This case report presents a 16-year-old male patient with deficiency of adenosine deaminase 2(DADA2). The patient had a history of Raynaud′s phenomenon with digital ulcers since childhood. As the disease progressed, the patient developed retinal vasculitis, intracranial hemorrhage, skin necrosis, severe malnutrition, refractory hypertension, and gastrointestinal bleeding. Genetic testing revealed compound heterozygous mutations in the ADA2 gene (paternal c.1072G > A pathogenic mutation + maternal c.1065C > A variant of unknown clinical significance). ADA2 enzyme activity was significantly reduced (0.1 U/L). A multidisciplinary treatment team developed an individualized plan to address key issues, including nutritional support, blood pressure control, rehabilitation, pain management, and balancing anticoagulation/bleeding risks. After systematic intervention, the patient′s condition showed partial improvement. This case reveals clinical challenges such as anticoagulation decisions and nutritional support of DADA2. It also emphasizes the importance of early molecular diagnosis, tumor necrosis factor-α inhibitor targeted therapy, and multidisciplinary collaboration in management, underlining the core value of precision medicine in rare disease management.

Objective: To explore the differences in the gut microbiota of primary school students with different levels of sugar sweetened beverage intake, so as to provide scientific evidence for better identification of health risks in children and the development of targeted health policies. Methods: In June 2022, a total of 192 healthy primary school students from Chengdu were selected using a stratified cluster random sampling method. The sugar sweetened beverage intake was assessed through a dietary frequency questionnaire. Based on the median daily sugar sweetened beverage intake, primary school students were categorized into a low intake group ( n =96) and a high intake group ( n =96). The gut microbiota in fresh fecal samples from the two groups of primary school students was analyzed using 16S rRNA high throughput sequencing, and the diversity and community structure differences in the gut microbiota were compared. Results: Children in the low intake group had a sugar sweetened beverage intake of (21.3±1.6) mL/d, while the high intake group had an intake of (269.6±37.3) mL/d. Diversity analysis results showed that there were no statistically significant differences between the low intake and the high intake group in terms of α diversity metrics: Observed_otus index [298.50 (259.75, 342.25), 305.50 (244.25, 367.75)], Goods_coverage index [1.00 (1.00, 1.00), 1.00 (1.00, 1.00)], Chao index [304.18 (260.75, 348.78), 305.88 (245.68, 370.88)], Shannon index [5.88 (5.29, 6.45), 5.71 (4.89, 6.28)] and Simpson index [0.95 (0.91, 0.97), 0.94 (0.88, 0.97)] ( Z =-0.64, -0.76, -0.54, -1.76, -1.67, P >0.05). Furthermore, no statistically significant difference was observed in β diversity between the two groups ( R 2=0.006, P >0.05). At the genus level, the abundance of Blautia [0.033 (0.018, 0.055)] and Fusicatenibacter [0.009 (0.005, 0.015)] were higher in the low intake group compared to the high intake group [0.024 (0.013, 0.041),0.006 (0.003, 0.011)]and differences were statistically significant ( Z =-2.52, -2.81, P <0.05). LEfSe analysis highlighted intergroup differences primarily in Blautia, Fusicatenibacter and Sarcina( LDA= 3.56,3.12,3.53, P <0.05). Conclusions There is no significant difference in the diversity and overall structure of the gut microbiota in primary school students with different levels of sugar sweetened beverage intake. However, there are species variations at the genus level. The information can serve as a scientific basis for identifying health risks in primary school students and formulating targeted health strategies.

ObjectiveKey microRNAs （miRNAs） of colorectal adenoma （CRA） were identified and analyzed by bioinformatics methods， and differentially expressed genes （DEGs） were screened to construct regulatory relationships. The mechanism of Xiezhuo Jiedu recipe in preventing CRA was speculated and verified by animal experiments. MethodThe miRNAs dataset GSE50194 was obtained from the Gene Expression Omnibus （GEO） database of intestinal mucosal tissue of CRA patients， and the differentially expressed miRNAs were screened by GEO2R and Excel. TargetScan， miRTarbase， and miRDB databases were used to predict the target genes of the differentially expressed miRNAs， and an intersection was obtained. Key DEGs were screened through the STRING database and Cytoscape software， and the TRRUST database was used to predict downstream binding transcription factors （TFs）. The mRNA intersection was enriched by gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） in the Metascape database. DIANA TOOLS were applied to perform KEGG enrichment analysis of key miRNAs， and the key signaling pathways were selected for animal experiments. In animal experiments， the CRA mice model was established by using sodium glycan sulfate （DSS） drinking combined with intraperitoneal injection of azomethane oxide （AOM）， and Xiezhuo Jiedu recipe and aspirin were given by intragastric administration at the same time. The experiment lasted for nine weeks. The pathological changes in intestinal tissue were observed by hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of miR-34a-5p in adenoma tissue. Protein expression levels of phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， phosphoryl-PI3K （p-PI3K）， phosphoryl-Akt （p-Akt）， and B cell lymphoma （Bcl）-2 were detected by Western blot. The expression of Cyclin D1 （CCND1） was detected by immunohistochemistry （IHC）. In situ terminal transferase labeling （TUNEL） was used to detect apoptosis of adenoma tissue cells. ResultThe GEO database screened the GSE50194 dataset， and miR-34a-5p was selected as the research object from CRA and normal tissue. A total of 93 DEGs were selected. Among them， GO and KEGG enrichment analyses were closely related to biological processes such as transcriptional regulatory complex， RNA polymerase Ⅱ transcriptional regulatory complex， enzyme-linked receptor protein signaling pathway， and DNA-binding transcriptional activator activity， cancer pathway， PI3K/Akt pathway， etc. miR-34a-5p is mainly enriched in PI3K/Akt， cell cycle， and colorectal cancer pathways. Five key DEGs were screened out through the Matescape database， among which Bcl-2 and CCND1 were the key DEGs of miR-34a-5p. Further screening of the TFs of key DEGs revealed that E2F transcription factor 1 （E2F1） and tumor protein P53 （TP53） were the main TFs of Bcl-2 and CCND1. Animal experiments showed that Xiezhuo Jiedu recipe could effectively up-regulate mRNA level of miR-34a-5p， down-regulate the expression of PI3K， Akt， Bcl-2， p-PI3K， and p-Akt proteins in the intestinal tissue of CRA mice， down-regulate the positive expression rate of CCND1， and increase the apoptosis rate of intestinal epithelial cells. ConclusionIt is speculated that Xiezhuo Jiedu recipe may inhibit the abnormal proliferation and promote the apoptosis of intestinal epithelial cells in CRA mice by regulating the miR-34a-5p/PI3K/Akt signaling pathway， thus playing a role in the prevention of CRA.

Objective To understand the internal quality control(IQC)situation of routine biochemical projects in Guizhou Province from 2020 to 2022,and to use the analysis batch length sigma rule to select the appropriate IQC strategy for routine biochemical projects.Methods A sum of 41,41 and 52 laboratories were selected from the laboratories participating in the first external quality assessment(EQA)program of the routine biochemical project in the Guizhou Provincial Center for Clinical Laboratory from 2020 to 2022,which uniformly used two concentration levels of internal quality control products produced by Randox Company in the UK.The coefficient of variation(CV)was calculated based on their IQC information,and then these CV values were used to compare with imprecise performance specifications such as WS/T403-2012 standards.The IQC situation of routine biochemical projects of Guizhou Province in recent years was analyzed.Randomly 15 laboratories were selected each year to report the results of the triglyceride project,and their σ values were calculated.Based on the Westgard sigma rule flowcharts of the analysis batch length,appropriate quality control strategies were selected and the quality goal index(QGI)was calculated to evaluate analytical performance of laboratory detection system.Results In the current monthly CV,the proportion of laboratories that met the WS/T403-2012 standards for Ca and TP projects was relatively low,while the pass rates for 7 projects(P,CK,LDH and other projects)were all above 80%,showing an increasing trend year by year.The proportion of laboratories with the 7 projects(P,CK,LDH and other projects)meeting the performance specifications for biomedical variation(appropriate)all reached 90%,while the proportion of laboratories with 5 projects(Na,Ca,Cl and other projects)was relatively low.The proportion of 10 projects such as P,CK,and LDH in cumulative CV of laboratories that met the performance specifications for biomedical variation(appropriate)was over 90%.For the triglyceride project,from 2020 to 2022,there were 2,5 and 13 laboratories that reached 6 σ horizontal.Compared to values of 2020 and 2021 years,the trend of σ values changes of 2022 was increasing(t=3.855,3.511,P≤0.001),indicating that the analytical performance of these laboratories was getting better and better.Conclusion The IQC levels of routine biochemical projects in Guizhou Province were increasing year by year.The Westgard sigma rule of the analysis batch length is designed with personalized quality control rules for various laboratories in Guizhou Province,thus improving laboratory testing capabilities and better severing clinics.

Objective To construct plasmids and knock out HIF-1α gene expression in an naked mole rat skin fibroblasts(NSF)cell line using CRISPR/Cas9 genomic editing technology,to provide an in vitro cell model for studying the mechanism of hypoxia tolerance and the occurrence and development of hypoxia-related diseases in naked mole rats.Methods We designed four pairs of single guide RNA(sgRNA)sequences targeting exons 1～4 of the NSF HIF-1αgene and successfully constructed an expression plasmid.The plasmid with the optimal sgRNA was identified and transfected into 293T cells,and the supernatant was used for detecting the virus titer.Lentivirus particles carrying sgRNAs of HIF-1α were transfected into NSF cells which express Cas9 protein,based on a previous protocol.After transfection,fluorescence signals were observed under a fluorescence microscope,and HIF-1α expression in NSF cells was detected by Western Blot and T7 endonuclease 1(T7E1)analysis.Results Sanger sequencing showed that the designed sgRNA was successfully inserted into pX459 and pKLV2-U6-sgRNA2 vectors,demonstrating successful construction of a recombinant plasmid for transfection.T7E1 digestion successfully removed three bands and the target efficiency of sgRNA was 54％.Western Blot showed that the HIF-1α gene was successfully knocked out and its protein level was significantly reduced in NSF cells from naked mole rats(P=0.0019).There were no obvious morphological changes in HIF-1α-knockout cells under the microscope,and gene knockout had no obvious effect on cell proliferation.Conclusions We successfully constructed an HIF-1α-knockout cell line using CRISPR/Cas9 technology,to provide an experimental basis for further studies of the biological function of HIF-1α,as well as the mechanism of hypoxia tolerance in naked mole rats.The result also provide a theoretical foundation for the prevention and treatment of hypoxia-related diseases.

Objective:To investigate the relationship between intramuscular adipose tissue index (IATI) calculated from computed tomography images at transverse process of the first lumbar and all-cause mortality in maintenance dialysis patients, and to provide a reference for improving the prognosis in these patients.Methods:It was a multicenter retrospective cohort study. The clinical data of patients who received maintenance hemodialysis or peritoneal dialysis treatment from January 1, 2017 to December 31, 2019 in 4 grade Ⅲ hospitals including Zhongda Hospital Affiliated to Southeast University, Taizhou People's Hospital Affiliated to Nanjing Medical University, Affiliated Hospital of Yangzhou University, and the Third Affiliated Hospital of Soochow University were retrospectively collected. IATI was calculated by low attenuation muscle (LAM) density/skeletal muscle density. The receiver-operating characteristic curve was used to determine the optimal cut-off value of IATI, and the patients were divided into high IATI group and low IATI group according to the optimal cut-off value. The differences of baseline clinical data and measurement parameters of the first lumbar level between the two groups were compared. The follow-up ended on December 23, 2022. The endpoint event was defined as all-cause mortality within 3 years. Kaplan-Meier survival curve and log-rank test were used to analyze the survival rates and the differences between the two groups. Multivariate Cox regression analysis models were used to analyze the association between IATI and the risk of all-cause mortality in maintenance dialysis patients. Multivariate logistic regression analysis model was used to analyze the influencing factors of high IATI.Results:A total of 478 patients were eligibly recruited in this study, with age of (53.55±13.19) years old and 319 (66.7%) males, including 365 (76.4%) hemodialysis patients and 113 (23.6%) peritoneal dialysis patients. There were 376 (78.7%) patients in low IATI (<0.42) group and 102 (21.3%) patients in high IATI (≥0.42) group. The proportion of age ≥ 60 years old ( χ2=24.746, P<0.001), proportion of diabetes mellitus ( χ2=5.570, P=0.018), fasting blood glucose ( t=-2.145, P=0.032), LAM density ( t=-3.735, P<0.001), LAM index ( t=-7.072, P<0.001), and LAM area/skeletal muscle area ratio ( Z=-9.630, P<0.001) in high IATI group were all higher than those in low IATI group, while proportion of males ( χ2=11.116, P<0.001), serum albumin ( Z=2.708, P=0.007) and skeletal muscle density ( t=12.380, P<0.001) were lower than those in low IATI group. Kaplan-Meier survival analysis showed that the 3-years overall survival rate of low IATI group was significantly higher than that in high IATI group (Log-rank χ2=19.188, P<0.001). Multivariate Cox regression analysis showed that IATI<0.42 [<0.42/≥0.42, HR(95% CI): 0.50 (0.31-0.83), P=0.007] was an independent protective factor of all-cause mortality, and age ≥60 years old [ HR (95% CI): 2.61 (1.60-4.23), P<0.001], diabetes mellitus [ HR (95% CI): 1.71 (1.06-2.78), P=0.029] and high blood neutrophil/lymphocyte ratio [ HR (95% CI): 1.04 (1.00-1.07), P=0.049] were the independent risk factors of all-cause mortality in maintenance dialysis patients. Stepwise Cox regression analysis showed that IATI<0.42 was still an independent protective factor of all-cause mortality in maintenance dialysis patients [<0.42/≥0.42, HR (95% CI): 0.45 (0.27-0.76), P=0.003]. Multivariate logistic regression analysis showed that low skeletal muscle density [ OR (95% CI): 0.84 (0.81-0.88), P<0.001] and high serum triglyceride [ OR (95% CI): 1.39 (1.07-1.82), P=0.015] were the independent influencing factors of IATI≥0.42. Conclusion:IATI<0.42 of the first lumbar level is an independent protective factor of all-cause mortality in maintenance dialysis patients. Localized myosteatosis within high-quality skeletal muscle may reduce the risk of all-cause mortality in these patients.

ObjectiveTo investigate the effect and mechanism of modified Guizhi Fulingwan in preventing colorectal adenoma （CRA） in mice by regulating mitochondrial apoptosis pathway through the regulation of the phosphatase and tensin homolog （PTEN）/phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） pathway. MethodSixty SPF-grade male C57BL/6 mice were randomly divided into six groups： Normal group， model group， low， medium， and high dose groups of modified Guizhi Fulingwan （13， 26， 52 g·kg^-1·d^-1）， and positive control aspirin group （0.015 g·kg^-1·d^-1）. A mouse model of CRA was chemically induced using azoxymethane （AOM） and dextran sulfate sodium （DSS）. During the modeling process， mice received modified Guizhi Fulingwan or aspirin. Body weight of mice was measured weekly during the treatment. After 9 weeks， the number of adenomas formed was observed. Serum levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β） were determined by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining was used to observe the histopathologic changes in adenoma tissues. The expression of Cyclin D₁ and proliferative nuclear antigen （Ki67） was detected by immunohistochemistry （IHC）. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） was used to assess the apoptosis in adenoma tissues. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to observe the mRNA and protein expression levels of PTEN， PI3K， Akt， phosphorylated PI3K （p-PI3K）， phosphorylated Akt （p-Akt）， B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， cytochrome C （Cyt C）， Caspase-9， and caspase-3. ResultCompared with the normal group， the model group showed no significant change in body weight from week 1 to week 2， but a significant decrease from week 3 to week 9 （P<0.05，P<0.01）. The colorectal length was significantly shortened， and the colorectal weight increased with visible varying sized tumor-like protrusions on the mucosal surface （P<0.01）. Serum levels of TNF-α， IL-6， and IL-1β were elevated （P<0.01）. Histopathology showed disordered epithelial gland structure， elongated nuclei with pathological mitosis， and numerous lymphocytic infiltrations in the lamina propria. The positive expression rates of Cyclin D₁ and Ki67 were significantly increased （P<0.01）， while the apoptosis rate of adenoma cells was significantly decreased （P<0.01）. Expression levels of PI3K， Akt， Bcl-2 mRNA and proteins， as well as p-PI3K and p-Akt proteins， were significantly increased （P<0.01）， whereas PTEN， Bax， Cyt C， Caspase-9， and Caspase-3 mRNA and protein levels were significantly decreased （P<0.05， P<0.01）. Compared with the model group， all drug treatment groups showed an increase in body weight （P<0.01）， decreased intestinal weight， increased colorectal length， reduced number of adenomas significantly （P<0.05， P<0.01）， and significantly lowered serum levels of TNF-α， IL-6， and IL-1β （P<0.01）. Histopathology indicated improved glandular structure and reduced neutrophil infiltration in the mucosal lamina propria. The positive expression rates of Cyclin D₁ and Ki67 significantly decreased （P<0.01）， while the apoptosis rate of adenoma cells significantly increased （P<0.05， P<0.01）. Expression levels of PI3K， Akt， Bcl-2 mRNA and proteins， and p-PI3K and p-Akt proteins were significantly reduced （P<0.05， P<0.01）， while PTEN， Bax， Cyt C， Caspase-9， and Caspase-3 mRNA and protein levels significantly increased （P<0.05， P<0.01）. The high-dose modified Guizhi Fulingwan group exhibited the most significant intervention effects. ConclusionModified Guizhi Fulingwan may prevent CRA in mice by regulating the PTEN/PI3K/Akt signaling pathway and inducing the mitochondrial apoptosis pathway.

Objectives:To investigate the outcomes of cochlear implantation in Mandarin-speaking cochlear implant (CI) users with single-sided deafness (SSD).Methods:This study was a single-center prospective cohort study. Eleven Mandarin-speaking adult SSD patients who underwent CI implantation at Capital Medical University Beijing Tongren Hospital from August 2020 to October 2021 were recruited, including 6 males and 5 females, with the age ranging from 24 to 50 years old. In a sound field with 7 loudspeakers distributed at 180°, we measured root-mean-square error(RMSE)in SSD patients at the preoperative, 1-month, 3-month, 6-month, and 12-month after switch-on to assess the improvement of sound source localization. The Mandarin Speech Perception (MSP) was used in the sound field to test the speech reception threshold (SRT) of SSD patients under different signal-to-noise locations in a steady-state noise under conditions of CI off and CI on, to reflect the head shadow effect(S SSDN NH), binaural summation effect(S 0N 0) and squelch effect(S 0N SSD). The Tinnitus Handicap Inventory (THI) and the Visual Analogue Scale (VAS) were used to assess changes in tinnitus severity and tinnitus loudness in SSD patients at each time point. The Speech, Spatial and Qualities of Hearing Scale(SSQ) and the Nijmegen Cochlear Implantation Scale (NCIQ) were used to assess the subjective benefits of spatial speech perception and quality of life in SSD patients after cochlear implantation. SPSS 19.0 software was used for statistical analysis. Results:SSD patients showed a significant improvement in the poorer ear in hearing thresholds with CI-on compared with CI-off; The ability to localize the sound source was significantly improved, with statistically significant differences in RMSE at each follow-up time compared with the preoperative period ( P<0.05). In the S SSDN NH condition, which reflects the head shadow effect, the SRT in binaural hearing was significantly improved by 6.5 dB compared with unaided condition, and the difference was statistically significant ( t=6.25, P=0.001). However, there was no significant improvement in SRT between the binaural hearing condition and unaided conditions in the S 0N 0 and S 0N SSD conditions ( P>0.05). The total score of THI and three dimensions were significant decreased ( P<0.05). Tinnitus VAS scores were significantly lower in binaural hearing compared to the unaided condition ( P<0.001). The total score of SSQ, and the scores of speech and spatial dimensions were significant improved in binaural hearing compared to the unaided condition ( P<0.001). There was no statistical difference in NCIQ questionnaire scores between preoperative and postoperative ( P>0.05), and only the self-efficacy subscore showed a significant increase( Z=-2.497, P=0.013). Conclusion:CI could help Mandarin-speaking SSD patients restore binaural hearing to some extent, improve sound localization and speech recognition in noise. In addition, CI in SSD patients could suppress tinnitus, reduce the loudness of tinnitus, and improve subjective perceptions of spatial hearing and quality of life.

This study searched the clinical researches on traditional Chinese medicine （TCM） for cardiovascular diseases registered in Chinese Clinical Trial Registry and the US Clinical Trial Registry， the cardiovascular disease-related studies funded by the National Natural Science Foundation of China， as well as those published in China National Knowledge Infrastructure （CNKI）， Wanfang database， VIP.com， China Biology Medicine disc （CBMdisc）， Embase， Medline， Cochrane Library， and other databases published cardiovascular disease-related studies from 1 January 2021 to 30 June 2023. In order to analyse and evaluate the research progress of TCM treatment for coronary heart disease， hypertension， heart failure， and arrhythmia， this study aimed at recent research hotspots and research direction. It is found that the research on TCM for cardiovascular diseases was gradually deepening and the high-quality evidence continued to emerge. It is believed that studies related to the prevention and treatment of common cardiovascular diseases by TCM reflected the multi-angle integration of modern technology and pattern differentiation and treatment， closer integration of clinical and basic research， and further optimisation of pattern identification and interventions. On this basis， the research programme and implementation process should be further standardized， and the translation of research results should be emphasized to promote the standardized application and promotion of TCM diagnosis and treatment of cardiovascular diseases.

Non-muscle myosin heavy chain 9-related disease (MYH9-RD) is an autosomal dominant disease caused by the mutations of the MYH9 gene encoding the non-muscle mysoin heavy chain ⅡA and leads to abnormal accumulation of myosin in cells. These further causes functional disorders of the blood, eye, ear, kidney, and liver systems. MYH9-RD displays heterogeneous kidney involvement and outcomes, but doctors still lack understandings of the mechanism and treatment strategies, owing to difficulty of conducting renal biopsies. Here, we report a case of MYH9-RD with tail fragments heterozygous mutation, which renal pathology is presented as glomerular minor lesion. Moreover, we reviewed related relevant to strengthen clinical diagnosis and understanding of MYH9-RD.