This paper reported a case of severe COVID-19 in the recovery stage with acute lymphoblastic leukemia treated by integrated traditional Chinese and western medicine， with the intention of shedding light on the clinical diagnosis and treatment of similar conditions. The patient， who had acute lymphoblastic leukemia， developed COVID-19 infection during the bone marrow suppression period after chemotherapy. Treatment with western medicine was mainly anti-infection， symptomatic management， and supportive care. During the recovery stage， considering the patient's chemotherapy history and disease progression， the overall syndrome was identified as deficiency of both qi and yin and binding of phlegm and blood. Based on the “state-target” combined treatment strategy， herbal prescriptions were selected and modified to address the “deficiency state”， “disease target”， and “symptom target”. In addition to western medicine， the patient was administered with Shengmai Powder （生脉散） and Compound Zhebei Granules （复方浙贝颗粒） in its modifications to boost qi， nourish yin， and reinforce healthy qi， nourish and cool the blood， ultimately achieving satisfactory therapeutic effects.

Improving the reproducibility of biomedical research results is a major challenge.Transparent and accurate reporting of the research process enables readers to evaluate the reliability of the research results and further explore the experiment by repeating it or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), provide a checklist applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as the reliability, repeatability, and clinical translatability of animal experimental results. The use of ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and integrity of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is a Chinese translation based on the best practices of international journals following the ARRIVE 2.0 guidelines in international journals, specifically for the complete interpretation of the ARRIVE 2.0 guidelines published in the PLoS Biology journal in 2020 (original text can be found at https://arriveguidelines.org). The fourth part of the article includes the items 1-5 of ARRIVE 2.0 Recommended 11 section, which covers "Abstract" "Background" "Objectives" "Ethical statement" and "Housing and husbandry". Its aim is to promote the full understanding and use of the ARRIVE 2.0 guidelines by domestic researchers, enhance the standardization of experimental animal research and reporting, and promote the high-quality development of experimental animal technology and comparative medicine research in China.

After implantation, complex and highly specialized molecular events render functionally distinct organ formation, whereas how the epigenome shapes organ-specific development remains to be fully elucidated. Here, nano-hmC-Seal, RNA bisulfite sequencing (RNA-BisSeq), and RNA sequencing (RNA-Seq) were performed, and the first multilayer landscapes of DNA 5-hydroxymethylcytosine (5hmC) and RNA 5-methylcytosine (m⁵C) epigenomes were obtained in the heart, kidney, liver, and lung of the human foetuses at 13-28 weeks with 123 samples in total. We identified 70,091 and 503 organ- and stage-specific differentially hydroxymethylated regions (DhMRs) and m⁵C-modified mRNAs, respectively. The key transcription factors (TFs), T-box transcription factor 20 (TBX20), paired box 8 (PAX8), krueppel-like factor 1 (KLF1), transcription factor 21 (TCF21), and CCAAT enhancer binding protein beta (CEBPB), specifically contribute to the formation of distinct organs at different stages. Additionally, 5hmC-enriched Alu elements may participate in the regulation of expression of TF-targeted genes. Our integrated studies reveal a putative essential link between DNA modification and RNA methylation, and illustrate the epigenetic maps during human foetal organogenesis, which provide a foundation for for an in-depth understanding of the epigenetic mechanisms underlying early development and birth defects.
Humans ; DNA Methylation ; RNA ; Epigenesis, Genetic ; DNA/genetics* ; Basic Helix-Loop-Helix Transcription Factors/genetics*

Objective:To investigate the clinical result of biological mesh and synthetic mesh in the repair of hiatal hernia.Methods:a prospective cohort study was conducted to collect and analyze the clinical data of 60 patients with hiatal hernia who were treated at Department of Hernia and Abdominal Wall Surgery, Beijing Chaoyang Hospital, Capital Medical University from May 2019 to Jan 2020. Intraoperative blood loss, hospital stay, clinical symptoms (heartburn, acid regurgitation, belching, early satiety, chest pain), VAS score, postoperative recurrence rate and complications were evaluated.Results:There was no significant difference in the overall repair effect between biological mesh group and synthetic mesh group ( P>0.05). All of the 60 patients underwent successful laparoscopic hiatal hernia repair and fundoplication. There were no massive bleeding caused by organ or vascular injury, and no peri-operative death. No recurrence of hiatal hernia, massive hemorrhage, pneumothorax, pleural effusion, gastrointestinal fistula, mediastinal infection or abscess were found during the follow-up of 6 months. Conclusion:There is no significant difference in short-term clinical effect between the use of biological mesh and synthetic mesh after hiatal hernia repair.

Objective To investigate the effect of hydrogen-rich saline postconditioning on the expression of vascular endothelial growth factor receptor-1 (VEGFR1) during myocardial ischemiareperfusion (I/R) injury in rats.Methods Thirty-six adult male Sprague-Dawley rats,weighing 250-280 g,were randomly divided into 3 groups (n=12 each) using a random number table:sham operation group (group S),group I/R and hydrogen-rich saline group (group H2).Myocardial I/R was induced by 30 min ligation of anterior descending branch of the left coronary artery followed by 2 h reperfusion.In group H2,hydrogen-rich saline 5 ml/kg was injected intravenously at 5 min before reperfusion,while the equal volume of normal saline was given in group I/R.Left ventricular systolic pressure (LVSP),left ventricular enddiastolic pressure (LVEDP) and ± dp/dtmax were measured and recorded during reperfusion and at 120 min of reperfusion.The rats were sacrificed at 120 min of reperfusion,and myocardial specimens were obtained for determination of myocardial infarct size,contents of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) (using ELISA),and expression of VEGFR1 (by Western blot).At 120 min of reperfusion,blood samples were collected from the common carotid artery to measure cardiac troponin I (cTnI) concentrations in serum.Results Compared with group S,LVSP and ± dp/dtmax were significantly decreased,LVEDP was increased,the myocardial infarct size was enlarged,cTnI concentrations in serum and contents of IL-6 and TNF-α were increased,and the expression of VEGFR1 was down-regulated at 120 min of reperfusion in H2 and I/R groups.Compared with group I/R,LVSP and ± dp/dtmax were significantly increased,LVEDP and myocardial infarct size were decreased,cTnI concentrations in serum and contents of IL-6 and TNF-α were decreased,and the expression of VEGFR1 was up-regulated at 120 min of reperfusion in group H2.Conclusion Hydrogen-rich saline postconditioning can reduce myocardial I/R injury possibly by upregulating myocardial VEGFR1 expression and inhibiting inflammatory responses in the myocardium of rats.

Background It is estimated that discoidin domain receptor 2 (DDR2) and matrix metalloproteinase-13 (MMP-13) play an important role in the development of tumor angiogenesis.However,their effects on choroidal neovascularizaiton (CNV) have not been clarified yet.Objective This study was to observe the expression pattern of DDR2 and MMP-13 in CNV and to further investigate the regulation role of MEK/ERK and PI3K/Akt pathways to the expression of DDR2 and MMP-13 in CNV.Methods CNV models were established in 78 Brown Norway (BN) rats by retinal photocoagulation with 532 nm laser.Then the animals were randomly divided into the normal control group (n =7),the model control group (n =39),PD98059 (MEK1 inhibitor) group (n =16) and LY294002 (PI3K inhibitor) group (n =16),and 5 mmol/L PD98059 or 5 mmol/L LY294002 3 μl was intravitreally injected 1 day and 7 days after photocoagulation in the PD98059 group or LY294002 group.The expression of DDR2 and MMP-13 mRNA and proteins in the CNV area were detected by using reverse transcription PCR (RT-PCR),and the expression levels of p-ERK/ERK and p-Akt/Akt protein were detected by Western blot assay.CNV thickness was measured by pathological examination 14 days after photocoagulation,and the changes of CNV thickness,the expression levels of DDR2 and MMP-13 in CNV were compared among the model control group,PD98059 group and LY294002 group.Results Three days after photocoagulation,the cells within the lasered lesions proliferated,then CNV formed 7 days after photocoagulation and became stable 14 days after photocoagulation.Immunohistochemistry staining indicated that DDR2 was weakly expressed in the cells of ganglion cell layer,inner nuclear layer and vascular endothelial cells;while MMP-13 was strongly expressed in the cells of the inner limiting membrane layer,photoreceptor layer and sclera layer.Both DDR2 and MMP-13 were strongly expressed in CNV area.Double immunofluorescence staining revealed that MMP-13 and DDR2 co-expression in CNV area.RT-PCR revealed that the relative DDR2 mRNA levels at 1 day,3 days,7 days and 14 days after photocoagulation were 55.22±4.03,47.74±2.23,14.82±4.56 and 5.59±2.47 respectively,while the relative MMP-13 mRNA levels were 25.54±3.83,43.51±4.36,10.90±4.00 and 5.23±3.23 respectively.Compared with the normal control group,the expression of DDR2 and MMP-13 were significantly increased (all at P＜0.05).Immunofluorescence staining showed that the relative fluorescence unit (RFU) values at 1 day,3 days,7 days and 14 days after photocoagulation were 2.73±0.53,5.21±0.31 and 3.22±0.33 for DDR2 and 1.66±0.17,3.57±0.44,2.67±0.21 for MMP-13,respectively.The RFU values in the PD98059 group and LY294002 group 14 days after photocoagulation were 1.14±0.19,1.03±0.14 for DDR2 and 1.37±0.25,1.24±0.20 for MMP-13,respectively.Compared with the model control group,the differences were statistically significant (both at P＜0.05).Western blot results showed that,compared to the normal control group,the expression levels of p-ERK and p-Akt pretein increased at day 7 after photocoagulation (both at P＜0.05),and returned back to the baseline at day 14 after photocoagulation (both at P＞0.05).Both PD98059 and LY294002 treatment were able to attenuate the thickness of CNV to 57.21％ and 50.34％ at day 14 after photocoagulation and further decrease the expression levels of DDR2 and MMP-13 in CNV (all at P＜0.05).Conclusions The expressions of DDR2 and MMP-13 up-regulate in laser-induced CNV.MEK/ERK and PI3K/Akt pathways suppress the development of CNV by regulating the expression of DDR2 and MMP-13.

BACKGROUNDThe extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae has increasingly become a major contributor to nosocomial infections and can exhibit multiple antibiotic resistance. Previous studies have focused on the resistance genes in ESBL-producing strains, and the resistance-associated genetic environment of non-ESBL-producing strains has been ignored until now. Here, we investigated the occurrence and characteristics of non-ESBL-producing K. pneumoniae, which potentially carries unexpressed resistance genes.

METHODSK. pneumoniae strains were collected from five medical institutions in China from February 2010 to August 2013. The VITEK-2 ESBL detection system was used as a primary screen to identify the ESBL-producing phenotype, and the three primary types of ESBL-associated genes (CTX, SHV, and TEM) were detected by polymerase chain reaction (PCR) to confirm the strains presenting with a non-ESBL-producing phenotype. mRNA expression in the non-ESBL-producing strains was further screened by reverse-transcription PCR (RT-PCR) to validate their transcriptional efficiency.

RESULTSOut of 224 clinically isolated antibiotic-sensitive K. pneumoniae strains with a non-ESBL-producing phenotype, 5 (2.2%) were identified to carry inactivated ESBL blaSHV genes with intact upstream promoter regions and resistance gene sequences. Interestingly, three of the five antibiotic-sensitive K. pneumoniae strains containing ESBL blaSHV genes still exhibited mRNA transcription of blaSHV, while the other two exhibited no mRNA transcription.

CONCLUSIONThese findings suggest that inactivated ESBL genes exist in non-ESBL-producing antibiotic-sensitive K. pneumoniae strains, which have the potential to transform the strain into an ESBL phenotype if an inappropriate application or overdose of antibiotics is implemented during clinical management.

Anti-Bacterial Agents ; pharmacology ; China ; Drug Resistance, Multiple, Bacterial ; genetics ; Humans ; Klebsiella pneumoniae ; drug effects ; enzymology ; genetics ; Microbial Sensitivity Tests ; beta-Lactamases ; genetics

Objective To determine the dose-response relationship of 0.2％ ropivacaine for ultrasoundguided stellate ganglion block (SGB).Methods Seventy-five ASA physical status [or Ⅱ patients with migraine,aged 23-55 yr,with body mass index of 22-28 kg/m2,scheduled for elective ultrasound-guided SGB,were randomly divided into R1-5 groups (n =15 each) using a random number table.In R1,R2,R3,R4 and R5 groups,the patients underwent ultrasound-guided SGB with 0.2％ ropivacaine 1,2,3,4 and 5 ml,respectively.A successful SGB block was confirmed by the onset of ptosis (Horner syndrome) on the injected side.Probit analysis was used to calculate the effective dose of 0.2 ％ ropivacaine in 50 ％ and 95 ％ of the patients (ED50 and ED95) and 95％ confidence interval (95％ CI).Results The ED50 of 0.2％ ropivacaine for ultrasound-guided SGB was 2.2 ml (95％CI 1.9-2.5 ml) and ED95 was 3.2 ml (95％CI 2.8-4.1 ml).Conclusion The ED50 and ED95 of 0.2％ ropivacaine for ultrasound-guided SGB are 2.2 and 3.2 ml,respectively.

Background Our previous study demonstrated that hyperglycemia aggravate the choroidal neovascularization (CNV) by promoting the chemotaxis process of bone marrow-derived cells (BMCs).Bioluminescence imaging (BLI) can dynamically monitor CNV in vivo.However,how diabetes mellitus (DM)participate in CNV is still in research.Objective This study was to dynamically observe the influence of BMCs to CNV under hyperglycaemia by using BLI combined with histopathology.Methods BMCs from luciferase-green fluorescent protein (Fluc-GFP) double transgenic mice were injected to adult wild type C57BL/6J mice (nine mice per group) via caudal vein to create the chimera models with a chimerism degree higher than 85％,and the chimeric mice were randomized into the control group and DM group based on randomized number table.Streptozotocin [60 mg/(kg · d)] was intraperitoneally injected daily for 5 days to establish the DM models in the chimeric mice of the DM group.CNV was induced in the chimeric mice of both control group and DM group with 532 nm laser photocoagulation.BLI signal of BMCsFluc+GFP+ was in vivo examined by IVIS Kinetics system 1,3,5,7,14,21 and 28 days after CNV modeling.At the seventh day after laser,part of mice were sacrificed,and choroidal and retinal sections were prepared for histopathological examination.The length and thickness of CNV were compared between the control group and DM group.The use and care of experimental followed Statement of ARVO.Results The chimerism degree of the chimeric mice was (88.85 ± 2.46) ％ 28 days after BMCs transplantation,and the blood glucose concentration in the DM group was (17.88±0.86)mmol/L.Histopathological examination revealed that CNV broke through the Bruch membrane toward subretinas.The length of the CNV was (338.67±33.17) μm in the DM group and (180.33±24.68)μm in the control group,showing a significant difference between the two groups (t =8.943,P＜0.05).However,no significant difference was seen in the CNV thickness between the two groups (t =1.790,P＞0.05).Light signals appeared 1 day and reach strongest 7 days after CNV modeling in both groups.The Light signals were stronger in the DM group than those in the control group on 5,7,14 and 21 days after CNV modeling (t =3.411,5.594,5.067,2.663,all at P＜0.05).Conclusions Hyperglycemia can promote more BMCs to participate in the pathogenesis and aggravation of CNV.The behavior of BMCs in CNV can be evaluated using BLI in vivo.

OBJECTIVETo determine if locally administered bone morphogenetic protein-2 (BMP-2) and osteoprotegerin (OPG) improved osteogenesis and new bone formation by trans-sutural distraction osteogenesis.

METHODSTwenty four dogs were divided into three groups randomly and received new internal trans-sutural distraction osteogenesis treatment. Five days after operation, infusion apparatus with double-tube was inserted to submucosa near the distracted zone to deliver controlled release agent of recombinant human bone morphogenetic protein-2/poly (lactic-co-glycolic acid)/fibrin sealant (rhBMP-2/PLGA/FS) in group A and group C. Recombinant human osteoprotegerin/fibrin sealant (rhOPG/ FS) was injected three weeks later in group B and group C. Histology staining and bone histomorphometry were used to measure the changes of maxillary bone sutura after distraction for 1, 2, 4 and 6 weeks.

RESULTSNew bone formation observed in distracted zone showed a significant increase in group A and C. Transmission electron microscope showed the osteoblast and osteocyte were active with dilated rough endoplasmic reticulum and a large number of chondriosomes and Golgi complex. After distraction for 6 weeks, indexes of osteoblast of group A, B, and C were 38.5 +/- 7.7, 35.7 +/- 6.5, and 41.7 +/- 11.0, indexes of osteoclast (Ioc) were 5.9 +/- 1.0, 1.2 +/- 0.3, and 2.8 +/- 0.4, bone trabecula thicknesses were (38.36 +/- 13.28), (66.20 +/- 9.16), and (51.85 +/- 9.92) microm respectively. Increased bone density and decreased Ioc were found in group B and C.

CONCLUSIONThe new elastic distractor is effective in inducing new bone formation. BMP-2 and OPG combination acts synergistically, and leads to significant enhancement of bone formation and remodeling.

Animals ; Bone Density ; Bone Morphogenetic Protein 2 ; Dogs ; Humans ; Lactic Acid ; Maxilla ; Osteoblasts ; Osteogenesis ; Osteogenesis, Distraction ; Osteoprotegerin ; Polyesters ; Polyglycolic Acid ; Polymers ; Recombinant Proteins ; Transforming Growth Factor beta