subject initial of this trial.Results The scores of symptom questionnaires were 18.50 (11.75,23.00),23.00 (19.00,32.00) and 2.00 (0.25,4.00) in the SS group,NSS group and the normal control group,and significantly increasing values were seen in the SS group and NSS group compared with the normal control group (P=0.00,0.00).BUT values in the SS group and NSS group were 2.50 (2.00,3.88) seconds and 3.50 (2.13,4.00) seconds and were lower than 15.00 (13.13,17.00) seconds in the normal control group (P=0.00,0.00).S Ⅰ t values in the SS group,NSS group and the normal control group were 2.00 (1.50,2.88),4.00 (3.00,5.75) and 19.00 (18.00,25.00)mm,showing significant differences between the SS group and the normal control group (P =0.00),the SS group and the NSS group (P =0.00) or the NSS group and the normal control group (P=0.00).The marked elevation of keratoepithelioplasty score was seen between the SS group and the normal control group (5.75 (4.00,7.75) vs.0.00 (0.00,0.75) (P =0.00) or the NSS group and the normal control group (4.50(3.63,6.00) vs.0.00(0.00,0.75)) (P=0.00).Concentrations of IL-6 in serum and tear in the SS group,NSS group were (131.47±21.04) μg/L and (77.81 ± 15.68) μg/L and were significantly higher than (31.62±8.57) μg/L of the normal control group,and those in the tear were (33.44±8.01) μg/L,(18.78 ±5.73) μg/L and (8.77 ± 3.43)μg/L,respectively in the three groups,with a significant reduce in the normal control group (all P=0.00).tL-6 levels in serum and tear were positively correlated with the score of symptom questionnaires or keratoepithelioplasty and negatively correlated with BUT and S Ⅰ t (all P =0.00).Conclusions IL-6 levels in the tear and serum of the dry eye patient are associated with the severity of the disease.IL-6 levels in serum and tear may be objective,sensitive and reliable indicators of dry eye.

The role of human papillomavirus in the pathogenesis of pterygium / 中华实验眼科杂志

Background The pathogenesis of pterygium has been in the study.Relative molecular biology study showed that pterygium is a tumor-like lesion,and based on overseas literatures,human papillomavirus (HPV) is positively expressed in 100％ patients with pterygium in some region.However,if this result is suitable for Chinese patients is unclear.Objective This study was to identify the role of human HPV in the pathogenesis of pterygia in Wuxi area.Methods Forty-eight pterygium specimens including 7 recurrent pterygia and 41 primary pterygia were collected during the operation,and these patients were from Wuxi area.Two cervical carcinoma specimens and 2 conjunctiva specimens from normal donors were obtained as positive control and negative control respectively.The fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect HPV DNA of specimens.Results The amplified curves of HPV 6,11 of pterygium specimens,cervical carcinoma specimens and normal specimens were all below the positive quality control curve,but the amplified curves of HPV16,18 were above the quality control curve in cervical carcinoma specimens; while those of pterygium specimens and normal conjunctival specimens were all below the quality control curve.HPV16/18 was identified in 2 cervical carcinoma specimens,but no HPV6/11 was detected in 2 cervical carcinoma specimens.However,HPV DNA expression in primary and recurrent pterygias were absent.Conclusions According to these results,HPV is not the primary cause for the pathogenesis of pterygium in Wuxi region.