Astrocytes are the largest glial population in the mammalian brain. However, we have a minimal understanding of astrocyte development, especially fate specification in different regions of the brain. Through lineage tracing of the progenitors of the third ventricle (3V) wall via in-utero electroporation in the embryonic mouse brain, we show the fate specification and migration pattern of astrocytes derived from radial glia along the 3V wall. Unexpectedly, radial glia located in different regions along the 3V wall of the diencephalon produce distinct cell types: radial glia in the upper region produce astrocytes and those in the lower region produce neurons in the diencephalon. With genetic fate mapping analysis, we reveal that the first population of astrocytes appears along the zona incerta in the diencephalon. Astrogenesis occurs at an early time point in the dorsal region relative to that in the ventral region of the developing diencephalon. With transcriptomic analysis of the region-specific 3V wall and lateral ventricle (LV) wall, we identified cohorts of differentially-expressed genes in the dorsal 3V wall compared to the ventral 3V wall and LV wall that may regulate astrogenesis in the dorsal diencephalon. Together, these results demonstrate that the generation of astrocytes shows a spatiotemporal pattern in the developing mouse diencephalon.
Mice ; Animals ; Astrocytes ; Neuroglia/physiology* ; Diencephalon ; Brain ; Neurons ; Mammals

Objective To investigate the effect of Gegen Qinlian Decoction on liver energy metabolism and free fatty acid(FFA)in rats with insulin resistance(IR).Methods IR rat model was established by feeding 60%fat high-fat diet for 13 consecutive weeks.SD rats were randomly divided into normal group,model group,Rosiglitazone(5 mg·kg-1)group and Gegen Qinlian Decoction low-,medium-and high-dose groups(1.65,4.96,14.86 g·kg-1),with 6 rats in each group.Intragastric administration was given once a day,continuous administration intervention lasted for 16 weeks.Determination of IR-related indicators:serum fasting insulin(FINS),fasting blood glucose(FPG),calculate the IR index;HPLC method was established for the determination of adenosine triphosphate(ATP),adenosine diphosphate(ADP)and adenosine monophosphate(AMP)in rat liver tissue;ELISA was used to determine the content of FFA in rat liver tissue.The contents of serum total cholesterol(TC)and triglyceride(TG)were detected by automatic biochemical analyzer.The pathological changes of liver tissue were observed by HE staining.Results(1)After the model replication,compared with the normal group,the FINS,FPG levels and IR index of the model group were significantly increased(P＜0.05).(2)Compared with the normal group,the levels of FINS,FPG and IR index in the model group were significantly increased(P＜0.01),the contents of ATP,ADP and AMP in liver tissue were significantly decreased(P＜0.01),the content of FFA was significantly increased(P＜0.01),and the levels of TC and TG in serum were significantly increased(P＜0.05).Liver cells arranged in disorder,fatty degeneration,and there are a large number of lipid droplets.Compared with the model group,the FINS level and IR index of rats in each administration group were significantly decreased(P＜0.05,P＜0.01),and the FPG level of rats in the Rosiglitazone group was significantly decreased(P＜0.05).The contents of ADP and AMP in liver tissue of rats in each administration group were significantly increased(P＜0.05,P＜0.01),and the contents of ATP in liver tissue of rats in low-,medium-and high dose-groups of Gegen Qinlian Decoction were significantly increased(P＜0.05,P＜0.01).The content of FFA in liver tissue of rats in Rosiglitazone group and Gegen Qinlian Decoction low-and high-dose groups was significantly decreased(P＜0.01).The serum TC level of rats in the low-and high-dose groups of Gegen Qinlian Decoction was significantly decreased(P＜0.05,P＜0.01),and the serum TG level of rats in the Rosiglitazone group and the low-dose group of Gegen Qinlian Decoction was significantly decreased(P＜0.05,P＜0.01).The steatosis of hepatocytes in rats of each administration group was alleviated to varying degrees,and the lipid droplets were reduced,and the pathological damage was improved.Conclusion Gegen Qinlian Decoction may improve liver lipid metabolism disorder and restore lipid and energy balance by regulating energy metabolism and reducing FFA level,thus improving IR.

Objective:To investigate the influence of Kruppel-like factor 16 (KLF16) on the proliferation and migration of pancreatic cancer cells.Methods:Immunohistochemical images of KLF16 were collected from 171 pancreatic cancer tissues and their matched paracarcinoma normal pancreas tissues and 8 pancreatic cancer tissues only in GEPIA database. The expression of KLF16 protein was detected by immunohistochemical imaging software. The protein and mRNA expressions of pancreatic cancer cell lines AsPC-1 and MIA PaCa-2 KLF16 were detected by Western blot and quantitative fluorescence PCR. By knockdown or exogenous overexpression of KLF16, the two cells were divided into blank control group (NC group), negative control group (siRNA-NC group), downexpression KLF16 group (siKLF16 group), overexpression control group (OE-NC group) and ovexpression KLF16-OE group (KLF16-OE group). CCK-8 assay, colony formation assay and transwell chamber were used to detect cell proliferation and migration.Results:The KLF16 protein expression level (4.02±1.26 vs 1.73±1.07) and positive expression rate (91.6% vs 13.5%) in pancreatic cancer tissues were significantly higher than those in paracancer normal pancreas tissues, with statistical significance ( P<0.05). After downregulating KLF16 expression and culturing for 24, 48, 72, and 96 hours, the A450 values of both AsPC-1 (0.19±0.02 vs 0.23±0.03, 0.24±0.06 vs 0.36±0.06, 0.45±0.09 vs 0.78±0.10, 0.69±0.04 vs 0.88±0.07) and MIA PaCa-2 cells (0.20±0.03 vs 0.22±0.02, 0.29±0.05 vs 0.31±0.04, 0.47±0.06 vs 0.78±0.10, 0.71±0.02 vs 0.90±0.07) and colony counts [(36±4.32) per well vs (118.51±10.01) per well, (13.6±2.62) per well vs (83.1±9.11) per well], and the number of migrated cells [(16.67±2.05) vs (46.67±5.91), (19.67±1.69) vs (55±4.89)] all decreased significantly. However, after up-regulating the expression of KLF16 and culturing for 24, 48, 72 and 96 h, the A450 value of both AsPC-1 (0.21±0.05 vs 0.20±0.04, 0.48±0.03 vs 0.31±0.04, 0.91±0.09 vs 0.72±0.03, 1.28±0.10 vs 1.05±0.02) and MIA PaCa-2 cells (0.20±0.01 vs 0.19±0.05, 0.44±0.03 vs 0.30±0.04, 0.89±0.06 vs 0.72±0.03, 1.19±0.05 vs 1.01±0.10), and the number of cell colonies [(189±6.37)/per hole vs (108±9.62)/ per hole, (141±12.56)/ per hole vs (80.69±10.32)/ per hole]], migration cell numbers [(79±4.89) per hole vs (50.33±4.11) per hole, (79.66±3.85) per hole vs (51±4.08) per hole] all increased significantly. Conclusions:KLF16 is highly expressed in pancreatic cancer. The up-regulated expression of KLF16 in pancreatic cancer cell lines can promote the proliferation and migration of pancreatic cancer cells.

【Objective】 To analyze the clinical features of patients with infected pancreatic necrosis (IPN) complicated with fungal infection so as to identify possible risk factors for death. 【Methods】 We analyzed the clinical data of patients with IPN admitted to Xuanwu Hospital Capital Medical University from January 1, 2017 to December 31, 2021. According to the results of pancreatic necrotic tissue and drainage fluid culture, the patients were divided into the group with fungal infection and the group without fungal infection. The baseline data, clinical features and outcomes of the two groups were compared, and the risk factors for death in patients with fungal infection were analyzed. 【Results】 We included a total of 214 patients in the study, of whom 49 patients in the fungal infection group had wider necrotic involvement, lower hematopoietic volume, and higher blood glucose at admission. Patients with fungal infection had a higher proportion of multidrug-resistant bacteria (MDRB), and hospital and ICU stay as well as parenteral nutrition duration were also longer. In the group of patients with fungal infection, the proportion of patients undergoing surgery did not increase (P>0.05), but the proportion of patients with perioperative organ failure and death was higher (P<0.05). Candida albicans (44.8%) was the most common fungus detected, followed by Candida parapsilosis (28.6%) and Candida tropicalis (8.2%). Logistic regression analysis showed that MDRB infection (OR=1.37, 95% CI:1.02-1.83), fungemia (OR=1.53, 95% CI:1.06-2.23), hyperglycemia (OR=1.65, 95% CI:1.28-2.10), new organ failure (OR=1.65, 95% CI:1.19-2.29) and bleeding complications (OR=1.64, 95% CI:1.28-2.10) after surgery were risk factors for death in patients with fungal infection. 【Conclusion】 Fungal infection increases mortality in patients with IPN and the incidence of new organ failure after surgery. Attention to fungemia, MDRB infection, hyperglycemia, organ failure and postoperative bleeding can help reduce the risk of death.

H 1-antihistamines are widely used in the treatment of various allergic diseases, but there are still many challenges in the safe and rational use of H 1-antihistamines in pediatrics, and there is a lack of guidance on the prescription review of H 1-antihistamines for children.In this paper, suggestions are put forward from the indications, dosage, route of administration, pathophysiological characteristics of children with individual difference and drug interactions, so as to provide reference for clinicians and pharmacists.

Objective:To study the clinical features and risk factors of death in patients with infected pancreatic necrosis (IPN) caused by multidrug-resistant bacteria (MDRB).Methods:The clinical data of 219 IPN patients who were managed at the Department of General Surgery of Xuanwu Hospital, Capital Medical University from January 1, 2016 to December 31, 2021 were retrospectively analyzed. There were 142 males, and 77 females, with a median age [ M( Q1, Q3)] of 51(38, 62) years old. Based on the pre-sence or absence of MDRB infection, these patients were divided into the MDRB-infected group ( n=117) and the non-MDRB-infected group ( n=102). Clinical features and outcomes were compared between the two groups, and the risk factors resulting in death in patients with MDRB infection were analyzed. Logistic regression analysis was used to determine the risk factors for poor outcomes in patients with MDRB. Results:There were significant differences in etiologies, distribution characteristics of necrosis and degrees of pancreatic necrosis between the two groups (all P<0.05). When compared with the non-MDRB-infected group, the CT severity index, the levels of procalcitonin and interleukin-6 were significantly higher in the MDRB group on admission, while the hematocrit was significantly lower (all P<0.05). Furthermore, when compared with the non-MDRB infection group, patients with MDRB infection were significantly more likely to have fungal infections [37.6%(44/117) vs. 21.6%(22/102)] and extrapancial infections [75.2%(88/117) vs. 58.8%(60/102)], more patients underwent surgery [89.7%(105/117) vs. 67.6%(69/102)], and more surgical procedures were performed [3(2, 4) times vs. 2(1, 3) times], with a higher incidence of postoperative complications [36.2%(38/117) vs. 18.8%(13/102)], an increase in a new-onset organ failure after surgery [37.1%(39/117) vs. 21.7%(15/102)], a higher in-hospital mortality rate [25.6%(30/117) vs. 10.8%(11/102)], longer hospitalization [39(28, 67) d vs. 29(18, 35) d] and ICU stays [22(10, 42) d vs. 11(6, 18) d], and a longer need for parenteral nutrition [19(9, 37) d vs. 15(7, 25) d, all P<0.05]. On multivariate regression analysis, the risk factor for death in the MDRB-infected group was co-fungal infection ( OR=1.199, 95% CI: 1.025-1.402). On the other hand, receiving therapy containing tigacycline ( OR=0.831, 95% CI: 0.715-0.965) and minimally invasive surgery ( OR=0.698, 95% CI: 0.562-0.868) reduced the risk of death in the MDRB-infected group (all P<0.05). Conclusions:IPN patients with MDRB infection had higher levels of inflammation, more serious pancreatic necrosis, longer treatment time, and increased need for surgical treatment. Measures involving fungal infection control and the use of tigacyclin and minimally invasive surgery reduced the risks of death in patients with MDRB infection.

Objective: To study the effect of stem cell factor (SCF) on the angiogenic ability of cocultured dental pulp stem cells (DPSCs) and human umbilical vein endothelial cells (HUVECs). Methods : This study has been reviewed and approved by the Ethics Committee. The experiment was split into the HUVECs, SCF+HUVECs, DPSCs+HUVECs, and SCF+DPSCs+HUVECs groups. A mixture of SCF and culture medium was used to prepare a mixed culture medium with an SCF concentration of 100 ng/mL. In vitro coculture of DPSCs and HUVECs was performed at a 1∶5 ratio. CCK-8 proliferation assay was used to observe the proliferative capacity of cells in each group on days 1, 3, 5, and 7. Wound healing and Transwell migration assays were used to detect the effect of SCF on cell migration under either direct or indirect coculture conditions, respectively. In vitro angiogenesis experiments were performed to detect the angiogenic capacity of the cells in each group. The vascular endothelial growth factor A (VEGFA) concentration in the cell culture supernatant was detected using ELISAs, and the protein expression levels of CD31, CD34, and VEGFA were detected using Western blot analysis. Results : Wound healing and Transwell migration experiments showed that SCF significantly promoted the migration of cocultured DPSCs and HUVECs (P<0.05). The in vitro angiogenesis experiment showed that the number of branches and the total length of branches of tubular structures in the SCF+DPSCs+HUVECs group were significantly greater than those of the other groups (P<0.05), and the expression levels of the vascular-related proteins CD31, CD34, and VEGFA in this group were greater (P<0.01). Conclusion SCF can enhance the migration and in vitro angiogenesis of cocultured DPSCs and HUVECs.

Objective:To analyze the effects of infectious complications [infected pancreatic necrosis (IPN) and extra-pancreatic infection (EPI)] on the outcomes of patients with severe acute pancreatitis (SAP), and evaluate the differences in infection time, infection site and infecting species between SAP patients with infections complications.Methods:The clinical data of 66 SAP patients with combined infectious complications admitted to Xuanwu Hospital, Capital Medical University from January 2014 to December 2020 were retrospectively analyzed, and SAP patients were divided into IPN group ( n=7), EPI group ( n=14) and co-infection (EPI+ IPN) group ( n=45) according to the type of infection. Whether the study data conformed to a normal distribution was assessed by the Shapiro-Wilk test, normally distributed measures were expressed as mean ± standard deviation ( ± s), and ANOVA was used for comparison between groups; skewed measures were expressed as median (interquartile range) [ M ( Q1, Q3)], and the rank-sum test was used for comparison between groups. Bonferroni correction was used for multiple group comparisons ( P value significance level reduced to 0.017). Quantitative data were compared between groups using the χ2 test or Fisher's exact probability method. Results:There were no statistical differences between the three groups in terms of baseline data at admission (gender, age, etiology, modified CTSI score, degree of pancreatic necrosis, and number of organ failure) ( P>0.05), patients in the EPI group were referred earlier than the other two groups ( P<0.05). In clinical treatment, patients in the IPN group and co-infection group required multiple minimally invasive interventions compared with those in the EPI group ( P<0.05), and the number of patients requiring combined nutritional support, length of intensive care unit stay, and total length of hospital stay were higher in the co-infection group than in the other two groups ( P<0.05). In addition, 360 strains of pathogenic bacteria were cultured in this study, with Gram-negative bacteria being the most common, and patients with SAP were more likely to have EPI in the early stage of disease onset, with bacteremia and respiratory tract infections in the early stage (≤14 d), and bacteremia, urinary tract infections, and catheter-associated infections in the late stage (>14 d). Conclusions:Among patients with SAP, patients in the co-infection group had higher surgical intervention, nutritional support and length of hospital stay than those in the single infection group. It is advisable to prioritize EPI in SAP patients with suspected infections, and the common infectious strains in SAP patients are still predominantly Gram-negative bacteria, and clinicians need to adjust the treatment plan in a timely manner according to the changes in patients′ conditions.

ObjectiveBased on ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF/MS）， the changes of endogenous markers in rat plasma at the different stage， namely modeling and administration of Shenling Baizhusan （SLBZS）， and the mechanism of SLBZS in the treatment of ulcerative colitis （UC） were studied. MethodIn the modeling stage， rats were randomly divided into normal group， spleen deficiency with dampness retention-UC （SDDR-UC） and pure-UC （P-UC） model group. In the administration stage， SLBZS was given to the above two different model groups. After modeling and administration， rat plasma was collected and determined by UPLC-Q-TOF/MS. The mobile phase was 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （in positive ion mode：0-2 min， 99%A； 2-9 min， 99%-73%A； 9-10 min， 73%-44%A； 10-13 min， 44%-38%A； 13-19 min， 38%-28%A； 19-21 min， 28%-2%A； 21-23 min， 2%A； 23-25 min， 2%-10%A； 25-27 min， 10%-99%A； in negative ion mode：0-2 min， 85%A； 2-3 min， 85%-65%A； 3-5.5 min， 65%-44%A； 5.5-8 min， 44%-25%A； 8-10 min， 25%-2%A； 10-16 min， 2%-85%A）. The electrospray ionization （ESI） temperature was 120 ℃ under the positive and negative ion modes， and the acquisition range was 50-1 000. Partial least squares-discriminant analysis （PLS-DA） was used to analyze the changes of endogenous metabolites in the above two different model rats from the different stage. MetaboAnalyst 5.0 was used to analyze the metabolic pathways of these identified metabolites. ResultSixteen potential biomarkers were screened and identified in the modeling stage， among which 11 potential biomarkers were common in the two model rats， which mainly affected the primary bile acid biosynthesis pathway. Twenty-three potential biomarkers were screened and identified during the administration stage， among which 3 potential biomarkers were shared by the two model rats， and SDDR-UC and P-UC model rats had 11 and 9 potential biomarkers， respectively. It mainly affected 6 pathways such as purine metabolism， pentose phosphate pathway， pyrimidine metabolism， retinol metabolism， primary bile acid biosynthesis and steroid hormone synthesis. ConclusionThe primary bile acid biosynthesis pathway appears in the different stage of modeling and administration of UC， showing a dynamic change process. The therapeutic effect of SLBZS on SDDR-UC rats may be related to inhibiting the expression of nuclear transcription factor -κB （NF-κB） signaling pathway， activating farnesoid X receptor （FXR） and enhancing the expression of cytochrome P450.

Objective:To investigate the computed tomography (CT) features and diagnosis and treatment of emphysema pancreatitis (EP).Methods:The retrospective and descriptive study was conducted. The clinical and imaging data of 12 patients with EP who were admitted to Xuanwu Hospital of Capital Medical University from January 2017 to June 2020 were collected. There were 10 males and 2 females, aged from 25 to 71 years, with a median age of 42 years. All patients received CT examination. Step-up treatment or one-step surgical treatment was performed on patients according to their conditions. Observation indicators: (1) CT features; (2) bacteriological characteristics; (3) treatment and follow-up. Follow-up using outpatient examination was conducted at postoperative 1, 3, 6 months to detect survival of patients up to January 2021. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers. Results:(1) CT features: 1 of the 12 patients underwent abdominal+pelvic CT plain scan, and 11 cases underwent abdominal+pelvic CT plain scan and enhanced scan of arterial and portal venous phase. CT examination of 12 patients showed diffuse enlargement of the pancreas, unclear borders and a large amount of exudation around the pancreas. Pancreatic necrotic tissues accounted for >30% of the total pancreatic volume; the Balthazar CT score was 10 (range, 8-10). Of the 12 patients, 5 cases showed that the exudation or necrosis involved bilateral prerenal fascia, 7 cases only involved the left prerenal fascia; the necrotic infection area of 11 patients formed obvious wraps. The distribution of pancreatic, peripancreatic infection and gas in 12 patients: 6 cases had pancreatic, peripancreatic infection and gas located in Ⅰ+Ⅱa area, 3 cases located in Ⅰ+Ⅱa+Ⅲ area, 2 cases located in Ⅰ+Ⅲ area, and 1 case located in Ⅰ area. There was gas in the pancreatic parenchyma in 12 patients, with fluid in the abdominal cavity and pelvic cavity. (2) Bacteriological characteristics: the culture results of peripancreatic necrotic issues in 12 patients were all positive for the pathogenic specimens, and 27 strains were cultured. Klebsiella pneumoniae was the most common in the culture of necrosis from 12 patients, followed by Escherichia coli and Enterococcus bacteria. Fungus was found in the culture of necrosis from 1 patient. Of the 12 patients, 5 had negative blood cultures and 7 had positive blood cultures. A total of 14 strains were cultured, with Klebsiella pneumoniae being the most common; fungus was found in the blood culture from 4 patients. (3) Treatment and follow-up: 1 patient underwent percutaneous catheter drainage, 7 underwent step-up surgical treatment, 4 underwent one-step surgical treatment; 11 patients undergoing surgical treatment received laparoscopic-assisted removal of pancreatic necrotic tissue, including 1 case with exploratory laparotomy due to abdominal hemorrhage. Of the 11 patients undergoing surgical treatment, 7 cases received the left retroperitoneal approach surgery (including 1 case combined with the upper abdominal median approach), 2 cases received the upper abdominal median transomental sac approach surgery, 1 case received the right retroperitoneal approach surgery, and 1 case received the left rectus abdominis approach surgery. The number of operations of all the 11 patients were (3.1±0.9)times, the number of step-up treatments was (3.6±0.8)times, and the number of one-step surgery was (2.3±0.5)times. Nine of 12 patients had organ dysfunction that lasted for more than 48 hours during the treatment, which received surgical treatment after organ support and anti-infection therapy. All the 12 patients were followed up for 6 months after operation, of which 9 cases were cured after treatment and 3 cases died including 1 case dying of bleeding and 2 cases dying of septic shock combined with multiple organ failure.Conclusions:Emphysema pancreatitis is complicated by pancreatic necrosis, which is characterized by pancreatic and peripancreatic gas accumulation on CT. Most patients with EP have organ failure. Surgery is an important treatment for EP.