Objective To explore the experiment condition and method for the application of in vitro in vasive Transwell chamber and to observe muscarinicreceptor stimulant and muscarinicreceptor antagonist's influence to cholangiocarcinoma's invasiveness.Methods Two hundred microliter cell suspension of various concentrations(0.5×105/mL,1.0×105/mL,1.5×105/mL and 2.0×105/mL)was added into the upper chamber of the Transwell chamber,and the cells were allowed to penetrate the matrigel for 12,18,24and 48 hours respectively.The numbers was gotten as the invasive cells on the under surface of the membrane.After optimal cell concentration and time were gotten,pilocarpine of various concentrations(0 mmol/L,0.1 mmol/L,0.3 mmol/L and 0.5 mmoL/L)was added into the upper chamber of the Transwell chamber,then the cells on the matrigel were stained and counted.So did the cells when atropine of various concentrations(0.01 mmol/L,0.01 mmol/L,0.05 mmoVL and 0.1 mmol/L)were added into the upper chamher of the Transwell chamber in according to pilocarpine of various concentrations(0 mmol/L,0.3 mmol/L,0.3 mmol/L and 0.3mmol/L).Results With the increase of the time and cell concentrations，the cells couts that penetrated the matrigel increased，while the increase tended to he stable when the culture time exceeded 36 hous and the cell concentration Was over 1.0×105/mL.By adding pilocarpine,there were significant differences between the control and experimental groups(P＜0.05)，but there were no significant differences in experimental groups with various concentrations.There were no significant differences in blank group and experimental groups with atropine added(P＞0.05).When added pilocarpine and atropine,there were significant differences between blank and experimental groups(P＜0.05)，but there were no significant differences in experimental groups with various concentrations.Conclusions Thirty-six hours as invasive time,and one cell concentration 1.0 × 105/mL were optimal to test invasion abilities of cholangiocarcingma cells to different medicines or reagents.There is the possibility that museariniereceptor exists in cholangiocarcinoma cells，and may play an important role in cholangiocarcinoma's invasiveness and metastasis.

BACKGROUND:Previous studies have confirmed that bone marrow mesenchymal stem cells in vitro can promote hepatic stel ate cellapoptosis and inhibit its activity, in which the mechanism of action remains unknown. OBJECTIVE:To screen out apoptosis-related genes during hepatic stel ate cellapoptosis regulated by bone marrow mesenchymal stem cells using gene chip technology. METHODS:Purified human bone marrow mesenchymal stem cells were seeded in 6-wel Transwel plate and cocultured with hepatic stel ate cells. Cultured human bone marrow mesenchymal stem cells alone served as control group, and cultured for 72 hours. The alterations in apoptosis-related genes were analyzed between culture alone group and coculture group using gene chip technology. The genes strongly associated with regulation of hepatic stel ate cells were selected. RESULTS AND CONCLUSION:By the functional classification of second-generation SABiosciences Gene chips, apoptotic gene screening found that after coculture, significantly upregulated genes in bone marrow mesenchymal stem cells contained:AKT1, PIK3R2, DAPK1, DHCR24, NOTCH2 and BDNF. Combined with previous findings, we hypothesized that NOTCH may play a key role in the regulation of hepatic stel ate cells by bone marrow mesenchymal stem cells.

Objective:To evaluate the changes in topological properties of brain functional network after induction of general anesthesia in the patients with glioma.Methods:Twenty-two patients scheduled for elective intracranial glioma resection were selected.Resting-state functional magnetic resonance imaging was performed during wakefulness and general anesthesia with endotracheal intubation in patients with glioma. Large-scale functional brain networks of each patient were constructed based on 123 regions of interest in non-surgical hemisphere. Global properties (local efficiency, clustering parameter, shortest path length, global efficiency, small world) and nodal properties (nodal degree, nodal efficiency, and between centrality) in brain functional networks were then compared between wakefulness and general anesthesia.Results:Eighteen patients were finally enrolled. Compared with the status during wakefulness, the local efficiency and clustering parameter on non-surgical side significantly decreased ( P<0.05), no significant change was found in the shortest path length and global efficiency ( P>0.05), and small world was greater than 1 throughout the entire density range; the nodal degree, nodal efficiency and between centrality of nodes located in the medial/mesal regions, such as the medial prefrontal cortex, posterior cingulate gyrus/precuneus, medial temporal lobe, anterior cingulate gyrus, thalamus, amygdala, were significantly reduced ( P<0.05); however, these node parameters increased significantly in the lateral brain regions ( P<0.05) except for the primary auditory and somatosensory cortex, which also decreased significantly after induction of general anesthesia( P<0.05). Conclusions:The functional segregation of brain functional network is widely inhibited after induction of general anesthesia, but the functional integration is still retained. The lateral brain regions show no anticorrelation with the medial brain region during general anesthesia.

Objective:To study the efficiency and difference of the artificial intelligence (AI) system based on fundus-reading in community and hospital scenarios in screening/diagnosing diabetic retinopathy (DR) among aged population, and further evaluate its application value.Methods:A combination of retrospective and prospective study. The clinical data of 1 608 elderly patients with diabetes were continuously treated in Henan Eye Hospital & Henan Eye Institute from July 2018 to March 2021, were collected. Among them, there were 659 males and 949 females; median age was 64 years old. From December 2018 to April 2019, 496 elderly diabetes patients were prospectively recruited in the community. Among them, there were 202 males and 294 female; median age was 62 years old. An ophthalmologist or a trained endocrinologist performed a non-mydriatic fundus color photographic examination in both eyes, and a 45° frontal radiograph was taken with the central fovea as the central posterior pole. The AI system was developed based on the deep learning YOLO source code, AI system based on the deep learning algorithm was applied in final diagnosis reporting by the"AI+manual-check" method. The diagnosis of DR were classified into 0-4 stage. The 2-4 stage patients were classified into referral DR group.Results:A total of 1 989 cases (94.5%, 1 989/2 104) were read by AI, of which 437 (88.1%, 437/496) and 1 552 (96.5%, 1 552/1 608) from the community and hospital, respectively. The reading rate of AI films from community sources was lower than that from hospital sources, and the difference was statistically significant ( χ2=51.612, P<0.001). The main reasons for poor image quality in the community were small pupil (47.1%, 24/51), cataract (19.6%, 10/51), and cataract combined with small pupil (21.6%, 11/51). The total negative rate of DR was 62.4% (1 241/1 989); among them, the community and hospital sources were 84.2% and 56.3%, respectively, and the AI diagnosis negative rate of community source was higher than that of hospital, and the difference was statistically significant ( χ2=113.108, P<0.001). AI diagnosis required referral to DR 20.2% (401/1 989). Among them, community and hospital sources were 6.4% and 24.0%, respectively. The rate of referral for DR for AI diagnosis from community sources was lower than that of hospitals, and the difference was statistically significant ( χ2=65.655, P<0.001). There was a statistically significant difference in the composition ratio of patients with different stages of DR diagnosed by AI from different sources ( χ2=13.435, P=0.001). Among them, community-derived patients were mainly DR without referral (52.2%, 36/69); hospital-derived patients were mainly DR requiring referral (54.9%, 373/679), and the detection rate of treated DR was higher (14.3%). The first rank of the order of the fundus lesions number automatically identified by AI was drusen (68.4%) and intraretinal hemorrhage (48.5%) in the communities and hospitals respectively. Conclusions:It is more suitable for early and negative DR screening for its high non-referral DR detection rate in the community. Whilst referral DR were mainly found in hospital scenario.

Objective:To investigate the perioperative alterations and management of coagulation function in patients of massive blood transfusion during retroperitoneal tumor (RT)resection.Methods:Fourty-seven RT patients at Peking University International Hospital from Jan 2016 to Dec 2021 undergoing resection with massive blood transfusion more than 20 U within 24 h were reviewed for coagulation function before and after surgery.Results:Intraoperative bleeding was 3 000-25 800 ml, 10 patients had blood loss ≥10 000 ml. During the operation, (25.3±9.9) U of red blood cells were transfused, (2 720±1 369) ml plasma transfused, and (2.4±3.3) U platelets were transfused in 6 patients. Fourty-five patients received intraoperative albumin of (79.5±46.5) g; All 47 patients received fibrinogen of (2.3±1.3) g; Prothrombin complex was given in 45 patients (1 205±807) U. Preoperative hemoglobin was statistically different compared to postoperatively and days 1, 3 and 5 ( W=1 790, P<0.001; W=1 672, P<0.001; W=1 704, P<0.001; W=1 486, P=0.004);As with platelets, the difference was also statistically significant compared to postoperative days 1, 3, and 5 ( W=2 153, P<0.001; W=2 092, P<0.001; W=1 732, P<0.001); Preoperative albumin was different compared to postoperative days 1 and 3 ( W=1 568, P<0.001; W=1 578, P<0.001,); Preoperative fibrinogen was different compared to postoperative day 1 ( W=1 964, P<0.001). PT and APTT were prolonged on postoperative days 1 and 3 ( W=628, P<0.001, W=804, P=0.023) ( W=661, P<0.001, W=796, P=0.02). Patient's preoperative fibrin degradation products and D-dimer were above the normal value and were higher on postoperative days 3 and 5 ( W=498, P<0.001, W=345, P<0.001). Conclusions:Coagulation disorders occur perioperatively in patients with massive transfusion while undergoing surgery for RT.The implementation of ratiional transfusion strategy and close postoperative survey and management of coagulation dysfunction help avoid the coagulation related morbidities.

Objective:To assess the prognostic value of the collateral status and clot burden score based on four-dimensional computed tomography angiography(4D CTA)in anteriorcir culation is chemics troke patients with large ischemic core after endovascular treatment.Methods:Clinical and imaging data of 36 anterior circulation ischemic stroke patients with large infarct core (infarct core≥50.0 ml) after endovascular treatment at our institution from March 2016 to September 2020 were retrospectively reviewed. According to the modified Rankin Scale (mRS) score, patients were divided into the good outcome (mRS score 0-2) and poor outcome (mRS score 3-6) groups. Mann-Whitney U and Fisher tests were used to compare the 4D CTA collateral circulation score, clot burden score, and baseline clinical data between the good and poor outcome groups. Multivariate logistic regression was used to analyze the risk factors associated with the poor outcome (mRS score 3-6) and mortality in patients with large infarct core stroke. Finally, based on the 90-day outcome, a ROC curve was used to obtain the cut-off values for poor prognosis (mRS 3-6) and death, respectively. Results:Ten patients (27.8%) had good outcome and 26 (72.2%) had poor outcome. The patients in the poor outcome group had older median age, higher blood glucose, lower 4D CTA collateral circulation score, lower clot burden score, larger infarct core volume, and higher hemorrhagic transformation and brain hernia (all P<0.05). Multivariate logistic regression showed that the poor collateral circulation score on 4D CTA( OR=0.18, 95% CI: 0.03-0.99, P<0.05)and clot burden score( OR=0.64, 95% CI: 0.44-0.93, P<0.05) were independent predictors of the poor prognosis. The ROC curves revealed that the cut-off value of infarct core for distinguishing between good prognosis and poor prognosis was 63.7 ml, while that for distinguishing between survival and death was 130.3 ml. Conclusions:Endovascular treatment may improve the prognosis of patients with large infarct core of anterior circulation is chemic stroke if the patients have good 4D CTA collateral circulation score and high clot burden score.

Myocardial fibrosis （MF） is a common pathological manifestation of various heart diseases. Due to the non-renewable nature of myocardial cells， the occurrence of MF represents irreversible damage to the myocardium. Previous studies have suggested that fibroblast-mediated collagen deposition is the main mechanism of MF. Recent studies have found that there is an immune regulation mechanism in the heart itself， and macrophage activation/polarization plays an important role in MF. With the deepening of traditional Chinese medicine research， scholars have found that traditional Chinese medicine can interfere with MF by regulating the renin-angiotensin-aldosterone system （RAAS） system and the inflammatory process， repairing the extracellular matrix， managing oxidative stress， and maintaining the balance of autophagy. This process is closely related to the activation and M1/M2 polarization of macrophages. Throughout the MF process， macrophage activation is beneficial， but excessive activation will be harmful. In the early stage of MF， appropriate M1 macrophage polarization is conducive to activating immunity and removing harmful substances. In the middle and late stages of MF， appropriate M2 macrophage polarization is conducive to remodeling the damaged myocardium. If macrophage activation is excessive/insufficient， or the balance of M1/M2 macrophage polarization is broken， the effect changes from improvement to destruction. Traditional Chinese medicines that regulate the activation/polarization of macrophages have the effects of replenishing Qi and nourishing Yin， as well as regulating Qi and activating blood， but there are also some heat-clearing， dampness-drying， and detoxification products. Therefore， the occurrence of MF may be caused by Qi and Yin deficiency， damp heat accumulation， and Qi stagnation and blood stasis. By summarizing the biological processes involved in macrophage activation/polarization in MF， this paper expounded on the research progress of traditional Chinese medicine in regulating macrophage activation and M1/M2 polarization from different angles to improve MF， so as to provide a reference for the treatment of MF with traditional Chinese medicine.

Animals ; SARS-CoV-2 ; Antibodies, Neutralizing ; Macaca mulatta ; COVID-19/prevention & control* ; Antibodies, Viral ; Vaccines