Effect of Wnt/β-catenin signaling pathway in neural differentiation of human bone marrow mesenchymal stem cells / 中国修复重建外科杂志

OBJECTIVE@#To explore the effect of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and the combination of bFGF and EGF in the neural differentiation of human bone marrow mesenchymal stem cells (hBMSCs), and the role of Wnt/β-catenin signaling pathway in this process.@*METHODS@#The identified 4th-generation hBMSCs were divided into five groups according to different induction conditions, namely control group (group A), EGF induction group (group B), bFGF induction group (group C), EGF and bFGF combined induction group (group D), and EGF, bFGF, and Dickkopf-related protein 1 (DKK-1) combined induction group (group E). After 7 days of continuous induction, the cell morphology was observed by inverted fluorescence phase contrast microscopy, levels of genes that were related to neural cells [Nestin, neuron-specific enolase (NSE), microtubule-associated protein 2 (MAP-2), and glial fibrillary acidic protein (GFAP)] and key components of the Wnt/β-catenin signaling pathway (β-catenin and Cyclin D1) were detected by RT-PCR, and the levels of proteins that were related to neural cells (Nestin and GFAP) as well as genes that were involved in Wnt/β-catenin signaling pathway [β-catenin, phosphorylation β-catenin (P-β-catenin), Cytoplasmic β-catenin, and Nuclear β-catenin] were explored by cellular immunofluorescence staining and Western blot.@*RESULTS@#When compared to groups A and B, the typical neuro-like cell changes were observed in groups C-E, and most obviously in group D. RT-PCR showed that the relative expressions of Nestin, NSE, and MAP-2 genes in groups C-E, the relative expressions of GFAP gene in groups D and E, the relative expression of NSE gene in group B, the relative expressions of β-catenin gene in groups C and D, and the relative expressions of Cyclin D1 gene in groups B-D significantly increased when compared with group A ( P<0.05). Compared with group E, the relative expressions of Nestin, NSE, MAP-2, GFAP, β-catenin, and CyclinD1 genes significantly increased in group D ( P<0.05); compared with group C, the relative expression of Nestin gene in group D significantly decreased ( P<0.05), while NSE, MAP-2, and GFAP genes significantly increased ( P<0.05). The cellular immunofluorescence staining showed that the ratio of NSE- and GFAP-positive cells significantly increased in groups C-E than in group A, in group D than in groups C and E ( P<0.05). Western blot assay showed that the relative expression of NSE protein was significantly higher in groups C and D than in group A and in group D than in groups C and E ( P<0.05). In addition, the relative expression of GFAP protein was significantly higher in groups C-E than in group A and in group D than in group E ( P<0.05). Besides, the relative expressions of β-catenin, Cytoplasmic β-catenin, Nuclear β-catenin, and the ratio of Nuclear β-catenin to Cytoplasmic β-catenin were significantly higher in groups C and D than in group A and in group D than in group E ( P<0.05), whereas the relative expression of P-β-catenin protein was significantly lower in groups C and D than in group A and in group D than in group E ( P<0.05).@*CONCLUSION@#Different from EGF, bFGF can induce neural differentiation of hBMSCs. In addition, EGF can enhance the hBMSCs neural differentiation of bFGF, while the Wnt/β-catenin signaling pathway may play a positive regulatory role in these processes.

Physical activity of Chinese children and adolescents:age, gender and regional characteristics / 中国学校卫生

Objective@#To explore age, gender, and regional differences in physical activity among children and adolescents in China, and to provide a scientific reference for enhancing physical activity promotion.@*Methods@#A total of 4 269 children and adolescents aged 7 to 18 years were selected from six administrative regions of China (East China, Northwest China, North China, Central China, Southwest China and South China) using a stratified random cluster sampling method from September to December 2018. A questionnaire was administered to evaluate the physical activity level of Chinese children and adolescents aged 7 to 18.@*Results@#The overall detection rate of MVPA insufficiency in children and adolescents in China was 53.8%, of which the detection rate of MVPA insufficiency was 50.8% among boys and 57.1% among girls. Gender differences were statistically significant ( χ 2= 17.10 , P <0.05). Among the different age groups, the lowest detection rate of MVPA among 10-12 year olds was 43.6%, whereas the highest rate among 16-18 year olds was 63.0%, with significant differences between gender ( χ 2=4.33, 30.79, P <0.05). The P 50 values of total physical activity(TPA), light intensity physical activity(LPA), moderate intensity physical activity(MPA), vigorous intensity physical activity(VPA), moderate to vigorous physical activity(MVPA) were 92.9,24.3,41.4,7.1 and 55.7 min/d , respectively. The P 50 values of physical exercise, housework activities, entertainment activities and transportation activities were 34.3 , 2.1, 2.3 and 30.0 min/d, respectively, and the difference in age groups was statistically significant( H =95.03, 74.99, 300.26 , 64.16, P <0.05). There was a statistically significant difference in the detection rate of insufficient MVPA among children and adolescents in different regions ( χ 2=83.91, P <0.05). The lowest rate was 44.0% in North China, and the highest was 65.9% in East China.@*Conclusion@#The detection rate of MVPA insufficiency among Chinese children and adolescents firstly decreased and then increased with age. Boys participated in higher levels of physical activity than girls.

Application of acellular bovine pericardium patch in implant based immediate breast reconstruction / 中华外科杂志

Objective: To examine the clinical effect of acellular bovine pericardium patch in implant based immediate breast reconstruction. Methods: The clinicopathological information of 141 breast cancer patients, who admitted to Department of Breast Reconstruction and Oncoplastic Surgery, Tianjin Medical University Cancer Hospital, underwent immediate mammoplasty with implants combined with acellular bovine pericardium patches were analyzed from June 2016 to October 2019. All patients were female, with the age of (38.8±8.5) years (range: 13 to 60 years). The body mass index was (21.9±2.5) kg/m2 (range: 16.0 to 32.3 kg/m2). There were 39 cases of duct carcinoma in situ, 46 cases of stage Ⅰ, 40 cases of stage Ⅱ and 16 cases of stage Ⅲ. All patients received nipple-areola-sparing mastectomy or skin-sparing mastectomy with sentinel lymph node biopsy or axillary lymph node dissection, and prosthesis implantation with sub-pectoralis combined with breast patch. The correlation of clinicopathological characters and complications was assessed by t test, χ2 test, Fisher's exact probability method and Logistic regression. Pre-and post-operative aesthetic, quality of life scores were recorded. Results: The operation time (M(IQR)) was 3.6(1.5) hours (range: 3.0 to 6.5 hours). The early postoperative complication rate was 22.0% (31/141), prosthesis removal was the main postoperative complication, accounting for 64.5% (20/31) of the total complications, of which 15 cases occurred in the first 30 patients. The follow-up time was 28(8) months (range: 20 to 53 months), The most frequent long-term complications were capsular contracture and implant displacement, with the incidence of 11.2% (14/125) and 10.4% (13/125), respectively. Multivariate analysis showed that prosthesis volume ≥300 ml (OR=8.173, 95%CI: 1.302 to 51.315, P=0.021) and peri-areolar incision (OR=7.809, 95%CI: 2.162 to 28.211, P<0.01) were independent relative factors for the occurrence of short-term postoperative local complications. After 2 years of operation, the score of breast appearance satisfaction was 71.7±15.5, postoperative effect satisfaction was 90.4±9.5, psychological satisfaction was 90.7±17.1, sexual satisfaction was 70.1±25.1. The immediate postoperative satisfaction rate at discharge was 95.4% (134/141), and 17.6% (22/125) of patients had the intention to received revision surgery. Conclusions: Prosthesis volume ≥300 ml and peri-areolar incision were independent realtive factors for short-term local complications after bovine pericardium patch combined with prosthesis implantation in the immediate breast reconstruction. After completing the learning curve, the postoperative complications of the procedure could be decreased.

Coactivator p300-induced H3K27 acetylation mediates lipopolysaccharide-induced inflammatory mediator synthesis / 南方医科大学学报

OBJECTIVE@#To investigate the role of acetylated modification induced by coactivator p300 in lipopolysaccharide (LPS)- induced inflammatory mediator synthesis and its molecular mechanism.@*METHODS@#Agilent SurePrint G3 Mouse Gene Expression V2 microarray chip and Western blotting were used to screen the molecules whose expression levels in mouse macrophages (RAW246.7) were correlated with the stimulation intensity of LPS. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (chip-qPCR) were used to verify the binding of the molecules to the promoters of IL-6 and TNF-α genes. The effects of transfection of RAW246.7 cells with overexpression or interfering plasmids on IL-6 and TNF-α synthesis were evaluated with ELISA, and the binding level of the target molecules and acetylation level of H3K27 in the promoter region of IL-6 and TNF-α genes were analyzed by chromatin immunoprecipitation sequencing technique (chip-seq).@*RESULTS@#Gene microarray chip data and Western blotting both confirmed a strong correlation of p300 expression with the stimulation intensity of LPS. Immunocoprecipitation confirmed the binding between p300 and c-myb. The results of EMSA demonstrated that c-myb (P < 0.05), but not p300, could directly bind to the promoter region of IL-6 and TNF-α genes; p300 could bind to the promoters only in the presence of c-myb (P < 0.05). The expressions of p65, p300 and c-myb did not show interactions. Both p300 overexpression and LPS stimulation could increase the level of promoter-binding p300 and H3K27 acetylation level, thus promoting p65 binding and inflammatory gene transcription; such effects were obviously suppressed by interference of c-myb expression (P < 0.05). Interference of p65 resulted in inhibition of p65 binding to the promoters and gene transcription (P < 0.05) without affecting p300 binding or H3K27 acetylation level.@*CONCLUSION@#LPS can stimulate the synthesis of p300, whose binding to the promoter region of inflammatory genes via c-myb facilitates the cohesion of p65 by inducing H3K27 acetylation, thus promoting the expression of the inflammatory genes.

Molecular Mechanism of Jingfang Mixture Against H1N1 Influenza Based on Network Pharmacology and Experimental Verification / 中国实验方剂学杂志

ObjectiveTo predict the potential targets and mechanism of Jingfang mixture in the treatment of H1N1 influenza and provide references for clinical application of Jingfang mixture. MethodThe active components and targets of Jingfang mixture against H1N1 influenza were screened out by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP），SwissTargetPrediction， and TargetNet. The targets of H1N1 influenza were obtained from GeneCards，Online Mendelian Inheritance in Man （OMIM）， and DisGeNET and standardized by UniProt KB. The intersection targets were obtained by Venny 2.1.0. The "drug-component-target" network was constructed with Cytoscape 3.2.1 and analyzed for the topological attributes. The intersection targets were uploaded to STRING 11.5 to obtain the protein-protein interaction （PPI） network. Gene Ontology （GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） analysis were carried out by Metascape. Finally，the top active components ranked by degree were docked to the core targets by Autodock vina and visually analyzed by PyMOL. Balb/c female rats were used for experimental verification. Hematoxylin-eosin（HE） staining was used to observe the pathological changes in lung tissues. Enzyme-linked immunosorbent assay（ELISA）was used to detect the levels of tumor necrosis factor-α（TNF-α），interleukin-10（IL-10）， and interleukin-17（IL-17）. Real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR） and Western blot were used to detect the mRNA and protein expression levels in lung tissues. ResultThere were 144 active components in Jingfang mixture. A total of 421 target genes of Jingfang mixture and 2 956 targets of H1N1 influenza were identified，including 199 common targets. Topological analysis showed that the core components of Jingfang mixture against H1N1 influenza included quercetin，luteolin， and kaempferol，and the core targets included prostaglandin-endoperoxide synthase 2（PTGS2），estrogen receptor alpha（ESR1），inducible nitric oxide synthase 2（iNOS2），peroxisome proliferator-activated receptorγ（PPARγ），and cyclooxygenase-1（PTGS1）. GO enrichment yielded 697 items in biological process （BP） （P<0.01）， 59 items in molecular function （MF）（P<0.01）， and 21 items in cellular component （CC） （P<0.01）. A total of 132 signaling pathways （P<0.01） were obtained by KEGG enrichment analysis， including phosphatidylinositol 3-kinases（PI3K）/protein kinase B（Akt） signaling pathway and mitogen-activated protein kinase（MAPK） signaling pathway，most of which were related to the regulation of immune inflammation. Molecular docking showed that the binding energy of the active components of Jingfang mixture to the core targets was less than -5.0 kcal·mol-1，indicating good binding activity. HE staining showed that the lung tissues were significantly improved after drug intervention，and Real-time PCR and Western blot showed that Jingfang mixture could reduce the mRNA and protein expression of PI3K and Akt in lung tissues. ConclusionJingfang mixture can play an anti-viral effect against the influenza A virus through multiple components，multiple targets， and multiple pathways. The active components quercetin，luteolin， and kaempferol may control the inflammation and regulate immunity on the PI3K/Akt，MAPK， and other signaling pathways by acting on targets such as PTGS2，ESR1，iNOS2，PPARγ， and PTGS1.

Study on quality standard of Jingtian Zhitong cream / 药学实践杂志

Objective To revise the determination method in the quality standard of Jingtian Zhitong cream. Methods The total saponins of angelica sinensis, Ligusticum wallichii, Rhizoma corydalis, and Panax notoginseng saponins were qualitatively identified by thin-layer chromatography (TLC). The contents of notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1 in the preparation were determined by high performance liquid chromatography (HPLC). Results TLC showed strong specificity and good resolution. The concentration of notoginsenoside R1 showed a good linear relationship in the range of 0.1604 and 2.0050μg (r=0. 999). The concentration of ginsenoside Rg1 showed a good linear relationship in the range of 0.8003 and 10.0035μg (r=1.000). The concentration of ginsenoside Rb1 showed good linearity in the range of 0.6182 and 7.7275μg (r=1.000). The sample recovery rates were 101.43%, 98.75% and 100.95%, respectively. The relative standard deviation (RSD) were 2.56%, 2.71% and 2.75%, respectively. Conclusion The developed method is accurate and reliable with high sensitivity, which can be used for the quality control of Jingtian Zhitong cream.

Research advances in animal models of hepatitis B virus infection / 临床肝胆病杂志

For the ideal preclinical animal model of hepatitis B virus (HBV), its hepatocytes should allow HBV entry and cccDNA generation and have both innate and adaptive immune systems. However, HBV only naturally infects humans and chimpanzees due to highly restricted species specificity, and no effective model has been established so far to truly reflect the immune mechanism and pathogenesis of HBV infection. This article reviews five commonly used mouse models, i.e., HBV transgenic model, HBV plasmid DNA hydrodynamic injection model, AAV-HBV transfection model, cccDNA surrogate model, and human-mouse chimeric liver model, and looks forward to the new models that will appear in the future, such as hNTCP transgenic cynomolgus monkey, rhesus monkey, or pig models, so as to provide a reference for researchers to select these models, accelerate the process of drug screening, validate new therapies, and better solve the problems of HBV biological pathogenesis.

Characteristics of polysomnography in depression patients complicating moderate-to-severe obstructive sleep apnea hypopnea syndrome / 四川精神卫生

ObjectiveTo investigate the characteristics of polysomnography （PSG） in depression patients complicating moderate-to-severe obstructive sleep apnea hypopnea syndrome （OSAHS）. MethodsA retrospective analysis was conducted on the outpatients， inpatients and physical examination population who completed overnight PSG monitoring in the sleep medicine center of Suzhou Guangji Hospital from December 2017 to October 2019. Four groups of subjects were finally enrolled， including depression patients with moderate-to-severe OSAHS （n=31）， depression patients without OSAHS （n=79）， moderate-to-severe OSAHS patients （n=96） and normal control group （n=32）. The sleep process related indicators （total sleep time， sleep latency， number of awakenings）， sleep structure related indicators （N1， N2， N3， percentage of REM sleep， REM latency， REM sleep duration）， sleep-related respiratory variables （oxygen reduction index） and other polysomnographic parameters of the four groups were compared. ResultsIn terms of sleep process， the total sleep time， sleep latency and number of awakenings yielded significant differences among the four groups （F=2.874， 3.959， 12.291， P<0.05 or 0.01）. In terms of sleep structure， the percentage of total sleep time in N2 and N3 stages demonstrated significant differences among the four groups （F=13.885， 48.013， P<0.01）. The REM latency， REM sleep duration and percentage of REM sleep manifested significant differences among the four groups （F=41.492， 11.827， 10.552， P<0.01）. In terms of sleep-related respiratory variables， the oxygen reduction index exhibited significant differences among the four groups （F=170.585， P<0.05）. ConclusionDepression patients complicating moderate-to-severe OSAHS suffer from severe sleep process and structural disturbances， accompanied by quite frequent and severe sleep-related respiratory events.

Effect of Petroleum Ether Extract of Rhizoma Amorphophalli on Biological Characteristics of K562 Cells and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the role of petroleum ether extract of Rhizoma Amorphophalli (SLG) in inhibiting proliferation and promoting apoptosis and differentiation of leukemia K562 cells.@*METHODS@#K562 cells were processed by SLG and PD98059 which was the ERK signaling pathway blocker. Then cell vitality was tested by MTT. Cell apoptosis rate and positive percentage of antigen expression related with differentiation were detected by flow cytometry. The protein expression levels of ERK1/2 and pERK1/2 were detected by Western blot.@*RESULTS@#The proliferation activity of K562 was reduced by 50, 100, 200 mg/L SLG in a concentration dependent manner (r=0.9997). The apoptosis rate and positive expression rate of CD11b, CD14 and CD42b which were related with differentiation were raised by SLG, as well as the expression of pERK1/2, while PD98059 could reverse the promoting effect of SLG on apoptosis and differentiation partially.@*CONCLUSION@#SLG can inhibit the proliferation and promote apoptosis and differentiation of K562 cells through ERK signaling pathway.

Natural Orifice Specimen Extraction (NOSE) with SingleStapling Anastomosis for Left Colon Cancer

One of the major criticisms of laparoscopic colectomy is the requirement of an additional mini laparotomy to remove the resected specimen. This may be associated with postoperative pain and wound complications. Therefore, the use of a natural orifice as a delivery route for the specimen extraction without a laparotomy incision can subsequently reduce the risk of wound complications. In this video, we demonstrate the natural orifice specimen extraction procedure with a side-to-end single-stapling colorectal anastomosis.

HBV induces liver fibrosis through regulating TGF-β1/Nur77 pathway / 医学研究生学报

ObjectiveTo investigate the role of TGF-β1/Nur 77 in HBV-induced liver fibrosis.MethodsHE, Masson's and immunohistochemistry (IHC) staining were used to detect Nur 77 expression in liver tissue. NTCP-Huh7.5.1 was constructed by transfecting pCMV6-NTCP into Huh 7.5.1 cells to explore effects of HBV replication and TGF-β1 expression. LX-2 cells was incubated with TGF-β1 in the presence of Nur 77 gRNA. QPCR was used to observe the effect of TGF-β1/Nur 77 on LX-2 activation and liver fibrosis. Our study engaged a co-culture system in the presence of Nur 77 gRNA to observe the relationship between HBV replication andα-SMA, TIMP-1 and CoL1A1 expression.ResultsIHC showed a higher Nur 77 level in severe fibrosis liver (≥S2) than in mild fibrosis liver (≤S1). Western blot showed that pCMV6-NTCP could express in Huh 7.5.1 cells. At 3 days after infection, HBV DNA level in supernatant of NTCP-Huh 7.5.1 cells was 5.39±0.96×103 copies/ml (P<0.01), HBV infection enhanced TGF-β1 mRNA expression in NTCP-Huh 7.5.1 cells (1.94 vs 1.00，P<0.01). At 72 h after TGF-β1 incubation, mRNA expression of Nur 77, α-SMA, TIMP-1 and CoL1A1 expression was up-regulated significantly compared to control group (P<0.05), which can be inhibited by Nur 77 gRNA (P<0.05). Co-cultured with HBV infected NTCP-Huh 7.5.1 cells for 72 h, mRNA expression of Nur 77, α-SMA, TIMP-1 and CoL1A1 expression in LX-2 cells was up-regulated significantly compared to control group (P<0.05), which can be inhibited by Nur 77 gRNA (P <0.05).ConclusionHBV contributes to liver fibrosis through up-regulating TGF-β1/Nur 77 signaling pathway. Our study provides a new target for the treatment of HBV induced liver fibrosis.

Natural Orifice Specimen Extraction (NOSE) with SingleStapling Anastomosis for Left Colon Cancer

One of the major criticisms of laparoscopic colectomy is the requirement of an additional mini laparotomy to remove the resected specimen. This may be associated with postoperative pain and wound complications. Therefore, the use of a natural orifice as a delivery route for the specimen extraction without a laparotomy incision can subsequently reduce the risk of wound complications. In this video, we demonstrate the natural orifice specimen extraction procedure with a side-to-end single-stapling colorectal anastomosis.

Prevalence of chronic diseases and associate factors on daily activities in male oldest-olds / 中华流行病学杂志

Objective@#To investigate the prevalence of chronic diseases in aged ≥80 oldest-olds and related factors influencing their daily activities.@*Methods@#This survey was conducted in the retired cadres in Beijing from 2012 to 2014. A unified questionnaire was used to investigate the general characteristics of the oldest-olds and the activities of daily living (ADL). Information on chronic diseases was extracted from related medical records.@*Results@#A total of 4 472 male oldest- olds, with an average age as (87.1±3.9) years (80-102 years), were included. Nearly half of the elderly people were suffering from 5 or more kinds of chronic diseases, with 43.9% of them having disability on basic daily activities (BADL) with 13.4% of those classified as moderate or severe cases. 38.8% of them had instrumental activities of daily living (IADL) disability, with 28.7% of them were moderate or severe cases. The ADL disability showed an increasing trend along with the increase number of chronic diseases. The proportion of BADL disability increased from 40.5% to 50.6%. Compared with the ones having fewer chronic diseases (≤2 kinds), those with more (≥7 kinds) had an increase of 50.5% risk on BADL disability and 199.4% on IADL disability.@*Conclusion@#We noticed that the male oldest-olds suffered from multiple chronic diseases. The impairment of ADL was higher than the younger elderly. Comorbidity showed heavier impact on ADL, especially on the instrumental activities of daily living.

Treament of CRRT combined with low-flow ECMO for canines with ARDS / 医学研究生学报

Objective Single mechanical ventilation for curing severe acute respiratory distress syndrome (ARDS) is sometimes not effective in improving oxygenation and removing excess CO2 in the blood. The purpose of this study is to observe the effect of continuous renal replacement therapy（CRRT）combined with low-flow extracorporeal membrane oxygenation （ECMO）of V-V mode on clearance of inflammatory mediators，improving oxygenation and reducing PaCO2 in canines with ARDS and hypercapnia. Methods 30 healthy adult canines were divided into 3 groups (n=10 each) using a random number table method sham group, ECMO (EC) group and CRRT+ECMO（CR+EC）group. A canine model of ARDS with hypercapnia were prepared by injected intravenously oleic acid. Sham group was only treated with invasive mechanical ventilation. EC group was treated with invasive mechanical ventilation and ECMO. CR+EC group was treated with CRRT combined with low-flow ECMO of V-V mode besides invasive mechanical ventilation. The heart rate(HR)，mean artery pressure (MAP)，cardiac output(CO) and concentration of serum TNF-α and IL-6 were examined at the beginning of modeling(T0)，after 1，3，6 and 9 (T1, T3, T6, T9) hours of mechanical treatment respectively. Oxygenation index（OI），PaCO2, temperature（T）were examined at the beginning of modeling，after 3, 6, 9 hours of mechanical treatment，and at the time point of 3 hours(T12) after ECMO treatment (EC group) and CRRT+ECMO treatment(CR+EC group)，respectively. The differences were compared between groups involved in above-mentioned indexes at different time points. Results Compared with the sham group, the HR decreased at T6 and T9 in the EC group and CR+EC group (P<0.05), and the concentration of IL-6 and TNF-α decreased at T3, T6 and T9 in the CR+EC group (P<0.05). Compared with T0, HR decreased at T6 and T9 in the EC group (P<0.05). Compared with T0, the concentration of IL-6 and TNF-α decreased at T3, T6 and T9 in the CR+EC group (P<0.05). Compared with the EC group, the HR decreased at T6 and T9 in the CR+EC group (P<0.05). Compared with the EC group, the concentration of IL-6 and TNF-α decreased at T3, T6 and T9 in the CR+EC group (P<0.05). Compared with the sham group, the OI decreased at T9 and T12 in the CR+EC group (P<0.05). Compared with the sham group, PaCO2 decreased at T3, T6 and T9 in the EC group and CR+EC group (P<0.05). Compared with the EC group, the OI decreased at T9 and T12 in the CR+EC group (P<0.05). Compared with T0, the OI decreased at T6 and T12 in the CR+EC group (P<0.05). Conclusion The CRRT combined with low-flow ECMO of V-V mode has positive effects on clearance of inflammatory mediators，reducing hypercapnia and improving oxygenation in the canines with ARDS and hypercapnia.

Clinical association between pre-treatment levels of plasma fibrinogen and bone metastatic burden in newly diagnosed prostate cancer patients / 中华医学杂志(英文版)

Background@#Due to the different treatments for low-volume metastatic prostate cancer (PCa) as well as high-volume ones, evaluation of bone metastatic status is clinically significant. In this study, we evaluated the correlation between pre-treatment plasma fibrinogen and the burden of bone metastasis in newly diagnosed PCa patients.@*Methods@#A single-center retrospective analysis, focusing on prostate biopsies of newly diagnosed PCa patients, was performed. A total of 261 patients were enrolled in this study in a 4-year period. All subjects were submitted to single-photon emission computerized tomography-computed tomography to confirm the status of bone metastasis and, if present, the number of metastatic lesions would then be calculated. Clinical information such as age, prostate-specific antigen (PSA), fibrinogen, clinical T stage, and Gleason score were collected. Patients were divided into three groups: (i) a non-metastatic group, (ii) a high volume disease (HVD) group (>3 metastases with at least one lesion outside the spine), and (iii) a low volume disease (LVD) group (metastatic patients excluding HVD ones). The main statistical methods included non-parametric Mann-Whitney test, Spearman correlation, receiver operating characteristic (ROC) curves, and logistic regression.@*Results@#Fibrinogen positively correlated with Gleason score (r = 0.180, P = 0.003), PSA levels (r = 0.216, P < 0.001), and number of metastatic lesions (r = 0.296, P < 0.001). Compared with the non-metastatic and LVD groups, the HVD group showed the highest PSA (104.98 ng/mL, median) and fibrinogen levels (3.39 g/L, median), as well as the largest proportion of Gleason score >7 (86.8%). Both univariate (odds ratio [OR] = 2.16, 95% confidential interval [CI]: 1.536-3.038, P < 0.001) and multivariate (OR = 1.726, 95% CI: 1.206-2.472, P = 0.003) logistic regressions showed that fibrinogen was independently associated with HVD. The ROC curve suggested that fibrinogen acts as a predictor of HVD patients, yielding a cut-off of 3.08 g/L, with a sensitivity of 0.684 and a specificity of 0.760 (area under the curve = 0.739, 95% CI: 0.644-0.833, P < 0.001).@*Conclusions@#Pre-treatment plasma fibrinogen is positively associated with bone metastatic burden in PCa patients. Our results indicate that fibrinogen might be a potential predictor of HVD.

The efficacy analysis of allogeneic hematopoietic stem cell transplantation in 48 leukemia patients with central nervous system leukemia / 中华血液学杂志

Objective: To investigate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of leukemia patients also suffering from central nervous system leukemia (CNSL) . Methods: A total of 48 leukemia patients with central nervous system leukemia admitted to our hospital from May 2012 to December 2017 were retrospectively analyzed. Results: ① Including 22 cases of acute lymphocytic leukemia (ALL) , 21 cases of acute myeloid leukemia (AML) , and 5 cases of chronic myelogenous leukemia (CML) . Before transplantation, 19 patients achieved complete remission (CR) , and the rest 29 ones without remission. ②The conditioning regimen used TBI as the main protocol, and 6 patients were combined with whole brain and total spinal cord radiotherapy, 2 with Cyber knife treatment, and children with modified IDA combined with BUCY. ③All 48 patients were successfully transplanted, the median time for leukocyte engraftment was 14 (10-23) days, the median time for platelet transplant 16 (6-78) days. ④Bone marrow was evaluated 28 days after transplantation, all 48 patients reached CR, and DNA testing confirmed that they were all full donor chimerism. ⑤The median follow-up was 14 (2-69) months. Of them, 28 cases survived, 10 relapsed and the rest 3 had recurrence of CNSL after transplantation. One year after allo-HSCT, the overall survival (OS) of CR and non-CR groups were (77.3±10.0) % and (57.6±9.3) % (P=0.409) , respectively, the disease-free survival rates (DFS) were (71.2±11.0) % and (53.9±9.5) % (P=0.386) , respectively. The 1-year OS rates of ALL and AML groups after transplantation were (54.2±10.7) %, (80.1±8.9) %, respectively (P=0.200) , and DFS rates were (49.2±10.8) %, (75.0±9.7) % (P=0.190) , respectively. Conclusion: Allo-HSCT was safe and effective for leukemia patients with CNSL.

Clinical association between pre-treatment levels of plasma fibrinogen and bone metastatic burden in newly diagnosed prostate cancer patients / 中华医学杂志(英文版)

BACKGROUND@#Due to the different treatments for low-volume metastatic prostate cancer (PCa) as well as high-volume ones, evaluation of bone metastatic status is clinically significant. In this study, we evaluated the correlation between pre-treatment plasma fibrinogen and the burden of bone metastasis in newly diagnosed PCa patients.@*METHODS@#A single-center retrospective analysis, focusing on prostate biopsies of newly diagnosed PCa patients, was performed. A total of 261 patients were enrolled in this study in a 4-year period. All subjects were submitted to single-photon emission computerized tomography-computed tomography to confirm the status of bone metastasis and, if present, the number of metastatic lesions would then be calculated. Clinical information such as age, prostate-specific antigen (PSA), fibrinogen, clinical T stage, and Gleason score were collected. Patients were divided into three groups: (i) a non-metastatic group, (ii) a high volume disease (HVD) group (>3 metastases with at least one lesion outside the spine), and (iii) a low volume disease (LVD) group (metastatic patients excluding HVD ones). The main statistical methods included non-parametric Mann-Whitney test, Spearman correlation, receiver operating characteristic (ROC) curves, and logistic regression.@*RESULTS@#Fibrinogen positively correlated with Gleason score (r = 0.180, P = 0.003), PSA levels (r = 0.216, P 7 (86.8%). Both univariate (odds ratio [OR] = 2.16, 95% confidential interval [CI]: 1.536-3.038, P < 0.001) and multivariate (OR = 1.726, 95% CI: 1.206-2.472, P = 0.003) logistic regressions showed that fibrinogen was independently associated with HVD. The ROC curve suggested that fibrinogen acts as a predictor of HVD patients, yielding a cut-off of 3.08 g/L, with a sensitivity of 0.684 and a specificity of 0.760 (area under the curve = 0.739, 95% CI: 0.644-0.833, P < 0.001).@*CONCLUSIONS@#Pre-treatment plasma fibrinogen is positively associated with bone metastatic burden in PCa patients. Our results indicate that fibrinogen might be a potential predictor of HVD.

Correction to: Efficient derivation of extended pluripotent stem cells from NOD-scid Il2rg mice

In the original publication Fig. 1D and supplementary material is incorrect. The correct figure and supplementary material is provided in this correction.

Efficient derivation of extended pluripotent stem cells from NOD-scid Il2rg mice

Recently we have established a new culture condition enabling the derivation of extended pluripotent stem (EPS) cells, which, compared to conventional pluripotent stem cells, possess superior developmental potential and germline competence. However, it remains unclear whether this condition permits derivation of EPS cells from mouse strains that are refractory or non-permissive to pluripotent cell establishment. Here, we show that EPS cells can be robustly generated from non-permissive NOD-scid Il2rg mice through de novo derivation from blastocysts. Furthermore, these cells can also be efficiently generated by chemical reprogramming from embryonic NOD-scid Il2rg fibroblasts. NOD-scid Il2rg EPS cells can be expanded for more than 20 passages with genomic stability and can be genetically modified through gene targeting. Notably, these cells contribute to both embryonic and extraembryonic lineages in vivo. More importantly, they can produce chimeras and integrate into the E13.5 genital ridge. Our study demonstrates the feasibility of generating EPS cells from refractory mouse strains, which could potentially be a general strategy for deriving mouse pluripotent cells. The generation of NOD-scid Il2rg EPS cell lines permits sophisticated genetic modification in NOD-scid Il2rg mice, which may greatly advance the optimization of humanized mouse models for biomedical applications.

Panaxdiol Saponins Component Promotes Hematopoiesis and Modulates T Lymphocyte Dysregulation in Aplastic Anemia Model Mice / 中国结合医学杂志

OBJECTIVE@#To investigate the potential efficacy of panaxadiol saponins component (PDS-C) in the treatment of aplastic anemia (AA) model mice.@*METHODS@#Totally 70 mice were divided into 7 groups as follows: normal, model, low-, medium-, high-dose PDS-C (20, 40, 80 mg/kg, namely L-, M-, H-PDS-C), cyclosporine (40 mg/kg), and andriol (25 mg/kg) groups, respectively. An immune-mediated AA mouse model was established in BALB/c mice by exposing to 5.0 Gy total body irradiation at 1.0 Gy/min, and injecting with lymphocytes from DBA mice. On day 4 after establishment of AA model, all drugs were intragastrically administered daily for 15 days, respectively, while the mice in the normal and model groups were administered with saline solution. After treatment, the peripheral blood counts, bone marrow pathological examination, colony forming assay of bone marrow culture, T lymphocyte subpopulation analysis, as well as T-bet, GATA-3 and FoxP3 proteins were detected by flow cytometry and Western blot.@*RESULTS@#The peripheral blood of white blood cell (WBC), platelet, neutrophil counts and hemoglobin (Hb) concentration were significantly decreased in the model group compared with the normal group (all P<0.01). In response to 3 dose PDS-C treatment, the WBC, platelet, neutrophil counts were significantly increased at a dose-dependent manner compared with the model group (all P<0.01). The myelosuppression status of AA was significantly reduced in M-, H-PDS-C groups, and hematopoietic cell quantity of bone marrow was more abundant than the model group. The colony numbers of myeloid, erythroid and megakaryocytic progenitor cells in the model group were less than those of the normal mice in bone marrow culture, while, PDS-C therapy enhanced proliferation of hematopoietic progenitor cells by significantly increasing colony numbers (all P<0.01). Furthermore, PDS-C therapy increased peripheral blood CD3 and CD3CD4 cells and reduced CD3CD8 cells (P<0.05 or P<0.01). Meanwhile, PDS-C treatment at medium- and high doses groups also increased CD4CD25FoxP3 cells, downregulated T-bet protein expression, and upregulated GATA-3 and FoxP3 protein expressions in spleen cells (P<0.05).@*CONCLUSION@#PDS-C possesses dual activities, promoting proliferation hematopoietic progenitor cells and modulating T lymphocyte immune functions in the treatment of AA model mice.