In order to explore the platform role and practicability of network teaching in the teaching and training of clinical blood transfusion. By building an "Internet+APP" teaching and training management platform, this research develops personalized teaching and training courses and assessment plans for different groups such as interns, trainees and on-the-job staff in the blood transfusion department, so as to achieve time-saving and high-efficient training results. The results showed that the interns' assessment scores were all up to standard, with more than 90 points accounting for 66% and 80-90 points accounting for 34%. The assessment scores of the trainees and on-the-job personnel were above 90 points, which showed that they had significant improvement of their professional level. The teaching resources of this teaching mode are stable, centered on the trainees, free from traditional time and space constraints, high-efficient, time-saving, and easy to accept.

Various c-mesenchymal-to-epithelial transition (c-MET) inhibitors are effective in the treatment of non-small cell lung cancer; however, the inevitable drug resistance remains a challenge, limiting their clinical efficacy. Therefore, novel strategies targeting c-MET are urgently required. Herein, through rational structure optimization, we obtained novel exceptionally potent and orally active c-MET proteolysis targeting chimeras (PROTACs) namely D10 and D15 based on thalidomide and tepotinib. D10 and D15 inhibited cell growth with low nanomolar IC₅₀ values and achieved picomolar DC₅₀ values and >99% of maximum degradation (D_max) in EBC-1 and Hs746T cells. Mechanistically, D10 and D15 dramatically induced cell apoptosis, G1 cell cycle arrest and inhibited cell migration and invasion. Notably, intraperitoneal administration of D10 and D15 significantly inhibited tumor growth in the EBC-1 xenograft model and oral administration of D15 induced approximately complete tumor suppression in the Hs746T xenograft model with well-tolerated dose-schedules. Furthermore, D10 and D15 exerted significant anti-tumor effect in cells with c-MET^Y1230H and c-MET^D1228N mutations, which are resistant to tepotinib in clinic. These findings demonstrated that D10 and D15 could serve as candidates for the treatment of tumors with MET alterations.

【Objective】 To investigate the profile of Rh blood group antigen in pregnant women and hemolytic disease of the newborn (HDN) in Qingdao area. 【Methods】 10 597 pregnant women admitted in our hospital during October 2016 to February 2020 were selected and the ABO, Rh blood group system antigen (D, C, c, E, e) and the irregular antibody were detected, and positive antibody was further identified. The irregular antibody of Rh blood group in pregnant women was statistically analyzed according to the history of blood transfusion and pregnancy. Twelve HDN cases were studied, and the results of ABO, Rh blood group antigen and irregular antibody, antibody property identification, HDN test and blood routine test were retrospectively analyzed. 【Results】 Among 10 513 cases of Rh-positive pregnant women, the common phenotype was CCee>CcEe>Ccee>ccEE>ccEe; among 84 cases of Rh-negative pregnant wome, the common phenotype was ccee>Ccee> CCee> ccEE>ccEe. The positive rate of irregular antibody was 1.06% (112/10597) in 10 597 pregnant women, of which the Rh antibody was the highest, rated at 56.25% (63/112). For 64 pregnant women with positive antibodies, antibodies against Rh system were different from those against other systems when stratified by the history of blood transfusion (P<0.05) and pregnancy (P<0.05). Twelve neonates were diagnosed with Rh-HDN, with IgG anti-E in 6 cases, IgG anti-D 3, IgG anti-cE 1, IgG anti-C 1and IgG anti-c 1. Among them, 3 were seriously ill and treated with blood exchange. 【Conclusion】 As two-child policy was implemented, the incidence of Rh HDN had increased. ABO, RhD, C, c, E and e matched transfusion should be administered for women at childbearing age. Meanwhile, clinical termination of delivery was recommended for pregnant women, who probably develop Rh-HDN and are with critical situation. Rh phenotype matched fresh blood should be prepared, which has great clinical significance for rescuing newborns.

【Objective】 To investigate the distribution characteristics of Rh blood group antigen phenotypes, haplotypes and irregular antibodies between patients in our hospital and local blood donors, so as to ensure safe and effective blood transfusion and improve the rationality and scientificity of clinical blood transfusion. 【Methods】 A total of 113 326 blood samples, from hospitalized patients in our hospital and local blood donors from October 2015 to March 2020, were subjected to Rh antigen typing and irregular antibody detection. The frequency of Rh phenotypes, haplotypes, and irregular antibodies were retrospectively analyzed and calculated. Chi square test was used to compare the data among different population groups. Rh antigen typing and irregular antibody detection were completed using the automatic blood group analyzer. 【Results】 The prevalence of negative RhD was 0.36% (408/113 326). The most prevalent Rh phenotype was DCCee [40.69%(46 112/ 113 326)] followed by DCcEe [36.82%(41 727/ 113 326)]. Anti-E was the most common irregular antibody, accounting for [0.26%(295/ 113 326)], and DCe [62.51%(70 840/ 113 326)] was the most common haplotype. The most common Rh phenotypes and haplotypes in Caucasians in Germany, North Indian and North African were DCcee, DCCee and Dccee, while DCe, DCe and Dce, respectively. 【Conclusion】 The distribution characteristics of Rh phenotypes, haplotypes and irregular antibodies of patients in our hospital and local blood donors were in line with the distribution characteristics of the population in northern China. Corresponding plans concerning blood storage and collection, as well as the establishment of Rh blood type registry should be carried to effectively ensure the safety, rationality and accuracy of clinical blood transfusion.

Objective To construct engineered corneal epithelium from embryonic stem cells (ESCs) using Rock inhibitor combined with hypoxia-normoxia culture condition. Methods Human ESC line H1 was induced to differentiate into epithelial-like cells by addition of retinoic acid (RA) and bone morphogenetic protein 4(BMP4) in the differentiation medium under the adherent culture condition. The ESCs derived epithelial-like cells were expanded in the mixed medium of SHEM and KSFM with the mixture ratio of 1 : 2 with or without Rock inhibitor Y27632. The H1 derived epithelial-like cells were seeded on the denuded ammonic membrane to construct engineered corneal epithelium under hypoxia,normoxia and hypoxia-normoxia culture conditions,respectively. The inducted effect of ESCs into epithelial-like cells,the expansion ability of the epithelial-like cells and the characteristics of the constructed engineered corneal epithelium were evaluated by morphological observation, real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and immunofluorescence technology. Results Compared with the control group,the relative expressions of ESCs marker Oct4 mRNA, Notch signaling pathway related factors Notch1 and Jagged1 mRNA,and Wnt signaling pathways related factors c-myc and Cyclin D1 mRNA were significantly reduced, and the relative expressions of cutaneous ectoderm markers p63 and K18 mRNA were significantly increased at day 8 after induction in the induced group,with significant differences between them (t =14.63,20.15,93.50,11.60, 19.30,18.44,22.63;all at P<0.05). Compared with the without Y27632 group,the relative expressions of p63 and K14 mRNA,Notch signal pathway receptor Notch1 and Jagged1 mRNA were significantly increased,and Wnt signaling pathways downstream targeted gene c-myc and CylinD1 mRNA were significantly decreased at day 8 after induction in the Y27632 group,with significant differences between them (t =20.29,59.22,2.90,39.59,5.32,10.14;all at P<0.05),and the relative expression of K18 mRNA in the two groups was not significantly changed(t=1.38,P>0.05). The ESCs derived epithelium and constructed under hypoxia-normoxia culture condition showed more obvious stratification and tighter cell arrangement in comparison with those cells cultured in consistent hypoxia culture condition or normoxia culture condition. Epithelial markers Pan-CK and K18 as well as epithelial progenitor cell markers p63 and K14 expressed in the whole cell layers of the ESCs derived epithelium constructed under hypoxia-normoxia culture condition. Conclusions The addition of Y27632 enhances the proliferation ability of H1 derived epithelial cells and actives Notch signaling pathway and inhibits Wnt signaling pathway. The culture and construction in the expansion medium with Y27632 under the hypoxia-normoxia culture condition can promote the stratification of H1 derived engineered corneal epithelium.

Sleep deficiency is a common public health problem associated with many diseases, such as obesity and cardiovascular disease. In this study, we established a sleep deprivation (SD) mouse model using a ‘stick over water’ method and observed the effect of sleep deficiency on ocular surface health. We found that SD decreased aqueous tear secretion; increased corneal epithelial cell defects, corneal sensitivity, and apoptosis; and induced squamous metaplasia of the corneal epithelium. These pathological changes mimic the typical features of dry eye. However, there was no obvious corneal inflammation and conjunctival goblet cell change after SD for 10 days. Meanwhile, lacrimal gland hypertrophy along with abnormal lipid metabolites, secretory proteins and free amino-acid profiles became apparent as the SD duration increased. Furthermore, the ocular surface changes induced by SD for 10 days were largely reversed after 14 days of rest. We conclude that SD compromises lacrimal system function and induces dry eye. These findings will benefit the clinical diagnosis and treatment of sleep-disorder-related ocular surface diseases.

Objective To investigate the regional cerebral stimulation after central analgesics nasal spray and its mechanism with pharmacological functional magnetic resonance imaging (phfMRI). Methods Eighteen healthy right-handed volunteers participated. Butorphanol tartrate nasal spray was used as the experiment agent. Ethological experiment was carried out to record the participants' subjective feeling and the onset time of the analgesics, followed by the functional MRI (fMRI) scan two weeks later. Block design was adopted. Two phases of fMRI scan were performed at 7 min and 25 min after the nasal spray, respectively. Participants were also given pain stimulation in the dorsum of hand during the fMRI scanning. The data were post-processed with Matlab 6.5 and SPM 2. Results ①Onset time of butorphanol tartrate was 15-35 min after nasal spray administration, which was consistent with its concentration-time curve. ②After nasal spray, activations were observed in the cerebral cortex, including frontal lobe (orbitofrontal gyrus, medial frontal gyrus, superior frontal gyrus), temporal lobe (insula, middle temporal gyrus, inferior temporal gyrus), parietal lobe (precuneal gyrus), limbic system (anterior cingulate gyrus, middle cingulate gyrus, hippocampus, parahippocampal gyrus);subcortical region (globus pallidus) and cerebellum (6-9 of cerebellar cortex, cerebellar peduncle, vermis). ③The number and activation intensity of the second phase were more obvious than those of the first phase (P<0.01). Conclusion The feasibility of phfMRI study on cerebral stimulation and the mechanism of nasal spray is demonstrated. The study of butorphanol tartrate further validates the main distribution of opioid receptors in the central nervous system and the possible mechanism of central analgesia.

OBJECTIVETo observe the effect of the extract of Ginkgo biloba (EGb761) and the components isolated from the extract named ginkgolide B (GB) against damage of glutamate in pretreatment modes so that determine their application value and approach.

METHODBased on glutamate-induced excitotoxicity to primary cultures from neonatal Sprague-Dawley (SD) rat hippocampal neuron, our experiment utilized trypan blue, TUNEL and LDH to study the effect of EGb761 and GB on neuron in different doses pretreatment modes, as well as to compare with the NMDA receptor uncompetitive antagonist-MK-801.

RESULTEGb761 and GB can recrease cell viability, reduce apoptosis rate and decrease LDH leakage in different degree and depended on dose in certain range. The maximal protection was achieved at a concentration of 100 mg x L(-1), 100 micromol x L(-1), but inferior to MK-801 (10 micromol x L(-1)). The protective effect of GB is superior to EGb761.

CONCLUSIONTreatment with EGb761 and GB could protect the neurons against glutamate-induced injury. The maximal protection of GB was achieved by pretreatment is superior to EGb761, so its precautionanary intervention to high-risk population could have more value.

Animals ; Animals, Newborn ; Cells, Cultured ; Dizocilpine Maleate ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Ginkgo biloba ; chemistry ; Ginkgolides ; chemistry ; pharmacology ; In Situ Nick-End Labeling ; L-Lactate Dehydrogenase ; metabolism ; Lactones ; chemistry ; pharmacology ; Neurons ; drug effects ; Neuroprotective Agents ; chemistry ; pharmacology ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective:To explore more reliable standards for identifying vestibular hair cells of saccule and utricle prepared in studies with patch clamp technique under inverted phase contrast microscope. Method:The length and width of two types hair cells were measured besides the length of cilia,and all datas were analyzed statistically.Result:The width and length of cilia of two types hair cells in saccule and utricle from guinea pig were similar. The length of type Ⅰ was longer than that of type Ⅱ,SO the ratio between length and width was larger and the ratio of the length between cilia and cell body was small.Conclnsion:Two type'S hair cells of saccule and utricle from guinea pig may be distinguished through the ratio of cell body's length and width even the ratio of the length between cilia and cell body,besides the standards before.

OBJECTIVE: To explore more reliable standards for identifying vestibular hair cells of saccule and utricle prepared in studies with patch clamp technique under inverted phase contrast microscope. METHOD: The length and width of two type's hair cell's were measured besides the length of cilia, and all datas were analyzed statistically. RESULT: The width and length of cilia of two types hair cells in saccule and utricle from guinea pig were similar. The length of type I was longer than that of type II, so the ratio between length and width was larger and the ratio of the length between cilia and cell body was small. CONCLUSION Two type's hair cells of saccule and utricle from guinea pig may be distinguished through the ratio of cell body's length and width even the ratio of the length between cilia and cell body, besides the standards before.
Animals ; Cell Shape ; Guinea Pigs ; Hair Cells, Vestibular ; cytology ; Microscopy, Phase-Contrast ; Patch-Clamp Techniques ; Saccule and Utricle ; cytology