Objective To investigate the role of inhibitor of DNA binding-2 (ID2) in pulmonary vasculature formation in early lung development. Method CRISPR/Cas9 system was used to edit ID2 in human embryonic cells (hESCs) to construct ID2 knockout strains, hESCs and ID2 knockout strains were further differentiated into endothelial cells(ECs) or lung bud organoids (LBOs) .Quantitative PCR was used to detect ID2, CD31, NKX2.1 and other genes expression during differentiation, flow cytometry was performed to analyse the differentiation efficiency, Western blot was applied to check ID2, CD31 and PAX9 protein expression, 3D co-culture of ECs and LBOs was performed to compare the effect of LBOs derived from different hESCs on the tubular formation of ECs. Results ID2 can promote cells to enter the mesoderm stage during the differentiation of hESCs into ECs, but ID2 deletion leads to the formation of a large number of ECs with tubular defects. ID2 promote the differentiation of hESCs into LBOs.In the 3D co-culture system, LBOs could promote ECs tube-forming ability, while the deficiency of ID2 weakened this promotion. Conclusions In the early lung development, ID2 affects the differentiation and function of ECs and LBOs, loss of ID2 in lung bud reduces the angiogenesis of surrounded ECs.

Objective To investigate the translational regulation in human embryonic kidney (HEK)293 cell line . Methods Polysome profiling coupled with Western blot was used to verify the translational activity of HEK293 cells. RPL10A and RPS17 RNA immunoprecipitation coupled with sequencing (RIP-seq) and bioinformatic analysis were used to investigate the actively translating mRNAs. Results A total of 129 genes were found in the RNA immunoprecipitation experiments RPL10A and RPS17, among which 93.8% were protein-coding genes and enriched in RNA splicing, metabolism, MAPK signal pathway, chromatin localization and RNA transport. Conclusions A variety of molecules involved in HEK293 cells translation are obtained by RPL10A and RPS17 RIP-seq analysis.

Objective To establish an inducible CRISPR library screening method in human embryonic stem cells (hESC), and to screen the cis-acting elements involed in maintaining the pluripotency of hESC. Methods The inducible Cas9 expression elements were inserted into the hESC genome, in order to construct an inducible Cas9 stable expression strain (iCas9-hESC). Analyze the published chromatin interaction information in hESC to identify the cis-acting elements that may be involved in the regulation of pluripotency genes, and use this to construct a double gRNA CRISPR knockout library. The CIRSPR library was used to infect iCas9-hESC at low multiplicity of infection, then to induce Cas9 expression, and perform gene editing on cells. Using the expression level of OCT4 as a standard to measure pluripotency, the cells after gene editing were stained with OCT4 and analyzed by flow cytometry to determine whether the knockout of cis-elements would affect the pluripotency of hESC. Results The iCas9-hESC was successfully constructed, and the cells still maintained a pluripotent state. Construct a CRISPR knockout library of cis-elements and successfully infect the target cells. Flow cytometric analysis of the expression level of OCT4 was performed on the cells after the library infection, and it was found that there were indeed some populations with decreased pluripotency in the cells after gene editing. Conclusions A feasible scheme for screening cis-acting elements involved in maintaining pluripotency in hESC through CRISPR library is established.

Objective To explore whether ADP ribose polymerase 14(PARP14) is related to depression and whether it affects the inflammatory response of microglia. Methods Three kinds of mice models of depression were established to test whether the expression level of PARP14 is changed in depression-related brain areas by using real-time quantitative PCR and Western-bolt, including chronic restraint model(CRS), chronic unpredictable mild stress model(CUMS), lipopolysaccharide(LPS) model. Then the effect of PARP14 overexpression and knockdown on inflammatory response was detected in immortalized microglia cell line BV2. Results The expression level of PARP14 was increased in the hippocampal tissues of all three kinds of depression models, and the expression level of PARP14 in the CRS model group was correlated with the expression level of inflammatory response factors in the tissues. The expression level of PARP14 was also increased after LPS stimulation of mouse immortalized microglia cell line BV2.At the same time, BV2 cells with overexpression of PARP14 expressed more pro-inflammatory cytokines in response to LPS stimulation, while those with knockdown PARP14 or treatment with PARP14 inhibitor showed the opposite. Conclusion This study showed that the expression level of PARP14 in the hippocampus of depression model mice were increased, which may increase the level of neuroinflammation and aggravate depression-like phenotype by exacerbating the inflammatory response of microglia cells to LPS.

Objective Establishing and verifying a method for labelling neural progenitor cells (NPCs) with carboxyfluorescein ester (CFSE) fluorescent probe. Methods CFSE was injected into the lateral ventricle of embryonic mice to label ventricular zone (VZ) cells. The tissue was collected at 1 hour and 3 hours after injection to detect its specificity by immunofluorescence. Dorsal neocortex was separated at 1 hour after injection, the strong positive cells were sorted by fluorescence activated cell sorting (FACS) and the cell identity was verified by stem cell markers and neuronal markers. Results CFSE instantaneously labels (pulse labelling) cells in the ventricular zone, and the labelling window is between 3 and 6 hours. Signal of CFSE when performing long-term labelling will be attenuated. 70% of the cells sorted by CFSE are neural precursor cells, and a small portion of which are neurons. Conclusion CFSE can be used for pulse and long-term labelling in vivo. Also, CFSE can be applied to sorting NPCs, while the current sorting strategy needs to be adjusted to improve specificity.

Objective To investigate the application of CRISPR/CasRx mediated gene knockdown at the RNA level in mouse spermatogonia cell line GC1-spg. Methods The EF1a core promoter.CasRx.SV40.U6.DRs expression plasmid (CasRx-gRNA) and three fluorescent protein of EF1a.EGFP, EF1a.mCherry, EF1a.tdToamto expression plasmids were constructed by homologous recombination. Three fluorescent protein expression plasmids and CasRx-gRNA expression plasmids were transiently transfected into human embryonic kidney cell line HEK-293T, and the knockdown of exogenous gene (the fluorescent proteins，EGFP and mCherry) was used to detect by fluorescence intensity and Western blot. Additionally, the expression levels of endogenous gene (mRNA and lncRNA) were measured by q-PCR in HEK-293T cells transfected with CasRx-gRNA. The exogenous gene EGFP, mCherry and CasRx-gRNA expression plasmids were transiently transfected into the mouse spermatogonia cell line GC1, and the knockdown efficiency of exogenous gene (the fluorescent proteins) was detected by the above method. Results CasRx-gRNA and three kinds of fluorescent protein expression plasmids were constructed successfully. After CRISPR/CasRx mediated RNA interference of exogenous genes (EGFP, mCherry) and endogenous genes (mRNA, lncRNA) in HEK-293T cells, the expression level was significantly reduced (P<0.01). The CRISPR/CasRx system was verified in the mouse spermatogonia cell line GC1, which can effectively and specifically knock down exogenous genes EGFP and mCherry. Conclusions CRISPR/CasRx system can exert an effective and specific knock-down function in GC1-spg, providing a new gene silencing tool for the study of male reproduction.

Objective To explore the predictive value of detection in the efficacy of Xiaoyan decoction combined with Gemcitabine/Carboplatin (GC) chemotherapy in the treatment of advanced non-small cell lung cancer (NSCLC). Methods The clinical data of 65 patients with advanced NSCLC who were treated with Xiaoyan decoction and GC chemotherapy were analyzed retrospectively. The apparent diffusion coefficient before and after treatment (ADCpre and ADCpost) of magnetic resonance diffusion weighted imaging (MRI-DWI) and ADC change rate (ΔADC) were compared between the effective group and the ineffective group. The correlation of ADCpre and ADCpost and ΔADC with tumor regression rate and their predictive value for curative effect were analyzed. Results The effective rate of Xiaoyan decoction combined with GC chemotherapy was 58.46%. Before treatment, DWI showed high signal, ADCpre was low, and DWI was mainly equal signal and ADCpost was high after treatment. The tumor regression rate was negatively correlated with ADCpre, and positively correlated with ADCpost and ΔADC (P < 0.05). The sensitivity, specificity, accuracy and AUC of ADCpre for efficacy prediction were 71.05%, 81.48%, 75.38%, 0.789, respectively, and ADCpost was 89.47%, 66.67%, 80.00%, 0.810, and ΔADC was 81.58%, 85.19%, 83.08%, 0.842. Conclusions ADCpre, ADCpost and ΔADC are closely related to tumor regression rate in advanced NSCLC patients. MRI-DWI can predict the clinical efficacy of Xiaoyan decoction combined with GC chemotherapy.

Objective To explore the degree of radical surgery for patients with medium and high-risk gastrointestinal stromal tumors (GIST) and to find the correlation between targeted drug therapy and survival prognosis, and then to summarize the clinical diagnosis and treatment experience. Methods A retrospective analysis of the complete clinicopathological and follow-up data of intermediate and high risk GIST patients diagnosed by surgery and pathology in Shanxi Bethune Hospital from September 2012 to December 2019, and summarized the prognostic factors. Results The patients' gender, age, tumor location, rupture, degree of surgical treatment, and targeted therapy were different from those at 3 and 5 years. Tumor location, rupture, degree of surgical treatment, and targeted therapy are independent prognostic factors that affect the 3-year and 5-year OS of GIST patients. The difference between targeted therapy and 5-year OS is significant. Different degree of surgical treatment, different risk levels, and whether to use targeted drug therapy have different prognosis. Conclusions Complete surgical resection (R0) combined with targeted therapy is the main treatment to improve the prognosis of patients with intermediate- and high-risk GIST, but patients with intermediate-risk GIST who received targeted drug therapy after surgery are failed to significantly improve their overall survival.

Objective To investigate the effect and mechanism of orexin-A(OX-A) on the expression and function of glial glutamate transporter-1(GLT-1) and delayed neuronal death (DND) in hippocampus CA1 region of transient global cerebral ischemic rats. Methods The rats were divided into sham group, model group, OX-A group and OX-A+LY294002 group. The binding ability and glutamate uptake of GLT-1 in hippocampal CA1 region were detected by the [3H]-glutamate labelling method. The content of glutamate in hippocampal CA1 region was determined by the glutamate quantitative kit. The pathological changes of neurons in hippocampal CA1 region were observed by Nissol, NeuN and Fluro-Jade C staining. The expressions of GLT-1, p-PI3K and p-Akt in hippocampal CA1 region were detected by Western blot. Results OX-A promoted the binding ability and glutamate uptake of GLT-1, increased glutamate content, GLT-1 expression and PI3K/AKT signaling activity, and inhibited DND in the hippocampal CA1 region of tGCI rats (P<0.05); however, LY294002 attenuated the above therapeutic effects of OX-A (P<0.05). Conclusions OX-A can increase the expression and function of GLT-1 and reduce DND in the hippocampal CA1 region of tGCI rats by enhancing the activity of I3K/AKT signal.

Objective To explore the role of angiopoietin-like protein 3 (ANGPTL3) in the development of thoracic aortic aneurysm (TAA) and its possible mechanism. Methods The TAA patients who underwent open surgical repair in Beijing Anzhen Hospital were selected as the experimental group (n=6), and the aortic tissue in the control group was from a heart transplant donor (n=6). Another 30 3-week-old male C57BL/6J mice were randomly divided into control (control) and β-aminopropionitrile (BAPN)-induced TAA model groups (n=15 per group). The mice were taken 4 weeks later Aortic tissue. Immunohistochemical method was used to detect the expression of ANGPTL3 in human thoracic aortic aneurysm tissue, immunohistochemistry and Western blot methods were used to detect the expression of ANGPTL3, IL-6, MMP2, and MMP9 in mouse aortic tissue, and TUNEL method was used to detect apoptosis. Immunofluorescence co-localization was used to detect the localization of ANGPTL3 in vascular tissues. Transfect human aortic smooth muscle cells with ANGPTL3 siRNA. Observe the effect of ANGPTL3 knockdown on AngII-induced smooth muscle cell apoptosis and inflammatory response. Results In the aortic tissues of patients with thoracic aortic aneurysm and TAA mice, the expression level of ANGPTL3 protein increased significantly (P<0.05), vascular smooth muscle cell apoptosis, and the protein expressions of inflammatory factors IL-1β, MMP2, and MMP9 were all significantly increased. Increase (P<0.05). In addition, ANGPTL3 knockdown can significantly reduce AngII-induced smooth muscle cell apoptosis and inflammatory factor secretion (P<0.05). Conclusions The ANGPTL3 protein levels increased in the development of thoracic aortic aneurysm. The inhibition of ANGPTL3 protein expression can reduce the cell apoptosis and inflammation of smooth muscle cells. It suggested that targeting ANGPTL3 may inhibit the development of thoracic aortic aneurysms by inhibiting cell apoptosis and inflammation in smooth muscle.

Objective To investigate MBNL3 expression and its effect on malignant biology behavior of pancreatic cancer cells. Methods Fifty paraffin samples of pancreas cancer excised during surgery in Peking Union Medical College Hospital from 2017 to 2020 were collected. Immunohistochemistry was conducted to explore expression of MBNL3. Pancreatic cancer cells were treated with MBNL3 knocked down or overexpressed by transfection. Abilities of proliferation and invasion or migration in different pancreas cells were examined respectively by real time cellular analysis and Transwell invasion or migration tests. Abilities of colony forming in different pancreas cells were examined by plate colony test. The MBNL3 protein expression in cell lines was analyzed by Western blot. Results Overall staining score of MBNL3 proteins in pancreatic cancer tissues was 5.0±2.7, significantly higher than the adjacent tissues, which was 2.1±0.6（P<0.05）. Cell proliferation experiment showed that 72h after transfection, MBNL3 knocked down groups grew slower than control group in pancreatic cancer cells, whereas MBNL3 overexpressed groups grew faster than control group. Transwell experiments showed that 10h after transfection, invasion and migration ability of MBNL3 knocked down groups were weaker than control groups in pancreatic cancer cells （P<0.05）, meanwhile MBNL3 overexpressed groups were stronger than control groups in pancreatic cancer cells （P<0.05）. Colony forming experiments showed that colony formation number of MBNL3 knocked down groups were less than control groups in pancreatic cancer cells （P<0.05）, and the same of MBNL3 overexpressed groups were more than control group in pancreatic cancer cells（P<0.05）. Conclusions MBNL3 is highly expressed in pancreatic cancer tissue, which might take part in regulating the proliferation, invasion and migration of pancreatic cancer cells.

Objective To conduct molecular epidemiological investigations on Han and Buyi women in Libo County, Guizhou Province, and to study the genetic polymorphisms of key enzymes folate metabolism MTHFR and MTRR. Method A total of 1 342 healthy women who received perinatal health care at the People's Hospital of Libo County were studied, including 236 Han people and 1 106 Buyi people. Oral mucosal epithelial exfoliated cells were collected, genomic DNA was extracted, and MTHFR C677T, A1298C, and MTRR A66G gene polymorphisms were detected using a quantitative quantitative PCR method for statistical analysis. Result 1) The genetic polymorphism distribution of the subjects was in line with genetic balance. 2) The genotype frequencies of MTHFR 677CC, CT, and TT of Han women were 49.6%, 39.8%, and 10.6%, respectively, and that of Buyi women were 61.8%, 33.7%, and 4.4%. The genotype of MTHFR 1298AA, AC, and CC of Han women The frequencies were 62.3%, 33.1%, 4.7%, and Buyi females were 47.4%, 42.9%, and 9.7%, and both points are different (P <0.05). The genotype frequencies of MTRR 66AA, AG, and GG of Han women were 56.4%, 37.4%, and 5.9%, respectively, and that of Buyi women were 57.0%, 36.0%, and 7.1%, with no difference. 3) Han and Buyi women MTHFR C677T and A1298C had 6 combinations of two-point linkage, without CT / CC, TT / AC and TT / CC. The highest frequency of Han women was CT / AA, and the highest frequency of Buyi women was CC / AC. The haplotype constructed by the two loci had three combinations of CA, TA, and CC, and there was a complete linkage disequilibrium between the two loci. Conclusions Obtaining the population genetic characteristics of MTHFR and MTRR gene polymorphisms in Han and Buyi women in Libo County provides a basis for guiding scientific supplementation of folic acid nutrition and implementation of personalized pregnancy health care.

Objective To explore the mechanism of miR-183 regulating mitochondrial autophagy in hypoxia/reoxygenated （H/R） hippocampal neurons. Methods The mouse hippocampal neuron HT-22 cells were divided into control group, hypoxia/reoxygenation （H/R）model group (HR group), miR-183 overexpression group (miR-183 mimics group), and negative control group (miR-183 NC group) ). The hypoxia and reoxygenation model was established by the method of hypoxia 3h+reoxygenation 12 h, miR-183 mimics group and miR-183 NC group were transfected with miR-183 mimics and miR-183 NC on the basis of HR group. The survival rate of HT-22 cells in was measured by MTT method, the ultrastructure of mitochondria was observed by transmission electron microscope, and the levels of miR-183 and mitochondrial autophagy related genes Bnip3L mRNA were detected by qRT-PCR; Western blot was used to detect the expression levels of autophagy related proteins Bnip3L, beclin-1, ubiquitin and LC3 binding protein p62 (p62), microtubule related protein light chain 3 (LC3-Ⅱ)/LC3-Ⅰ. The targeting relationship between miR-183 and Bnip3L was verified by double Luciferase Report. Results Compared with control group, the autophagy of membrane structure in HT-22 cells increased in HR group and miR-183nc group, and melanin remained after autophagy, cell survival rate, miR-183 and p62 protein expression levels were all decreased (P < 0.05), the expressions of Bnip3L mRNA and protein, beclin-1 and microtubule-associated protein 1 light chain 3-Ⅱ (LC3-Ⅱ)/LC3-Ⅰ proteins were all increased (P < 0.05); In the miR-183 mimics group, except for the reduced cell survival rate, all the above indicators are opposite to the HR group and have statistical differences (P < 0.05). compared with miR-183 NC + Bnip3L-3'utr-wt group, the activity of luciferase in miR-183 mimics + Bnip3L-3'utr-wt group decreased (P < 0.05). Conclusions The over expression of miR-183 can target to inhibit the expression of Bnip3L mRNA, inhibit the mitochondrial autophagy of HT22 cells, and aggravate the damage of HT22 cells caused by H/R.

Objective To investigate the effect of proanthocyanidins(PC) on amyloid beta protein induced by Aβ25-35 in human neuroblastoma cell line SH-SY5Y and its possible mechanism. Methods SH-SY5Y cells were exposed to Aβ25-35 to establish cell injury model in vitro. PC and(or) secretory protein Dickkopf-1 (Dkk1), the endogenic inhibitor of Wnt/β-catenin signaling pathway were used for intervention. The cells were divided into control group, Aβ25-35 group, PC(different concentrations)+Aβ25-35 groups, Dkk1 group, Dkk1+ PC+Aβ25-35 group.The cell viability was evaluated by MTT assay. The expression levels of β-catenin, GSK-3β, p-GSK-3β were measured by Western blot. The secreted Aβ1-42 was detected by ELISA. Results Compared with the control group, the cell viability, the expression of β-catenin and p-GSK-3β were reduced in Aβ25-35 treated group(P<0.05), while the secreted Aβ1-42 was increased (P<0.05). PC improved the cell viability, increased the expression of β-catenin and p-GSK-3β, and reduced the secreted Aβ1-42 in SH-SY5Y cells induced by Aβ25-35 (P<0.05). After using Dkk1,the ability of PC was decreased on the enhancement of β-catenin expression, the improvement of p-GSK-3β expression, and the reduction of the secreted Aβ1-42 (P<0.05). Conclusions PC may decrease Aβ1-42 secretion in SH-SY5Y cells induced by Aβ25-35 through Wnt/β-catenin signaling pathway.

Objective To investigate the association between whole blood iron, copper and their interaction with cardiovascular diseases (CVD). Methods A cohort-based case control study was conducted. Cases were 93 males with CVD in the Kailuan cohort who participated in the follow-up in 3 hospitals in 2017-2018. During the same period, 372 males with non-CVD who were 1:4 individual matched by age and labor type were selected as controls. The concentration of blood iron and copper was determined by inductively coupled plasma mass spectrometry. Conditional logistic regression models were used to estimate the association between blood iron and copper and CVD, respectively and elastic-Net regression models to estimates interaction between the two metals and CVD. Result The whole blood iron concentration in the cases was significantly higher than the controls, while blood copper concentration was lower than the controls (both P<0.001). After adjusting for age, smoking, drinking, overweight, hypertension, diabetes, anemia, hyperlipidemia, and high sensitivity C-reactive protein, the participants with higher iron increase 3.17 times (OR=4.17, 95%CI: 2.23-7.79, P<0.001) risk of CVD than those with lower iron, and those with lower copper increase 3.26 times (OR=4.26, 95%CI: 1.73-10.45, P<0.01) risk than those with the higher copper. Adaptive Elastic-Net regression analysis showed that it under the simultaneous exposure of higher iron and lower copper, the risk of CVD with environment risk score > 0.232 was 8.96 times (OR=8.96, 95%CI: 4.47-17.95, P<0.001) than the score ≤ 0.232. Conclusions Higher iron and copper deficiency may be independent risk factors for CVD. Environment risk score could be a good indicator of simultaneous exposure of blood iron and copper, which have a better predictive value of CVD.

Objective To explore the role of annexin A1 (ANXA1) in resveratrol regulation of M2 type polarization of rat microglial cell line with oxygen glucose deprivation/reoxygenation (OGD/R) model. Methods Lentivirus LV-shANXA1 was used to silence the expression of ANXA1 in N9 cells. LV-Scramble acted as a control lentivirus. Western Blot and RT-qPCR were used to verify the efficiency of lentivirus transfection. 3 d after lentivirus transfection, the N9 cell OGD/R model was constructed. 3 d after re-oxygenation, the cells were treated with resveratrol for 24 h. The groups are as follows: OGD/R(-) group, OGD/R(+) group, OGD/R(+)+Res group, OGD/R(+)+Res+LV-Scramble group, OGD/R(+)+Res+LV-shANXA1. RT-qPCR and Western Blot were used to detect the mRNA and/or protein content of ANXA1, CD11b, tumor necrosis factor (TNF-α), Interleukin-1β (IL-1β), arginase-1 (Arg-1) and Interleukin-10 (IL-10). Immunofluorescence was applied to observe the distribution of ANXA1 in cells. Results The content of ANXA1 mRNA and protein in OGD/R(+)+Res group was significantly higher than that in OGD/R(+) group. The ANXA1 protein in the OGD/R(+) group was mainly distributed in the nucleus, while the ANXA1 protein in the OGD/R(+)+Res group was mainly distributed in the cytoplasm. After effectively inhibiting the expression of ANXA1, the OGD/R(+)+Res+LV-shANXA1 group ANXA1 is mainly located in the nucleus. Compared with the OGD/R(+)+Res group, the CD11b protein content in the OGD/R(+)+Res+LV-shANXA1 group was significantly increased (P<0.01), and the TNF-α and IL-1β mRNA levels were significantly increased (P<0.001), while Arg-1 and IL-10 mRNA levels were significantly reduced. Conclusions Resveratrol promotes the polarization of rat microglial cell line wih OGD/R model to M2 type via upregulating of ANXA1 expression.

Objective To compare the effect of spinal anesthesia and general anesthesia on postoperative sleep quality in elderly patients undergoing hip surgery. Methods In this prospective cohort study, patients with hip fracture, aged ≥65 years were divided into 2 group according to the anesthesia method: combined spinal and epidural anesthesia (CSEA) group and general anesthesia (GA) group (n=69 each) from November 2019 to August 2020. All patients received the same perioperative pain management. Subjective sleep quality was assessed by the sleep diaries recording sleep quality, total sleep time, and number of awakenings. The objective sleep quality was assessed by sleep tracker recording total sleep time (TST), rapid-eye-movement (REM) time, light sleep time and deep sleep time. Results Compared with CSEA group, the sleep quality score and deep sleep time were significant lower (P<0.05), and the number of awakenings (P<0.05) was significant higher in GA group on the night of surgery. Conclusions Compared with CSEA group, sleep disruption is more severe in the GA group at the night on the surgery day in patients with hip fracture.

Objective To explore the clinical value of fiberoptic ductoscopy in the diagnosis and treatment of pathological nipple discharge.Methods A total of 313 patients with pathological nipple discharge from October 2018 to June 2019 in the Beast Surgery Department of Tongzhou Maternal＆Child Health Hospital of Beijing were retrospectively analyzed. Results Fiberoptic ductoscopy was performed in 313 patients with pathological nipple discharge, 124 cases were space occupying lesions and underwent surgical treatment. Postoperative pathology revealed 6 cases of ductal carcinoma in situ, 1 case of lobular carcinoma in situ, 5 cases of atypical hyperplasia, 5 cases of intraductal papillomatosis, 6 cases of multiple intraductal papilloma, 38 cases of intraductal papilloma, 4 cases of fibroadenoma, 9 cases of breast adenosis and 50 cases of hyperplasia of mammary glands. Conclusion Fiberoptic ductoscopy can directly diagnose the etiology of pathological nipple discharge. The guide wire localization surgery guided by fiberoptic ductoscopy can more accurately remove the intraductal lesions, and fiberoptic ductoscopy can detect early breast cancer with only nipple discharge.

Objective To compare the advantages and disadvantages of full scan acquisition (FSA) mode and data dependent acquisition (DDA) mode for untargeted metabolomics analysis. Methods Qualitative and quantitative results of two acquisition methods were compared by using a standard mixture of 9 metabolites and human urine sample. Results In the analysis of standard sample, the metabolite abundance in FSA mode was about 2.36 times higher than that in DDA mode (P<0.05), the CV of metabolites abundance ranged from 2.14% to 11.20% in DDA mode and from 0.81% to 6.35% in FSA mode. In the analysis of urine sample, there was no statistical difference in the number of spectra and metabolite feature between FSA mode and DDA mode. When injecting 1, 2 and 4 μL, the number of metabolite identified by FSA mode was 9.5 %, 1.9 % and 10 % higher than those identified by DDA mode, respectively. And the median CV of metabolite abundance analyzed by FSA（7.1 %、3.0 % and 2.6 %）was significantly lower than that by DDA（10.3 %, 7.0% and 6.5%）. When injecting 1, 2 and 4 μL，with the increase of abundance, the median CV of metabolite abundance in DDA mode was 1.3 ~ 1.7，1.7 ~ 1.8 and 1.4 ~ 1.9 times higher than that in FSA mode (P<0.05), and with the increase of peak width, the median CV of DDA mode was 1.5 ~ 1.8, 1.7 ~ 2.1 and 2.0 ~ 2.2 times higher than that of FSA mode (P<0.05). Conclusions FSA mode requires high system stability, it is suitable for short-time small-scale sample analysis; DDA mode has lower requirement for system stability, and is convenient and time-saving, which can be used for long-time large-scale sample analysis.

From August 2010 and August 2020, there were 3 cases of pathologically diagnosed alveolar adenoma (AA), which is a rare benign neoplasm of the lung. The clinical manifestations and radiological characteristics on computer tomography of AA were heterogenous among these patients and lacked specificities. The difficulty in predicting the benign or malignant characteristic of the lesion lead to some confusions in clinical practices. This paper reported 2 cases of AA and reviewed the literature of AA. We analyzed its clinical, radiographical and pathological characters, hoping to provide more knowledge and improve the differential diagnosis of this disease.

Although there are many pathological mechanisms of diabetic cardiomyopathy (DCM), oxidative stress is one of the most important causes of DCM. Understanding the occurrence and development of oxidative stress in DCM is very important for the new targeted therapy of DCM. In addition, hydrogen sulfide (H2S) plays an important role in the treatment of various cardiovascular diseases. In particular, H2S can reduce DCM by regulating various ways to inhibit oxidative stress.

Atherosclerosis is a chronic inflammatory disease involving imbalance of lipid metabolism and immune disorders. A disintegrin and metalloprotease 17（ADAM17) is a classic membrane protein shedding enzyme, which promotes atherosclerosis by participating in the process of inflammatory response, angiogenesis and plaque instability. In a few cases, ADAM17 also exhibits anti-atherosclerotic effects. Therefore, paying attention to the mechanism of ADAM17 in atherosclerosis will help provide new insights into the diagnosis and treatment of atherosclerosis.

Post-transcriptional regulation of mRNA is an essential step between RNA transcription and gene translation, which includes mRNA splicing, nuclear export, translation, and decay. Post-transcriptional dysregulation in the brain is one of the causal factors of Alzheimer’s disease. Here we summarize the dysfunctional posttranscriptional regulation of APP, MAPT and many other risk genes of AD identified so far. Meanwhile, RNA N6-methyladenosine methylation is the most abundant epitranscriptomic mark on mRNA and plays a regulatory role of on almost every step in mRNA metabolism. Due to the abnormal expression of m6A regulators and participation in alternative splicing of risk genes in AD, the hypothesis proposes that post-transcriptional dys-regulation mediated by m6A may function as a novel pathogenic factor of AD.

Acidic nucleoplasmic DNA-binding protein 1 (And-1/WDHD1) as a natural chimeric protein, is characterized by containing the WD-repeat and the HMG-box families. And-1 is over-expressed in multiple tumors and involved in the whole cell cycle containing prereplication complex (Pre-RC) assembly, DNA replication, checkpoint activation, DNA repair and sister chromatid cohesion establishment. As a novel target for cell cycle regulation, And-1 can be used in the development of broad-spectrum anti-cancer drugs. This article focuses on the research on the structural characteristics, biochemical functions and potential roles of And-1 in cancer in recent years, in order to provide references for the further research of And-1 and the development of new drugs.

Objective To explore the practical effect of Harvard Medical School’s T2T (Training-to-Teachers) program for young clinical educators and improve the construction of medical discipline teaching echelon. Methods A total of 26 teachers from Chinese Academy of Medical Sciences & Peking Union Medical College Hospital were selected to participate in the T2T program. T2T Project lasted for eight months, which consists of 7 medical education module of online courses, 8 live webinars, 2 online examinations (Midterm Quiz and Final Examination), 1 Capstone Project and 2 workshops held in Peking Union Medical College and Harvard Medical School respectively. At the end of the project, the feedback results of real-name questionnaires of all participating teachers were used to evaluate the practical effect of T2T program. Results 61.5% of the 26 teachers participating in the project are women and 65.4% of them have senior professional titles. All the teachers undertake certain teaching tasks, among which 76.9% participate in undergraduate teaching at Chinese Academy of Medical Sciences & Peking Union Medical College Hospital. Program members give a high appraisal to the whole program: more than 90% of the members agree that they have learnt the advanced medical education concept (96.2%), curriculum design method (96.1%), the practical teaching methods and teaching evaluation idea (96.1%), and most of them agree that they have learnt the practical design method for teaching and scientific research (69.3%) and that the content of the course arrangement is reasonable (65.3%). Among all the program curricula, members consider that the offline workshop (73.1%) and Capstone Project (69.2%) are the most fruitful parts. After the end of the program, all teachers intend to improve the current teaching situation based on what they had learned. 80.8% of the teachers would try new practical teaching methods, and 65.4% of the teachers would write teaching articles based on the content of the program. Conclusions Generally, Harvard Medical School's T2T training program for young clinical educators has achieved good results, which is of great guiding significance for the establishment of a new Chinese medical education talent training system and the improvement of teaching level and quality.

The quality of traditional sMDT (single-center multi-disciplinary treatment team) teaching model will be affected by the center's location, medical level, medical habits, dominant disciplines, and superior technologies, which will directly affect the depth and breadth of students' knowledge of diseases such as tumors. Based on the traditional sMDT teaching, the new multi-centered MDT (mMDT) teaching model establishes cooperative teaching relationships with multiple hospitals and disciplines, co-cultivates undergraduate and graduate students, and regularly conducts mMDT teaching, which effectively avoids the defects of the traditional sMDT teaching model and meets the needs of students at different levels. Through the mMDT teaching mode, students can learn the advanced clinical thinking and techniques in different disciplines or hospitals, have a deeper understanding of the concept of comprehensive diagnosis and treatment of tumors, and improve the comprehensive quality of clinical and scientific research.

Objective: To establish an evaluation model of scientific impact of supervisors and provide reference for the construction of supervisor team. Methods: Based on the literature review, a series of evaluation index were selected; and 62 supervisors for doctor degree from a well-known hospital in China were selected. Their academic conditions were evaluated by factor analysis. Results: The evaluation of supervisor's scientific impact mainly relied on three common factors, including research output factor, research investment factor and academic works factor, and the total variance contribution of three factors reached 75.25%.There were significant differences in the scientific impact of supervisors. Totally, supervisor's research input was greater than their research output. Conclusion: Factor analysis can be used to construct the evaluation model of mentor's scientific impact. At the same time, it is of great significance to evaluate the scientific impact of supervisor for the selection of high-level mentors and the construction of supervisor team.

Objective To evaluate the online teaching of Traditional Chinese Medicine for undergraduates in Nursing College of Peking Union Medical College in 2020. Methods Totally 144 students participated in the whole process of learning were asked to fill in the questionnaire of traditional Chinese medicine (TCM) teaching by means of anonymous questionnaire, focusing on 7 parts of questions：the understanding of TCM and TCM courses, the understanding of online teaching and its comparison with offline teaching, the evaluation of students' autonomous learning ability, the comprehensive evaluation of teachers' teaching, the evaluation of teaching effect, the evaluation of examination and TCM teaching The overall evaluation of the course and the evaluation of the future application of traditional Chinese medicine were statistically analyzed. Ｒesults Totally 144 questionnaires were collected. About 90% of the students are interested in TCM and think it is necessary to study TCM. 76.39% of the students thought that the teaching platform of Tencent conference was the best. In online teaching, 34.72% of the students fully listen to the lectures, and 63.89% of the students listen to the lectures in part of the time. 48.61% think that the effect of online teaching is similar to that of offline teaching, and 22.22% of the students think it is better. More than 99% of the students think that teachers have mastered the skills of using online teaching platform, and the evaluation of teachers' teaching and teaching effect is positive. 66.67% students hope to introduce MOOC, 45.83% students hope to introduce micro class, 38.89% students hope to use flipped classroom. 55.56% of the students chose open book examination combined with usual attendance for final assessment. 39.58% of the students thought that TCM was difficult to learn, and 46.93% thought that TCM could not be self-taught. More than 50% of the students said that they would let their patients receive TCM treatment or nursing in the future, and about 45% said that they might. Conclusions It is necessary for medical students in western hospital to study traditional Chinese medicine and increase the class hours of traditional Chinese medicine courses. Online teaching puts forward higher requirements for teachers, and teaching evaluation can promote the improvement of teaching quality.

There was a gap between the level of clinical performance/research in Western China and that in Eastern China. Through the investigation and comparison of the staff composition, beds number and research publication in the Department of Otolaryngology of two affiliated hospitals of Western and Eastern medical colleges in 2020, the weakness and barriers of medical personnel in Western China were found out. The regular Journal Clubs (JCs) is an effective method which may stimulate the interest of faculty at hospital in clinical research, enhance research level and arouse the enthusiasm of paper publication with extensive view of latest medical progress and improve the quality of healthcare service.