Children and adults differ significantly in physiology,biochemistry and immune function,which leads to sig-nificant differences in blood transfusion strategies between children and adults.To guide the clinical transfusion practice of pediatric patients and improve the prognosis of children,the National Health Commission organized the formulation and re-lease of the health industry standard Guideline for Pediatric Transfusion(WS/T 795-2022).This paper will briefly introduce some concepts that help understand of the Standard and the preparation process of the Standard,and explain and interpret the preparation of the"scope","general provisions"and"factors to consider"of the Standard,hoping to contribute to the understanding and implementation of the Standard.

Objective To observe the clinical efficacy of neck-shoulder-waist-leg pain cataplasms combined with electroacupuncture in the treatment of acute cervical radiculopathy(CR)with qi stagnation and blood stasis syndrome;To evaluate the synergistic effect and safety of neck-shoulder-waist-leg pain cataplasms.Methods Totally 132 CR patients with qi stagnation and blood stasis type in acute stage were randomly divided into treatment group(66 cases)and control group(66 cases).The treatment group was treated with electroacupuncture combined with neck-shoulder-waist-leg pain cataplasms,and the control group was treated with electroacupuncture combined with placebo.Both groups were treated once a day for 5 days and then rested for 2 days for 3 weeks,and follow-up was performed at 1 and 2 weeks after treatment.Clinical efficacy of both groups was observed.The short-form McGill pain questionnaire(SF-MPQ)score,neck disability index(NDI)score,and the F-wave conduction velocity of the median nerve and ulnar nerve were compared before and after treatment and at follow-up.Results 5 and 7 cases were lost in the treatment group and the control group,respectively.The total effective rate of the treatment group was 91.80%(56/61),and the recovery rate was 36.07%(22/61).The total effective rate of the control group was 84.75%(50/59),and the recovery rate was 18.65%(11/59).The difference in recovery rates between the two groups was statistically significant(P<0.05).Compared with before treatment,the SF-MPQ score and NDI score in both groups improved significantly(P<0.05)after treatment and at 1 and 2 weeks of follow-up,and the F-wave conduction velocity of the median and ulnar nerves increased significantly(P<0.05)after treatment.Comparison between two groups at the same time points showed that the SF-MPQ score and NDI score of the treatment group were better than those of the control group(P<0.05)after treatment and at 1 and 2 weeks of follow-up,and the F-wave conduction velocity of the median and ulnar nerves was faster than that of the control group(P<0.05)after treatment.There were no serious adverse reactions in the two groups.Conclusion Neck-shoulder-waist-leg pain cataplasms combined with electroacupuncture can improve the clinical symptoms of CR with qi stagnation and blood stasis syndrome in acute stage.Electroacupuncture plus neck-shoulder-waist-leg pain cataplasms can speed up the recovery process of the disease,which has synergistic effect.

Increasing evidences suggest the important role of calcium homeostasis in hallmarks of cancer, but its function and regulatory network in metastasis remain unclear. A comprehensive investigation of key regulators in cancer metastasis is urgently needed. Transcriptome sequencing (RNA-seq) of primary esophageal squamous cell carcinoma (ESCC) and matched metastatic tissues and a series of gain/loss-of-function experiments identified potassium channel tetramerization domain containing 4 (KCTD4) as a driver of cancer metastasis. KCTD4 expression was found upregulated in metastatic ESCC. High KCTD4 expression is associated with poor prognosis in patients with ESCC and contributes to cancer metastasis in vitro and in vivo. Mechanistically, KCTD4 binds to CLIC1 and disrupts its dimerization, thus increasing intracellular Ca²⁺ level to enhance NFATc1-dependent fibronectin transcription. KCTD4-induced fibronectin secretion activates fibroblasts in a paracrine manner, which in turn promotes cancer cell invasion via MMP24 signaling as positive feedback. Furthermore, a lead compound K279-0738 significantly suppresses cancer metastasis by targeting the KCTD4‒CLIC1 interaction, providing a potential therapeutic strategy. Taken together, our study not only uncovers KCTD4 as a regulator of calcium homeostasis, but also reveals KCTD4/CLIC1-Ca²⁺-NFATc1-fibronectin signaling as a novel mechanism of cancer metastasis. These findings validate KCTD4 as a potential prognostic biomarker and therapeutic target for ESCC.

Objectives: . Branchio-oto syndrome (BOS) primarily manifests as hearing loss, preauricular pits, and branchial defects. EYA1 is the most common pathogenic gene, and splicing mutations account for a substantial proportion of cases. However, few studies have addressed the structural changes in the protein caused by splicing mutations and potential pathogenic factors, and several studies have shown that middle-ear surgery has limited effectiveness in improving hearing in these patients. BOS has also been relatively infrequently reported in the Chinese population. This study explored the genetic etiology in the family of a proband with BOS and provided clinical treatment to improve the patient’s hearing. Methods: . We collected detailed clinical features and peripheral blood samples from the patients and unaffected individuals within the family. Pathogenic mutations were identified by whole-exome sequencing and cosegregation analysis and classified according to the American College of Medical Genetics and Genomics guidelines. Alternative splicing was verified through a minigene assay. The predicted three-dimensional protein structure and biochemical experiments were used to investigate the pathogenicity of the mutation. The proband underwent middle-ear surgery and was followed up at 1 month and 6 months postoperatively to monitor auditory improvement. Results: . A novel heterozygous EYA1 splicing variant (c.1050+4 A>C) was identified and classified as pathogenic (PVS1(RNA), PM2, PP1). Skipping of exon 11 of the EYA1 pre-mRNA was confirmed using a minigene assay. This mutation may impair EYA1-SIX1 interactions, as shown by an immunoprecipitation assay. The EYA1-Mut protein exhibited cellular mislocalization and decreased protein expression in cytological experiments. Middle-ear surgery significantly improved hearing loss caused by bone-conduction abnormalities in the proband. Conclusion . We reported a novel splicing variant of EYA1 in a Chinese family with BOS and revealed the potential molecular pathogenic mechanism. The significant hearing improvement observed in the proband after middle-ear surgery provides a reference for auditory rehabilitation in similar patients.

OBJECTIVE To study the sp ectrum-effect relationship of anti-gastric ulcer effect of Shaoyao gancao decoction. METHODS Eleven batches of Shaoyao gancao decoction were prepared ；gastric ulcer model was established by anhydrous ethanol modeling method. Using Weikangling as positive control ，the effects of Shaoyao gancao decoction on the contents of defensive factors [nitric oxide （NO），epidermal growth factor （EGF），superoxide dismutase （SOD）] and attack factor [malondialdehyde （MDA）] in gastric tissue of model rats were investigated. HPLC fingerprints of 11 batches of Shaoyao gancao decoction were established and similarity evaluation was performed with Similarity Evaluation System of Traditional Chinese Medicine Chromatographic Fingerprint （2004A edition ）； common peaks were identified by comparing with mixed control. The spectrum-effect relationship of Shaoyao gancao decoction against gastric ulcer was analyzed based on the grey correlation analysis. RESULTS Eleven batches of Shaoyao ganyao decoction could significantly decrease the content of MDA in gastric tissue ，while increased the contents of NO ，EGF and SOD in gastric ulcer model rats （P＜0.01），and had a certain inhibitory effect on the gastric ulcer. There were 23 common peaks in chromatograms of 11 batches of samples ，and the similarity with the control fingerprint was not less than 0.9. By comparing with mixed control ，7 common peaks were identified ，namely gallic acid （peak 5），albiflorin（peak 9），paeoniflorin（peak 10），liquiritin（peak 12），isoliquiritin apioside （peak 14），isoliquiritoside（peak 15）， glycyrrhizic acid （peak 22）. The average correlation degree of 7 identified common peaks and pharmacodynamic indexes were greater than 0.6，of which peak 22（glycyrrhizic acid ），peak 10（paeoniflorin）and peak 12（liquiritin）had the largest correlation ， and their values were 0.807，0.772 and 0.770 respectively. RESULTS The anti-gastric ulcer effect of Shaoyao gancao decoction is the result of the synergistic effect of multiple components ，among which glycyrrhizic acid ，paeoniflorin and liquiritin may be the main pharmacodynamic components.

Enzalutamide (ENZ) is a second-generation androgen receptor (AR) antagonist used for the treatment of castration-resistant prostate cancer (CRPC) and reportedly prolongs survival time within a year of starting therapy. However, CRPC patients can develop ENZ resistance (ENZR), mainly driven by abnormal reactivation of AR signaling, involving increased expression of the full-length AR (ARfl) or dominantly active androgen receptor splice variant 7 (ARv7) and ARfl/ARv7 heterodimers. There is currently no efficient treatment for ENZR in CRPC. Herein, a small molecule LLU-206 was rationally designed based on the ENZ structure and exhibited potent inhibition of both ARfl and constitutively active ARv7 to inhibit PCa proliferation and suppress ENZR in CRPC. Mechanically, LLU-206 promoted ARfl/ARv7 protein degradation and decreased ARfl/ARv7 heterodimers through mouse double minute 2-mediated ubiquitination. Finally, LLU-206 exhibited favorable pharmacokinetic properties with poor permeability across the blood-brain barrier, leading to a lower prevalence of adverse effects, including seizure and neurotoxicity, than ENZ-based therapies. In a nutshell, our findings demonstrated that LLU-206 could effectively inhibit ARfl/ARv7-driven CRPC by dual-targeting of ARfl/ARv7 heterodimers and protein degradation, providing new insights for the design of new-generation AR inhibitors to overcome ARfl/ARv7-driven CRPC.

The coronavirus disease 2019 (COVID-19) pandemic is caused by infection with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is spread primary via respiratory droplets and infects the lungs. Currently widely used cell lines and animals are unable to accurately mimic human physiological conditions because of the abnormal status of cell lines (transformed or cancer cells) and species differences between animals and humans. Organoids are stem cell-derived self-organized three-dimensional culture in vitro and model the physiological conditions of natural organs. Here we showed that SARS-CoV-2 infected and extensively replicated in human embryonic stem cells (hESCs)-derived lung organoids, including airway and alveolar organoids which covered the complete infection and spread route for SARS-CoV-2 within lungs. The infected cells were ciliated, club, and alveolar type 2 (AT2) cells, which were sequentially located from the proximal to the distal airway and terminal alveoli, respectively. Additionally, RNA-seq revealed early cell response to virus infection including an unexpected downregulation of the metabolic processes, especially lipid metabolism, in addition to the well-known upregulation of immune response. Further, Remdesivir and a human neutralizing antibody potently inhibited SARS-CoV-2 replication in lung organoids. Therefore, human lung organoids can serve as a pathophysiological model to investigate the underlying mechanism of SARS-CoV-2 infection and to discover and test therapeutic drugs for COVID-19.
Adenosine Monophosphate/therapeutic use* ; Alanine/therapeutic use* ; Alveolar Epithelial Cells/virology* ; Antibodies, Neutralizing/therapeutic use* ; COVID-19/virology* ; Down-Regulation ; Drug Discovery ; Human Embryonic Stem Cells/metabolism* ; Humans ; Immunity ; Lipid Metabolism ; Lung/virology* ; RNA, Viral/metabolism* ; SARS-CoV-2/physiology* ; Virus Replication/drug effects*

Objective:To explore the association between dopamine-β-hydroxylase (DβH), norepinephrine transporter (NET) gene polymorphisms and panic disorder(PD).Methods:The structured clinical interview for the diagnostic and statistical manual of mental disorders fourth edition (DSM-Ⅳ) axis Ⅰ disorders was administered by trained clinical psychiatrist, 139 patients with PD(PD group) and 196 healthy controls(control group) were enrolled in the study.Single nucleotide polymorphism(SNP) genotyping was performed using an improved multiplex ligation detection reaction technique.SPSS 16.0 and PLINK softwares were used to compare the allele frequency and genotype distribution.Results:(1)Compared with control group, PD group carried more G allele(76.3% vs 68.4%) and fewer A allele(23.7% vs 31.6%) in NET rs5569, and the difference was significant(χ 2=4.986, OR=0.67, 95% CI: 0.47-0.95, P<0.05). However, the correlation was no longer significant after adjusting for Bonferroni’s multiple testing( P>0.05). (2)The additive model of NET rs5569 showed a association with PD ( OR=0.68, 95% CI: 0.48-0.96, P<0.05). And the recessive model of DβH rs1611114 showed a association with PD( OR=0.42, 95% CI: 0.18-0.96, P<0.05). However, these correlations were no longer significant after adjusting for Bonferroni's multiple testing( P>0.05). (3)No matter allele or genotype, there were no significant differences in DβH (rs129882, rs1611114, rs1611115) and NET (rs2242446, rs28386840) gene polymorphisms between panic disorder group and control group(all P>0.05). Conclusion:The present study indicates that there is no significant association of DβH and NET gene polymorphisms with PD.

Objective:To understand the current status of HIV self-testing reagent use in pre-exposure prophylaxis (PrEP) and related factors in men who have sex with men (MSM).Methods:From December 2018 to December 2019, "Gold data" online platform (www.jinshuju.com) was used to conduct multicenter PrEP studies in Shenyang, Beijing, Chongqing and Shenzhen of China.Results:A total of 1 222 MSM PrEP users were included in the multicenter study. The average age of the participants was (31.5±8.7) years, and the number of sexual partners in the past three months was 3 ( P 25, P 75:2,6). The proportions of those who did not use condoms in anal sex with fixed, casual and commercial partners were 62.7% (456/727), 56.3% (440/781) and 41.0% (16/39), respectively. Up to 74.5% (910/1 222) of participants had used HIV self-testing reagents, and the number of HIV self-testing during last year was 3 ( P 25, P 75:2,5). The multivariate logistic regression analysis indicated that compared with age group >40 years, those with education level of junior high school or below, those with psychological identity as female, event driven PrEP users, those never using new type drugs in past 3 months, the participants aged 18- years (a OR=2.06, 95% CI: 1.35-3.14), 26- years (a OR=2.72, 95% CI: 1.77-4.17), 31- years (a OR=1.76, 95% CI: 1.19-2.59), undergraduates (a OR=2.18, 95% CI: 1.35-3.49), graduate students and above (a OR=3.06, 95% CI: 1.69-5.54), those with psychological identity as male (a OR=3.22, 95% CI: 1.55-6.70), daily PrEP users (a OR=1.35, 95% CI: 1.03-1.78), and new type drug users in the past three months (a OR=1.72, 95% CI: 1.30-2.28) had higher proportions of HIV self-testing behaviors. Conclusions:The proportion of HIV self-testing in MSM PrEP users was high, while it was relatively low in older age group, event driven PrEP users and MSM never using new type drugs. To assess and improve the effectiveness and compliance of PrEPs, it is necessary to provide better HIV self-testing service for MSM with low HIV self-testing rate.

Objective:To fabricate biogenic gas vesicles (GVs)- polyethyleneimine (PEI)-marrow mesenchymal stem cells (BMSCs) and evaluate its potential on stem cell tracking with ultrasound imaging.Methods:GVs were cationized by PEI to fabricate GVs-PEI. The diameter and zeta potential of GVs-PEI were determined. GVs-PEI were co-incubated with BMSCs to obtain GVs-PEI-BMSCs and stem cell uptake was observed by fluorescence microscope. The cell viability of GVs-PEI-BMSCs was verified by cell counting kit-8 (CCK-8) assay were set. Ultrasound imaging was performed on 0, 2, 4 and 6 d in agarose phantom to evaluate ultrasound imaging capability of GVs-PEI-BMSCs group and BMSCs group in vitro. GVs-PEI-BMSCs and BMSCs were injected into quadriceps femoris of SD rats, and ultrasound imaging was performed on 0, 2, 4 and 6 d to evaluate the ultrasound imaging capability in vivo. One-way analysis of variance and independent-sample t test were used to analyze the data. Results:The diameter and zeta potential of GVs-PEI were (383.63±11.55) nm and (18.48±2.20) mV. Plenty of GVs-PEI were observed in GVs-PEI-BMSCs through microscope. When BMSCs were incubated with GVs-PEI in absorbance ( A) 500 nm of 0.5 and 1.0, there were no significant changes in the cell viability of GVs-PEI-BMSCs at 24, 48 and 72 h ( F values: 7.078-11.982, all P>0.05). Compared with BMSCs, GVs-PEI-BMSCs showed better ultrasound imaging capability in vitro in all time points with still significantly different signal at 6 d (634.29±10.78 vs 2 864.51±100.86; t=-121.86, P<0.001). The ultrasound imaging capability of GVs-PEI-BMSCs in vivo was much better than that of BMSCs at each time point with still significantly different signal at 6 d (2 108.02±217.96 vs 267.71±7.87; t=-121.39, P<0.001). Conclusion:GVs-PEI-BMSCs are successfully fabricated with the advantages of significant ultrasound imaging capability, long duration and safety, which provide a brand-new means for stem cells tracking in vivo.