Objective To investigate the extracellular and intracellular free amino acid concentrations in healthy subjects and its changes in chronic renal failure(Car) .Methods The free amino acid concentrations in the plasma, in the erythrocytes and in the skeletal muscle of 15 patients with CRF were compared with those in ten healthy subjects.Results In healthy subjects, there was little ~difference between the concentrations in erythrocytes and in plasma for most amino acids; but in ~erythrocytes the concentrations of Asp, Glu, Ser were higher than those in plasma (P

Objective:To evaluate the relationship between the mechanism of promoting wound healing by modified autologous blood transfusion and metastasis-associated lung adenocarcinoma transcript 1 ( MALAT1) in diabetic mice. Methods:Twenty SPF ICR mice, weighing 21-25 g, in which the diabetic model was successfully established, were divided into 2 groups ( n=10 each) using a random number table method: modified preservation group (group I) and ordinary preservation group (group O). Peripheral venous blood samples were collected and stored in the corresponding preservation solution for 7 days.The platelet aggregation rate, blood glucose, serum glycosylated hemoglobin (GHB) and phosphodiesterase (DPG) concentrations and WBC were measured.Autologous blood was transfused back immediately after the wound model was established.The percentage of wound healing area was calculated at 7, 10 and 14 days after autologous blood transfusion.The expression of hypoxia-inducible factor-1α, vascular endothelial growth factor, matrix metalloproteinase-9, β-actin, type Ⅰ collagen (Col Ⅰ), Col Ⅲ protein and mRNA and MALAT1 was determined by Western blot, immunohistochemistry and quantitative real-time polymerase chain reaction respectively, at 14 days after transfusion. Results:Compared with group O, the blood glucose, serum concentrations of GHB and DPG, and WBC were significantly decreased, platelet aggregation rate was increased, the percentage of wound healing area was increased, the positive staining rate of Col Ⅰ and Col Ⅲ was increased, and the expression of hypoxia-inducible factor-1α, vascular endothelial growth factor, matrix metalloproteinase-9, ColⅠ, Col Ⅲ and β-actin protein and mRNA and MALAT1 was up-regulated in group I ( P<0.05). Conclusion:The mechanism by which modified autologous blood transfusion promotes wound healing may be related to up-regulating MALAT1 expression in diabetic mice.

Objective To develop and optimize a new method to extract miRNAs from plasma.Methods miRNAs were extracted from plasma by mixing it with the extraction solution that contained surfactant and by heating .Then the ribonuclease inhibitor was added into the extraction to prevent RNAs from degradation .The expression level of each miRNA was detected by real-time quantitative PCR in oder to evaluate the feasibility of this method .Results A method which extracted miRNAs from plamsa in just one step was established .The specificity , reproducibility and stability of this method have been demonstrated by real-time quantitative PCR .Conclusion The one-step method is simple , inexpensive , and plasma-saving.It seems like a new method for clinical examination of miRNAs from plasma .

Objective To study the occurrence of acute gastrointestinal injury under different critical status in Pediatric Intensive Care Unit,and to assess the connection between citrulline and acute gastrointestinal injury.To ana-lyze the correlation among serum citrulline and C-reaction protein(CRP),procalcitonin(PCT),creatinine,urea nitro-gen,bilirubin,lactate,albumin.To study the role of citrulline in evaluating the children condition,prognosis and changes in critically ill children.Methods Ninety-six pediatric patients who were critically ill at the Pediatric Intensive Care Unit of the Second Hospital of Lanzhou University were selected,excluding those with primary gastrointestinal diseases and chronic kidney disease.Clinical features were recorded and CRP,PCT,creatinine,urea nitrogen,bilirubin,lactate, and albumin were measured.Serum citrulline concentrations were measured by high performance liquid chromatography in 65 patients with gastrointestinal injury,and sensitivity,specificity and area under the receiver operating characteristic (ROC)curve were calculated.Results The incidence of acute gastrointestinal injuries in non-critical group,critical group,reorganization critical group were 47.06%(16/34 cases),75.00%(36/48 cases),92.86%(13/14 cases) (χ2=11.848,P=0.003).The serum citrulline concentrations of gastrointestinal injury group and non- gastrointesti-nal injury group were(14.655 ± 8.231)μmol/L,(23.522 ± 11.079)μmol/L(t=4.398,P<0.05).The area under ROC of citrulline was 0.079 8.Serum citrulline concentrations increased significantly in infants with very severe gastro-intestinal injury after 3 days of treatment compared with admission[(26. 40 ± 16. 10)μmol/L vs.(14. 40 ± 6.82)μmol/L,t=5.524,P=0.029].The lower serum citrulline levels the children had,the higher mortality was. There were negative correlations among serum citrullin and CRP,PCT and length of hospital stay(r = -0. 319,-0.299,-0.364,P=0.003,0.006,0.001).There were no correlations among serum citrullin and blood urea nitro-gen,lactic acid,albumin,and bilirubin levels.Conclusion The more critically ill children have a higher incidence of acute gastrointestinal injury and the worse the prognosis.Serum citrulline levels may well reflect the extent of acute gas-trointestinal injury and the outcome.Perhaps it can be one of the reliable markers to evaluate the function of gastrointes-tinal and participate in assessing critical condition and prognosis in critical ill children ness.

Background and objective 50%-70%of patients with advanced lung cancer will develop bone metas-tases. hTe aim of this study is to establish the nude mice bone metastasis model of lung adenocarcinoma using A549, H1299, SPC-A-1 and XL-2, all of which own different invasion and migration abilities in vitro and supervise the bone metastases by MicroCT. Methods iftfy BALB/C-nu/nu nude mice were grouped into ifve groups on average randomly. Cells of the four cell lines were injected into the letf cardiac ventricle of mice in the four experimental groups (0.2 mL/mouse) respectively;meanwhile, mice in the control group were injected with normal saline (0.2 mL/mouse) in the same manner. Periodical radio-logical examination was carried out to supervise the variation of the mice since the second week atfer injection. When mice in each group became thin obviously, end the experiment of this group. Before the end, pathological sections of bone tissues were made. We classiifed the bone metastatic sites into axial skeleton and limb bone, in order to compare the metastatic rates of these two different parts. hTe bone metastatic abilities of the four cell lines was statistically analyzed by comparing the average time cost in the appearance of bone metastases and the percentage of bone metastases among the experimental groups. Results Different metastatic sites which had been identiifed both by MicroCT and pathological sections appeared in each group of the four experimental groups. By contrast, no metastasis was observed in the control group. hTe percentage of cancer metastasiz-ing to axial skeleton was remarkably higher than the percentage of tumor metastasizing to the limb bone in each experimental group, which was consistent with the clinical regularity and characteristics of skeletal metastases with lung cancer. hTus, the model has been established triumphantly. However, there were no statistical differences in the average time consumed and skeletal metastatic rate among the four experimental groups. Conclusion hTe disruption in the bone can be clearly detected by MicroCT, which is beneift to supervise the osseous metastasis. We successfully developed the nude mice bone metastasis model of lung adenocarcinoma, which will pave the way for exploring novel prevention and therapy strategies clinically. hTe four cell lines varied in invasion and migration abilities in vitro, but there was no statistical difference in the metastatic ability in vivo, and the reason need to be explored further in future.

【Objective】 To explore the clinical features and treatment outcomes of female urethral carcinoma so as to improve the awareness and prognosis of this rare malignant disease. 【Methods】 Clinical data of 8 cases of female urethral carcinoma treated during Jan. 2012 to Dec.2022 at the Department of Urology of Peking University People’s Hospital were retrospectively analyzed. The patients underwent urodynamic tests, cystourethroscopy and pathological biopsy to confirm the diagnosis. Traditional radical surgery was performed in 5 cases, and radical surgery for lower urethral cancer with bladder preservation was performed in 3 cases. 【Results】 The patients aged 36 to 68 years, with a mean of 53.75 years. Urinary obstruction, lower urinary tract symptoms and urethral masses were common manifestations. Urodynamic tests indicated bladder outlet obstruction. After surgical treatment, radical surgery for lower urethral cancer with bladder preservation showed advantages over traditional radical surgery in terms of intraoperative bleeding, operation time and postoperative hospital stay. 【Conclusion】 Female primary urethral carcinoma is rare but invasive. Early diagnosis and radical surgery are crucial for improving the prognosis. Radical surgery for lower urethral cancer with bladder preservation has better treatment outcomes and postoperative quality of life compared to traditional radical surgery. For such patients, symptoms should be closely monitored and timely diagnosis and treatment should be performed.

Anti-virulence strategy has been considered as one of the most promising approaches to combat drug-resistant bacterial infections. Pore-forming toxins (PFTs) are the largest class of bacterial toxins, inflicting their virulence effect through creating pores on the cell membrane. However, current solutions for eliminating PFTs are mostly designed based on their molecular structure, requiring customized design for different interactions. In the present study, we employed erythroliposome (denoted as RM-PL), a biomimetic platform constructed by artificial lipid membranes and natural erythrocyte membranes, to neutralize different hemolytic PFTs regardless of their molecular structure. When tested with model PFTs, including α-hemolysin, listeriolysin O, and streptolysin O, RM-PL could completely inhibit toxin-induced hemolysis in a concentration-dependent manner. In vivo studies further confirmed that RM-PL could efficiently neutralize various toxins and save animals' lives without causing damage to organs or tissues. In addition, we explored the underlying mechanisms of this efficient detoxification ability and found that it was mainly macrophages in the spleen and the liver that took up RM-PL-absorbed toxins through a variety of endocytosis pathways and digested them in lysosomes. In summary, the biomimetic RM-PL presented a promising system for broad-spectrum and powerful toxin neutralization with a mechanism of lysosome-mediated toxin degradation.

ObjectiveTo explore the changes in volatile components， total polysaccharides， enzyme activity， and chromaticity value of Myristicae Semen Koji（MSK） during the fermentation process， and conduct correlation analysis. MethodsBased on gas chromatography-mass spectrometry（GC-MS）， the changes of volatile components in MSK at different fermentation times were identified. The phenol sulfuric acid method， dinitrosalicylic acid method（DNS）， and carboxymethyl cellulose sodium salt method（CMC-Na） were used to investigate the total polysaccharide content， amylase activity， and cellulase activity during the fermentation process. Visual analysis technology was used to explore the changes in chromaticity values， revealing the fermentation process of MSK and the dynamic changes of various measurement indicators， partial least squares-discriminant analysis（PLS-DA） was used to explore the differential compounds of MSK at different fermentation degrees， and Pearson correlation analysis was used to explore the correlation between volatile components of MSK and total polysaccharides， enzyme activity， and chromaticity values. ResultsA total of 60 volatile compounds were identified from MSK， the relative contents of components such as （+）-α-pinene， β-phellandrene， β-pinene， （+）-limonene， and p-cymene obviously increased， while the relative contents of components such as safrole， methyl isoeugenol， methyleugenol， myristicin， and elemicin significantly decreased. During the fermentation process， the total polysaccharide content showed an upward trend， while the activities of amylase and cellulase showed an initial increase followed by a decrease， and reached their maximum value at 40 h. the overall brightness（L^*） and total color difference（ΔE^*） gradually increased， while the changes in red-green value（a^*） and yellow-blue value（b^*） were not obvious. PLS-DA results showed that MSK could be clearly distinguished at different fermentation times， and 13 differential biomarkers were screened out. Pearson correlation analysis results showed that the contents of α-terpinene， β-phellandrene， methyleugenol， β-cubebene and myristic acid had an obvious correlation with chromaticity values. ConclusionAfter fermentation， the volatile components， total polysaccharides， amylase activity， and cellulase activity of MSK undergo significant changes， and there is a clear correlation between them and chromaticity values， which reveals the dynamic changes in the fermentation process and related indicators of MSK， laying a foundation for the quality control.

Human pluripotent stem cells provide an inexhaustible model to study human embryogenesis in vitro. Recent studies have provided diverse models to generate human blastoids by self-organization of different pluripotent stem cells or somatic reprogramming intermediates. However, whether blastoids can be generated from other cell types or whether they can recapitulate postimplantation development in vitro is unknown. Here, we develop a strategy to generate human blastoids from heterogeneous intermediates with epiblast, trophectoderm, and primitive endoderm signatures of the primed-to-naïve conversion process, which resemble natural blastocysts in morphological architecture, composition of cell lineages, transcriptome, and lineage differentiation potential. In addition, these blastoids reflect many features of human peri-implantation and pregastrulation development when further cultured in an in vitro 3D culture system. In summary, our study provides an alternative strategy to generate human blastoids and offers insights into human early embryogenesis by modeling peri- and postimplantation development in vitro.
Humans ; Pluripotent Stem Cells/metabolism* ; Embryo, Mammalian/metabolism* ; Cell Differentiation ; Blastocyst ; Cell Lineage ; Embryonic Development