Background and aims Cancer cachexia is prevalent in various cancers and is associated with chemotherapy toxicity. However, limited data exist on the relationship between cachexia and immune-related adverse events (irAEs). The aim of this study is to explore the correlation between cancer cachexia and irAEs and its possible mechanism. Methods A murine model of orthotopic hepatocellular carcinoma (HCC) with cachexia was developed to evaluate the impact of T-cell infiltration into multiple tumor-free organs on the occurrence of irAEs. Single-cell RNA sequencing of thymic stromal cells was performed. Additionally, patients with advanced cancers receiving anti-programmed cell death protein 1/ligand 1 (PD-1/L1) antibody treatment were followed to investigate the relationship between cachexia and irAEs. Results Inflammatory cell infiltration was observed in multiple tumor-free organs of cachexic HCC mice but not in non-cachexic controls. Immunofluorescence confirmed that the infiltrating cells included CD4+ and CD8+ T cells. Morphological assessment and hematoxylin–eosin staining revealed thymic atrophy in cachexic HCC mice. Single-cell RNA sequencing of thymic stromal cells showed a reduction in medullary thymic epithelial cells (mTECs) II and III in cachexic mice. Autoimmune regulator (Aire) downregulation was accompanied by decreased expression of tissue-restricted antigens in mTECs. T cells from cachexic HCC mice induced organ-specific inflammation and T-cell infiltration in multiple organs of tumor-free mice. Following anti-mouse PD-1 antibody treatment, the incidence of inflammation in multiple organs markedly increased in cachexic HCC mice and in tumor-free mice that had received T cells from the cachexic HCC mice. Flow cytometry and immunofluorescence analyses revealed enrichment of thymic monocytic myeloid-derived suppressor cells (M-MDSCs) in cachexic HCC mice. M-MDSCs infiltrated the thymus in cachexic mice with cancer, and they induced apoptosis of mTECs from tumor-free mice in vitro via nitric oxide production. Transfer of M-MDSCs led to inflammatory cell infiltration in multiple organs and thymic involution in tumor-free mice without affecting body weight. Sixty-four patients with advanced cancer receiving anti-PD-1/L1 therapy were enrolled. Patients who developed irAEs had higher levels of circulating M-MDSCs than those who did not. Moreover, patients with cachexia (body mass index (BMI) < 20 kg/m2 or ≥5% weight loss over the past 6 months) had elevated M-MDSC levels. Patients with both high M-MDSC levels and low BMI or weight loss ≥5% experienced more irAEs (hazard ratio: 2.333; 95% confidence interval: 1.231–4.423). Conclusions M-MDSCs in cachexic mice induced mTEC apoptosis through nitric oxide production, impairing T-cell negative selection and promoting autoimmune T-cell infiltration into tumor-free organs. Cancer cachexia-related M-MDSCs may serve as predictive biomarkers for irAEs in patients with advanced cancer. Graphical abstract

OBJECTIVES: To investigate the risk factors for neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture and establish a nomogram model for predicting the risk of neonatal asphyxia. METHODS: A retrospective study was conducted with 613 cases of neonatal asphyxia treated in 20 cooperative hospitals in Enshi Tujia and Miao Autonomous Prefecture from January to December 2019 as the asphyxia group, and 988 randomly selected non-asphyxia neonates born and admitted to the neonatology department of these hospitals during the same period as the control group. Univariate and multivariate analyses were used to identify risk factors for neonatal asphyxia. R software (4.2.2) was used to establish a nomogram model. Receiver operator characteristic curve, calibration curve, and decision curve analysis were used to assess the discrimination, calibration, and clinical usefulness of the model for predicting the risk of neonatal asphyxia, respectively. RESULTS: Multivariate logistic regression analysis showed that minority (Tujia), male sex, premature birth, congenital malformations, abnormal fetal position, intrauterine distress, maternal occupation as a farmer, education level below high school, fewer than 9 prenatal check-ups, threatened abortion, abnormal umbilical cord, abnormal amniotic fluid, placenta previa, abruptio placentae, emergency caesarean section, and assisted delivery were independent risk factors for neonatal asphyxia (P<0.05). The area under the curve of the model for predicting the risk of neonatal asphyxia based on these risk factors was 0.748 (95%CI: 0.723-0.772). The calibration curve indicated high accuracy of the model for predicting the risk of neonatal asphyxia. The decision curve analysis showed that the model could provide a higher net benefit for neonates at risk of asphyxia. CONCLUSIONS The risk factors for neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture are multifactorial, and the nomogram model based on these factors has good value in predicting the risk of neonatal asphyxia, which can help clinicians identify neonates at high risk of asphyxia early, and reduce the incidence of neonatal asphyxia.
Infant, Newborn ; Humans ; Male ; Pregnancy ; Female ; Nomograms ; Retrospective Studies ; Cesarean Section ; Risk Factors ; Asphyxia Neonatorum/etiology*

Objective: To analyze the clinical epidemiological characteristics including composition of pathogens , clinical characteristics, and disease prognosis acute bacterial meningitis (ABM) in Chinese children. Methods: A retrospective analysis was performed on the clinical and laboratory data of 1 610 children <15 years of age with ABM in 33 tertiary hospitals in China from January 2019 to December 2020. Patients were divided into different groups according to age,<28 days group, 28 days to <3 months group, 3 months to <1 year group, 1-<5 years of age group, 5-<15 years of age group; etiology confirmed group and clinically diagnosed group according to etiology diagnosis. Non-numeric variables were analyzed with the Chi-square test or Fisher's exact test, while non-normal distrituction numeric variables were compared with nonparametric test. Results: Among 1 610 children with ABM, 955 were male and 650 were female (5 cases were not provided with gender information), and the age of onset was 1.5 (0.5, 5.5) months. There were 588 cases age from <28 days, 462 cases age from 28 days to <3 months, 302 cases age from 3 months to <1 year of age group, 156 cases in the 1-<5 years of age and 101 cases in the 5-<15 years of age. The detection rates were 38.8% (95/245) and 31.5% (70/222) of Escherichia coli and 27.8% (68/245) and 35.1% (78/222) of Streptococcus agalactiae in infants younger than 28 days of age and 28 days to 3 months of age; the detection rates of Streptococcus pneumonia, Escherichia coli, and Streptococcus agalactiae were 34.3% (61/178), 14.0% (25/178) and 13.5% (24/178) in the 3 months of age to <1 year of age group; the dominant pathogens were Streptococcus pneumoniae and the detection rate were 67.9% (74/109) and 44.4% (16/36) in the 1-<5 years of age and 5-<15 years of age . There were 9.7% (19/195) strains of Escherichia coli producing ultra-broad-spectrum β-lactamases. The positive rates of cerebrospinal fluid (CSF) culture and blood culture were 32.2% (515/1 598) and 25.0% (400/1 598), while 38.2% (126/330)and 25.3% (21/83) in CSF metagenomics next generation sequencing and Streptococcus pneumoniae antigen detection. There were 4.3% (32/790) cases of which CSF white blood cell counts were normal in etiology confirmed group. Among 1 610 children with ABM, main intracranial imaging complications were subdural effusion and (or) empyema in 349 cases (21.7%), hydrocephalus in 233 cases (14.5%), brain abscess in 178 cases (11.1%), and other cerebrovascular diseases, including encephalomalacia, cerebral infarction, and encephalatrophy, in 174 cases (10.8%). Among the 166 cases (10.3%) with unfavorable outcome, 32 cases (2.0%) died among whom 24 cases died before 1 year of age, and 37 cases (2.3%) had recurrence among whom 25 cases had recurrence within 3 weeks. The incidences of subdural effusion and (or) empyema, brain abscess and ependymitis in the etiology confirmed group were significantly higher than those in the clinically diagnosed group (26.2% (207/790) vs. 17.3% (142/820), 13.0% (103/790) vs. 9.1% (75/820), 4.6% (36/790) vs. 2.7% (22/820), χ²=18.71, 6.20, 4.07, all P<0.05), but there was no significant difference in the unfavorable outcomes, mortility, and recurrence between these 2 groups (all P>0.05). Conclusions: The onset age of ABM in children is usually within 1 year of age, especially <3 months. The common pathogens in infants <3 months of age are Escherichia coli and Streptococcus agalactiae, and the dominant pathogen in infant ≥3 months is Streptococcus pneumoniae. Subdural effusion and (or) empyema and hydrocephalus are common complications. ABM should not be excluded even if CSF white blood cell counts is within normal range. Standardized bacteriological examination should be paid more attention to increase the pathogenic detection rate. Non-culture CSF detection methods may facilitate the pathogenic diagnosis.
Adolescent ; Brain Abscess ; Child ; Child, Preschool ; Escherichia coli ; Female ; Humans ; Hydrocephalus ; Infant ; Infant, Newborn ; Male ; Meningitis, Bacterial/epidemiology* ; Retrospective Studies ; Streptococcus agalactiae ; Streptococcus pneumoniae ; Subdural Effusion ; beta-Lactamases

To summarize the status quo of clinical evidence on oral Chinese patent medicine in the treatment of hyperlipidemia through scoping review and thereby provide a reference for clinical application and decision-making in health care. Proprietary Chinese medicines for the treatment of hyperlipidemia （dyslipidemia） were retrieved from relevant catalogs and then screened based on their instructions. Articles on the selected Chinese patent medicines were searched from Chinese and english electronic databases and screened according to the inclusion criteria， followed by data extraction and analysis. The results were described with text and graphs. ①A total of 32 Chinese patent medicines and 1 010 related articles were screened out. ②All the included Chinese patent medicines were made from Chinese medicinal materials， of which 5 were Chinese medicinal extracts，and the remaining 27 were pure Chinese medicinal preparations （the compositions of 1 prescription is confidential）. ③As indicated in the instructions， all the 32 Chinese patent medicines can be used to treat hyperlipidemia with the main syndrome of combined phlegm and blood stasis， and the main effect of them is activating blood and resolving stasis. ④Among the 32 medicines， Xuezhikang tablets （Capsules） and Gypenosides Tablets have the lowest price. ⑤For the 328 key clinical studies， 248 were randomized controlled trials and 80 non-randomized controlled trials. ⑥As for the sample size， randomized controlled trials generally included 28-579 cases， and non-randomized controlled trials 24-152 cases. Clinical studies with more than 200 cases accounted for 9.3% （12/129）. ⑦The most common method was the comparison of the intervention effect of Chinese patent medicine and western medicine （36.5%） and Atorvastatin Calcium Tablets was the preferred western medicine control. The observation duration was generally 56 days （8 weeks）. ⑧In clinical practice， the absolute value of blood lipid decrease and （total） effective rate were often used to evaluate the efficacy， and the effective rate was generally determined based on Clinical Guidelines for New Chinese Medicines. The overall quality of clinical studies on oral Chinese patent medicines is uneven and there is a lack of high-quality clinical evidence. Moreover，there are the risks of unreasonable use and uneasy use of Chinese patent medicine.It is recommended that researchers on proprietary Chinese medicine should further broaden the research ideas，focus on the top-level design of the research plan， and standardize the research process，thereby provide high-quality research evidence for the clinical use of proprietary Chinese medicine. Moreover， more efforts should be made to establish a sound mechanism for information collection and feedback of proprietary Chinese medicine， so as to reduce the risk of uneasy use.We hope that in-depth investigation and discussion should be launched by price， economy， quality evaluation， and other departments headed by the supervision department to make the price of Chinese patent medicine reasonable and fair， thereby promoting the rational use of Chinese patent medicine.

The aim of this paper was to obtain low toxicity and high efficiency anti-tumor Chinese medicine through screening the combination ratios of Momordicae Semen and Epimedii Folium, and to explore the anti-tumor mechanism of the combination of two drugs by observing their effect on apoptosis-related proteins in cancer cells. Methyl thiazolyl tetrazolium(MTT) assay was used to observe the effect of drug combination on the proliferation of tumor cells from different tissue sources. The effects of the combination of the two drugs on tumor cells were analyzed by Compusyn software. Plate cloning assay was used to observe the effect of combination of these two drugs on the proliferation of A549 cells in vitro. The expression of reactive oxygen species(ROS) and apoptotic proteins p53, Bcl-2 and Bax were compared by using ROS kit and Western blot. Lewis lung cancer model was used to observe the anti-tumor effect of drugs in vivo. The results showed that the anti-tumor effect of their ethanol extract was more significant than that of water extract, and the anti-proliferation effect was strongest when the ratio was 1∶1(P<0.05). Compusyn analysis showed that the combination of the two drugs had synergistic effect. Further studies showed that after combined use, the number of clonogen formation in A549 cells was significantly reduced(P<0.01); ROS production was increased; the expression of apoptosis-related protein p53 was up-regulated, and the ratio of Bcl-2/Bax was decreased. In vivo animal study showed that the tumor inhibition rate was 53.06%(P<0.05) in the high dose group. As compared with the single use of the two drugs, the combination of the two drugs had more significant anti-proliferative effect on tumors, and the optimum ratio was 1∶1. The combination of the two drugs at a ratio of 1∶1 inhibited the proliferation of various tumor cells, and had no significant effect on normal liver cells LO2 when compared with other ratios. Therefore, it can be preliminarily inferred that the combination of the two drugs may have the effect of synergism and detoxification. Further studies showed that the combination of the two drugs can significantly inhibit the proliferation of A549 cells, and its mechanism may be related to the activation of endogenous apoptotic pathway. In vivo experiments also showed that the tumor inhibition rate increased with the increase of drug concentration.
A549 Cells ; Animals ; Antineoplastic Agents, Phytogenic/pharmacology* ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Drugs, Chinese Herbal/pharmacology* ; Epimedium/chemistry* ; Humans ; Lung Neoplasms/drug therapy* ; Momordica/chemistry* ; Neoplasms, Experimental/drug therapy* ; Plant Leaves/chemistry*

OBJECTIVE: To screen and verify the differentially expressed genes related with aging of bone marrow mesenchymal stem cells (BM-MSCs) in acute myeloid leukemia (AML) patients by bioinformatics, so as to provide new molecular markers for the research and clinical treatment of AML. METHODS: The gene expression profiling chip related with BM-MSCs in AML patients in our hospital and the gene chip GSE84881 selected from NCBI database GEO were used for data analysis and exploration. The DAVID analysis software was used to perform gene ontology (GO) enrichment analysis and KEGG pathway enrichment analysis. Furthermore, the differentially expressed genes related with aging of BM-MSCs in AML patients were identified. Bone marrow samples were collected and MSCs were amplified in vitro, and RT-PCR was used to verify the differentially expressed genes, which should be further identified with senescence-associated β-galactosidase staining and MTT cell proliferation assays. RESULTS: A total of 247 differentially expressed genes were screened out by bioinformatics methods, including genes of 132 up-regulated expression and 115 down-regulated expression. Six differentially expressed genes related with aging of BM-MSCs in AML patients were screened out, including the genes of up-regulated expression, COL3A1 (P<0.05), CRYAB (P<0.01), DCN (P<0.05), and the genes of down-regulated expression, including CCL2 (P<0.05), CTSC (P<0.01) and IL6 (P<0.05). These 6 differentially expressed genes were consistent with data from chip assays, and which was significantly correlated with aging of BM-MSCs in AML patients. Meanwhile, the positive rate of senescence-associated β-galactosidase staining in BM-MSCs of AML patients was significantly different from that of healthy donors (P<0.01). MTT cell proliferation assay showed that BM-MSCs in AML patients had proliferative ability lower than the healthy donors' BM-MSCs. CONCLUSION The data here suggest novel clues for the clinical research and treatment of BM-MSCs aging in AML patients.
Bone Marrow Cells ; Cell Proliferation ; Cells, Cultured ; Computational Biology ; Humans ; Leukemia, Myeloid, Acute ; Mesenchymal Stem Cells

Objective: To explore the feasibility of free circumflex scapular artery perforator flap for repairing the wounds of the limbs in children. Methods: From April, 2010 to October, 2017. 39 cases of pediatric patients who suffered from skin and soft defects in the limbs with exposure of bone, joint or tendon were repaired by the circumflex scapular artery perforator flap.The flap size ranged from 6.0 cm×3.5 cm to 16 cm×14 cm. Doppler detection was used to determine the distribution of the descending, ascending and transverse branches of the circumflex scapular artery. The proper perforator flap type was selected according to the shape and the size of the wound. Results: Thirty-seven flaps survived smoothly. Venous crisis occurred in one case and arterial crisis occurred in another case at the second postoperative day. Both flaps survived completely after exploration. All flaps were healed at the recipient sites. Delayed healing of the donor site wound occurred in two cases. All cases were followed up for 3 to 36 months (average 12 months), and the appearance and function of the recipient sites were satisfactory. Only linear scars remained in the donor sites. Conclusions The free circumflex scapular artery perforator flap is an ideal method to resurface the soft-tissue defects of extremities in children.

Objective To observe the clinical efficacy and safety of the tiotropium bromide powder inhalation combined with theophylline sustained-release tablets in the treatment of chronic obstructive pulmonary disease(COPD) in stable phase.Methods Forty patients of COPD in stable phase were randomly divided into control group and treatment group with 20 cases per group.Control group was given theophylline sustained-release tablets 0.2 g,bid,oral.Treatment group was given tiotropium bromide powder inhalation 18 μg,qd,Inhalation therapy.The clinical efficacy,mean blood oxygen saturation at nigh (MSaO2),lowest blood oxygen saturation (LSaO2),the ratio on the blood oxygen saturation ＜ 90％ of the total amount of sleep time (T90) and adverse drug reactions were compared between two groups.Results After treatment,the total effective rates in the treatment and control groups were 95.00％ (19 cases/20 cases)and 65.00％ (13 cases/20 cases) with statistically significant difference (P ＜ 0.05).After treatment,the main indexes in the treatment and control groups were compared:MSaO2 were (93.72 ± 3.44) ％ and (86.42 ± 4.11) ％,LSaO2 were (92.53 ±5.64)％ and(86.94 ±6.23)％,T90 were(28.14 ±6.31)％ and(31.22 ±7.81)％,the differences were statistically significant(all P ＜0.05).The adverse drug reactions in treatment group were based on dry mouth,which in control group were based on mild stomach discomfort.The incidences of adverse drug reaction in the treatment and control groups were 10.00％ and 5.00％ without significant difference(P ＞ 0.05).Conclusion Tiotropium bromide powder inhalation combined with theophylline sustained-release tablet has a definitive clinical efficacy and safety in the treatment of COPD in stable phase,which can improve the respiratory function.

The study was aimed to investigate the effect of CIAPIN1 gene on the proliferation of chronic myeloid leukemia (CML) cell line K562. The shRNA eukaryotic expression vector targeting CIAPIN1 gene was constructed and transfected into K562 cells. The inhibitory efficiency on K562 cells was detected by real-time PCR and Western blot; the proliferative activity of K562 cells was detected by MTT assay; the number and size of colonies were assessed by using colony-forming test; the tumorigenic potential was tested in vivo by using nude mice. The results indicated that as compared with control group, the CIAPIN1 gene expression statistically decreased; the proliferative activity of K562 cells in interference group was distinctly weakened; the number and size of colonies were significantly reduced; the tumorigenic potential was also lowered in vivo. It is concluded that inhibition of CIAPIN1 expression can inhibit K562 cell proliferation in vitro and in vivo.
Cell Proliferation ; Genetic Vectors ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; RNA, Small Interfering ; Transfection

Hematopoietic stem cells are capable of self-renewal or differentiation when they divide. Three types of cell divisions exist. A dividing stem cell may generate 2 new stem cells (symmetrical renewal division), or 2 differentiating cells (symmetrical differentiation division), or 1 cell of each type (asymmetrical division). This study was aimed to explore an efficient and stable method to distinguish the way of cell division in hematopoietic stem cells. Previous studies showed that the distribution of Numb in a cell could be used to distinguish the type of cell division in various kinds of cells. Therefore, the distribution of Numb protein was detected by immunofluorescence in mitotic CD48(-)CD150(+)LSK cells of mice exploring the relationship between Numb protein and centrosomes. Since CD48 positive marks the HSC that have lost the ability to reconstitute the blood system in mice, CD48 marker could be used to distinguish cell fate decision between self-renewal and differentiation as a living marker. In this study, the CD48(-)CD150(+)LSK cells were sorted from bone marrow cells of mice and the cells were directly labeled with Alexa Fluor (AF) 488-conjugated anti-CD48 antibody in living cultures. After 3 days, the percentage of AF488(+) cells was evaluated under microscope and by FACS. Then colony forming cell assay (CFC) was performed and the ability of cell proliferation were compared between AF488(+) and AF488(-) cells. The results showed that Numb could be used to distinguish different cell division types of hematopoietic stem cells, which was symmetrically or asymmetrically segregated in mitotic CD48(-)CD150(+)LSK cells. The self-labeled fluorochrome could be detected both by FACS as well as microscope. There were about 40% AF488(+) cells after 3 day-cultures in medium titrated with self-labeled AF 488-conjugated anti-CD48 antibody, and the results were consistent between confocal fluorescence microscopy and flow cytometry analysis. The colony forming ability of AF488(+) cells was significantly higher than that of AF488(-) cells (P < 0.05). The proliferation ability of AF488(-) cells was also significantly higher than AF488(+) cells (P < 0.05). It is concluded that the expression of CD48 can distinguish cell division of hematopoietic stem cells and can be used as a live marker for the loss of stemness. In comparison with the Numb protein staining, this method can be used in living cells, thus provides greater convenience for subsequent cell culture studies and cell transplantation experiments.
Animals ; Antigens, CD ; metabolism ; Biomarkers ; metabolism ; CD48 Antigen ; Cell Division ; Cells, Cultured ; Hematopoietic Stem Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL