OBJECTIVE: To explore the clinical application value of metagenomic next-generation sequencing (mNGS) in pathogen detection of bloodstream infection secondary to hematologic diseases in children. METHODS: 42 children with bloodstream infections secondary to hematologic diseases admitted to the Children's Hematology and Tumor Center of the Affiliated Hospital of Guangdong Medical University from November 2021 to May 2023 were included in the study, and their clinical data, results of peripheral blood mNGS and traditional blood culture, pathogen distribution characteristics, and diagnostic efficacy of mNGS were retrospectively analyzed. RESULTS: Among the 42 children included, there were 2 cases (4.8%) of aplastic anemia (AA), 27 cases (64.3%) of acute lymphoblastic leukemia (ALL), 7 cases (16.7%) of acute myeloid leukemia (AML), 1 case (2.4%) of chronic myeloid leukemia (CML), 2 cases (4.8%) of hemophagocytic lymphohistiocytosis (HLH), 2 cases (4.8%) of non-Hodgkin lymphoma (NHL), and 1 case (2.4%) of Wiskott-Aldrich syndrome (WAS). In mNGS testing, pathogens were detected in 31 peripheral blood samples, with a positive rate of 73.8% (31/42), significantly higher than the pathogen positive rate of 16.7% (7/42) detected by traditional blood culture, and the difference was statistically significant (P < 0.05). Among the pathogen-positive cases detected by mNGS, 23 cases (74.2%) were positive for bacteria, 12 cases (38.7%) were positive for viruses, and 9 cases (29.0%) were positive for fungi. 32.2% (10/31) of the pathogen-positive samples detected by mNGS were mixed pathogens, which could not be effectively detected by traditional blood culture. CONCLUSION Peripheral blood mNGS has advantages in the detection of pathogens of bloodstream infection secondary to hematologic diseases, with a higher detection rate of pathogen positivity than traditional blood cultures. It can detect viruses, rare pathogens and mixed pathogens, and has good clinical application value.
Child ; Child, Preschool ; Female ; Humans ; Male ; Hematologic Diseases/immunology* ; High-Throughput Nucleotide Sequencing ; Metagenomics ; Retrospective Studies ; Sepsis/microbiology*

Objective To investigate the incidence and the types of gene mutations of α-thalassemia in the child-bearing pop-ulation of Conghua District,Guangzhou.Methods Blood samples from 24 083 people of childbearing age were screened by blood cell analysis and hemoglobin electrophoresis,α-globin gene variation was detected by GAP-PCR and PCR reverse dot blot in the positive cases,and 17 common β-globin gene mutations were detected by PCR reverse Dot blot.Results A total of 2 596 cases of α-thalassemia gene abnormality were detected by gene identification,and the abnormal rate was 10.78%.A sum of 170 cases(0.71%)had a compound mutation of α-β gene.There were 2 550 cases(98.23%)of deletion and 46 cases(1.77%)of non-deletion in the mutant genes.There were 14 types of gene mutation,including 5 types of HbH disease(with--SEA/-α3.7 primarily),4 mild types(with 68.61%of--SEA/αα genotype),and 5 quiescent types(the top two genotypes were-α3.7/αα and-α4.2/αα).A total of 23 types of αβ complex gene mutation were detected,and the top six types were--SEA/βCD41-42,-α3.7/βCD41-42,--SEA/β654,--SEA/-28,-α3.7/β654 and-α3.7/βCD17,which accounted for 75.27%of all the complex types.Conclusion The gene abnormality rate of α-thalassemia in Conghua District of Guangzhou City was high.The gene mutation type and constitu-ent ratio,which have their own characteristics,is a special region of α-thalassemia.

Objective To investigate the influences of Lycium barbarum polysaccharide on apoptosis of corneal epithelial cells induced by hypertonicity through regulating adenosine monophosphate activated protein kinase(AMPK)/uncoordinated 51-like kinase 1(ULK1)autophagy pathway.Methods The ophthalmoxerosis cell model was established by osmotic pressure of 500 mOsm·L-1 on corneal epithelial cells.They were divided into model group,positive control group(0.3％sodium hyaluronate eye drops),inhibitor group(1 mg·mL-1 Lycium barbarum polysaccharide+10 μmol·L-1compound C),experimental-L,-H groups(0.5,1.0 mg·mL 1 Lycium barbarum polysaccharide),and normal cultured CRL-11135 cells were taken as blank group(no treatment was performed).Cell apoptosis was detected by flow cytometry.Autophagy was detected by MDC staining.Western blot was used to detect the expressions of p-AMPK/AMPK and p-ULK1/ULK1.Results The apoptosis rates of experimental-L,experimental-H,positive control,inhibitor,model and blank groups were(26.47±2.13)％,(13.68±2.21)％,(12.54±2.16)％,(18.73±2.12)％,(37.56±3.25)％and(6.35±2.14)％;the relative contents of autophagosomes were(59.63±8.14)％,(89.89±9.04)％,(90.31±9.13)％,(62.75±7.26)％,(43.11±6.45)％and(100.00±0.00)％;p-AMPK/AMPK were 0.45±0.07,0.64±0.08,0.66±0.06,0.53±0.04,0.34±0.05 and 0.87±0.06;p-ULKl/ULKl were 0.54±0.06,0.75±0.05,0.77±0.03,0.65±0.04,0.46±0.04 and 0.92±0.08,respectively.The above indexes in experimental-L,-H groups and positive control group were significantly different from those in model group(all P＜0.05);the above indexes in inhibitor group were significantly different from those in experimental-H group(all P＜0.05).Conclusion Lycium barbarum polysaccharide can inhibit the apoptosis of corneal epithelial cells induced by hypertonicity by activating the AMPK/ULK1 autophagy pathway.

Objective To investigate the clinical efficacy of endoscopic pterygoid nerve combined with anterior ethmoidal nerve transection in treatment of allergic rhinitis with nasal polyps.Method A clinical practice study was conducted on 108 patients with allergic rhinitis complicated with nasal polyps.After enrollment,they were randomly divided into two groups,54 cases were included in the control group and treated with conventional endoscopic sinus surgery,while 54 cases were included in the observation group and treated with conventional endoscopic sinus surgery combined with pterygoid nerve and ethmoidal nerve transection.The nasal pain visual analogue scale(VAS),rhinoconjunctivitis quality of life questionnaire(RQLQ),asthma quality of life questionnaire(AQLQ),total effective rate,and complication rate of the two groups were compared before treatment and 1 week,2 weeks,3 weeks,3 months,6 months,and 1 year after treatment.Result Compared with the control group,the total effective rate of treatment was 98.15%in the observation group,which was higher than 85.19%in the control group,the difference was statistically significant(P<0.05).Before treatment,there were no significant differences in nasal resistance(NR),mucociliary transport rate(MTR),mucociliary transport time(MTT),and Lund-Kennedy scores between the two groups,the differences were no statistically significant(P>0.05).After treatment,MTR in two groups increased compared with before treatment,and the observation group was higher than the control group,the difference was statistically significant(P<0.05);NR,MTT,and Lund-Kennedy scores in two groups decreased,and the observation group was lower than the control group,the differences were statistically significant(P<0.05).Before treatment,there were no significant differences in nasal pain VAS,RQLQ,and AQLQ between the two groups,the differences were no statistically significant(P>0.05).After treatment,at 1 week,2 weeks,3 weeks,and 3 months,6 months,and 1 year,the observation group had lower nasal pain VAS compared to the control group,while the observation group had lower RQLQ and higher AQLQ compared to the control group,the differences were statistically significant(P<0.05).The incidence of complications in the observation group was lower than the control group,the difference was statistically significant(P<0.05).Conclusion The clinical efficacy of endoscopic pterygoid nerve combined with ethmoidal nerve transection for the treatment of allergic rhinitis complicated with nasal polyps is significant.It can not only effectively alleviate nasal pain in patients,but also improve their quality of life and reduce postoperative complications,which has clinical application value.

OBJECTIVE: Physical exercise, a common non-drug intervention, is an important strategy in cancer treatment, including hepatocellular carcinoma (HCC). However, the mechanism remains largely unknown. Due to the importance of hypoxia and cancer stemness in the development of HCC, the present study investigated whether the anti-HCC effect of physical exercise is related to its suppression on hypoxia and cancer stemness. METHODS: A physical exercise intervention of swimming (30 min/d, 5 d/week, for 4 weeks) was administered to BALB/c nude mice bearing subcutaneous human HCC tumor. The anti-HCC effect of swimming was assessed in vivo by tumor weight monitoring, hematoxylin and eosin (HE) staining, and immunohistochemistry (IHC) detection of proliferating cell nuclear antigen (PCNA) and Ki67. The expression of stemness transcription factors, including Nanog homeobox (NANOG), octamer-binding transcription factor 4 (OCT-4), v-Myc avian myelocytomatosis viral oncogene homolog (C-MYC) and hypoxia-inducible factor-1α (HIF-1α), was detected using real-time reverse transcription polymerase chain reaction. A hypoxia probe was used to explore the intratumoral hypoxia status. Western blot was used to detect the expression of HIF-1α and proteins related to protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β)/β-catenin signaling pathway. The IHC analysis of platelet endothelial cell adhesion molecule-1 (CD31), and the immunofluorescence co-location of CD31 and desmin were used to analyze tumor blood perfusion. SMMC-7721 cells were treated with nude mice serum. The inhibition effect on cancer stemness in vitro was detected using suspension sphere experiments and the expression of stemness transcription factors. The hypoxia status was inferred by measuring the protein and mRNA levels of HIF-1α. Further, the expression of proteins related to Akt/GSK-3β/β-catenin signaling pathway was detected. RESULTS: Swimming significantly reduced the body weight and tumor weight in nude mice bearing HCC tumor. HE staining and IHC results showed a lower necrotic area ratio as well as fewer PCNA or Ki67 positive cells in mice receiving the swimming intervention. Swimming potently alleviated the intratumoral hypoxia, attenuated the cancer stemness, and inhibited the Akt/GSK-3β/β-catenin signaling pathway. Additionally, the desmin⁺/CD31⁺ ratio, rather than the number of CD31⁺ vessels, was significantly increased in swimming-treated mice. In vitro experiments showed that treating cells with the serum from the swimming intervention mice significantly reduced the formation of SMMC-7721 cell suspension sphere, as well as the mRNA expression level of stemness transcription factors. Consistent with the in vivo results, HIF-1α and Akt/GSK-3β/β-catenin signaling pathway were also inhibited in cells treated with serum from swimming group. CONCLUSION Swimming alleviated hypoxia and attenuated cancer stemness in HCC, through suppression of the Akt/GSK-3β/β-catenin signaling pathway. The alleviation of intratumoral hypoxia was related to the increase in blood perfusion in the tumor. Please cite this article as: Xiao CL, Zhong ZP, Lü C, Guo BJ, Chen JJ, Zhao T, Yin ZF, Li B. Physical exercise suppresses hepatocellular carcinoma progression by alleviating hypoxia and attenuating cancer stemness through the Akt/GSK-3β/β-catenin pathway. J Integr Med. 2023; 21(2): 184-193.
Humans ; Animals ; Mice ; Carcinoma, Hepatocellular/drug therapy* ; Proto-Oncogene Proteins c-akt/metabolism* ; Proliferating Cell Nuclear Antigen/therapeutic use* ; Mice, Nude ; Glycogen Synthase Kinase 3 beta/genetics* ; beta Catenin/therapeutic use* ; Liver Neoplasms/drug therapy* ; Desmin/therapeutic use* ; Ki-67 Antigen ; Cell Line, Tumor ; Hypoxia ; RNA, Messenger/therapeutic use* ; Cell Proliferation

OBJECTIVES: This work aimed to investigate the molecular mechanism of cyclic tensile stress (CTS) stimulating autophagy in human periodontal ligament cells (hPDLCs). METHODS: hPDLCs were isolated and cultured from normal periodontal tissues. hPDLCs were loaded with tensile stress by force four-point bending extender to simulate the autophagy of hPDLCs induced by orthodontic force du-ring orthodontic tooth movement. XMU-MP-1 was used to inhibit the Hippo signaling pathway to explore the role of the Hippo-YAP signaling pathway in activating hPDLC autophagy by tensile stress. The expression levels of autophagy-related genes (Beclin-1, LC3, and p62) in hPDLCs were detected by real-time quantitative polymerase chain reaction. Western blot was used to detect the expression levels of autophagy-related proteins (Beclin-1, LC3-Ⅱ/LC3-Ⅰ, and p62) and Hippo-YAP pathway proteins (active-YAP and p-YAP) in hPDLCs. Immunofluorescence was used to locate autophagy-related proteins (LC3-Ⅱand p62) and Hippo-YAP pathway proteins (active-YAP) of hPDLCs. RESULTS: CTS-activated autophagy in hPDLCs and expression of autophagy-related proteins initially increased and then decreased; it began to increase at 30 min, peaked at 3 h, and decreased (P<0.05). CTS increased the expression of active-YAP protein and decreased the expression of p-YAP protein (P<0.05). When XMU-MP-1 inhibited the Hippo-YAP signaling pathway (P<0.05), active-YAP protein was promoted to enter the nucleus and autophagy expression was enhanced (P<0.05). CONCLUSIONS The Hippo-YAP signaling pathway is involved in the regulation of autophagy activation in hPDLCs under CTS.
Humans ; Hippo Signaling Pathway ; Periodontal Ligament/metabolism* ; Beclin-1/metabolism* ; Cells, Cultured ; Autophagy

OBJECTIVES: To establish a method for the detection of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS). METHODS: The blood samples were treated with 1-butyl-3-methylimidazolium hexafluorophosphate as an extraction solvent. The samples were extracted by ultrasound-assisted extraction and separated by ZORBAX Eclipse Plus C18, 95Å column. The mobile phase A aqueous solution containing 0.1% formic acid and 10 mmol/L ammonium acetate, and mobile phase B mixed organic solvent containing acetonitrile/methanol (V_acetonitrile∶V_methanol=2∶3) were used for gradient elution at the flow rate of 1.00 mL/min. An electrospray ion source in positive mode was used for detection in the multiple reaction monitoring. RESULTS: The linearities of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples were good within the corresponding range, with correlation coefficients (r) greater than 0.995 6. The limits of detection were 3.00, 0.40 and 1.30 ng/mL, respectively. The limit of quantitation were 8.00, 1.00 and 5.00 ng/mL, respectively. The extraction recoveries ranged from 76.00% to 106.44%. The relative standard deviations of the intra-day and inter-day precisions were less than 16%. Carbamazepine and its main metabolite 10,11-dihydro-10,11-epoxycarbamazepine were detected in blood samples of death cases with a mass concentration of 2.71 μg/mL and 252.14 ng/mL, respectively. CONCLUSIONS This method has high sensitivity and good selectivity, which is suitable for the detection of carbamazepine and its metabolites in blood samples, and can be used for carbamazepine-related forensic identifications.
Chromatography, Liquid/methods* ; Tandem Mass Spectrometry ; Methanol ; Carbamazepine/analysis* ; Benzodiazepines/analysis* ; Solvents ; Chromatography, High Pressure Liquid ; Solid Phase Extraction

Objective:To explore the effectiveness of applet-assisted teaching in the pedagogy of stomatology and gain suggestions for the improvement of the applet.Methods:Using digital technology, we built a professional atlas applet for Oral Histopathology that covered a wide range of contents including illustrations, histological slides, schematic diagrams, and hand-drawn pictures of knowledge points. Based on the WeChat platform, the applet had the functions of bilingual (Chinese/English) annotation and real-time interactive communication between teachers and students. Thirty dental students were given quizzes before and after using the atlas applet. The scores were analyzed using the t-test with SPSS 22.0 software. At the same time, 45 students filled a questionnaire after using the applet for one month, which involved comments on the use of the digital atlas, the degree of satisfaction and suggestions for the applet during learning, and suggestions for its co-use by teachers and students in the future. Results:After using the atlas applet to assist with learning, the test scores of the students were improved significantly in all domains. There were significant differences between the test and control groups in the score of completion questions [(17.00±2.61) vs. (15.03±1.85), P<0.05], the score of identification of images [(27.93±5.08) vs. (25.13±3.31), P<0.05], and the total score [(78.77±8.59) vs. (72.90±6.08), P<0.05]. The questionnaire feedback showed that the students were highly satisfied with the atlas applet-assisted learning, the function of teacher-student interaction, and the overall performance of the applet. Conclusion:An atlas is one of the crucial teaching tools of Oral Histopathology for its intuitionistic and realistic presentation of tissue structures, which plays an important role in the theoretical learning process for dental students. The combination of the atlas and electronic media based on portable mobile devices is beneficial for students' understanding of knowledge.

Objective To observe the clinical curative effect of unilateral biportal endoscopy(UBE)in the treatment of extreme lateral lumbar disc herniation(ELLDH).Methods From June 2019 to June 2022,25 patients with ELLDH were treated with UBE-guided discectomy,including 16 males and 9 females.The age ranged from 26 to 62 years with a mean of(53.67±17.45)years.History ranged from 3 d to 10 years.There were 9 cases of internal foraminal type(type Ⅰ),13 cases of external foraminal type(type Ⅱ)and 3 case of mixed type(type Ⅲ).There were 8 cases of L4/5 space and 17 cases of L5/S1 space.All the patients underwent anterior and lateral lumbar X-rays,CT and MRI scans before surgery.The visual analogue scale(VAS)pain score and Oswestry disability index(ODI)assessed lower limb and lower back pain and functional recovery before surgery and at 3 d and 3 months after surgery,respectively.Macnab criteria evaluated the immediate surgical outcome.Results The UBE technique was used to treat ELLDH.The operative time was(79.79±23.97)min,the intraoperative bleeding volume was 40～80 mL,with an average of(55.80±10.74)mL.Follow-up time was(7.02±4.26)months.Preoperative VAS of lower limb was(7.04±0.92),lower back VAS was(3.49±1.52),ODI was(35.03±2.97)%.Compared with the preoperative results,the lower limb VAS was(2.17±0.61),lower back VAS was(1.48±0.43),and ODI was(18.77±3.15)%on day 3 after surgery,lower limb VAS was(1.38±0.65),lower back VAS was(1.03±0.48)and ODI was(6.05±1.80)%on the 3 months after surgery were improved(P＜0.05).The excellent and good rate was 96.0%(excellent 20,good 4,fair 1),and no obvious complications were observed during the follow-up.Conclusion UBE is effective,with little trauma,clear intraoperative visual field,good intraoperative experience,and less traumatic and safe in the treatment of ELLDH.

On February 2022, WHO released the evidence-based guideline on control and elimination of human schistosomiasis, with aims to guide the elimination of schistosomiasis as a public health problem in disease-endemic countries by 2030 and promote the interruption of schistosomiasis transmission across the world. Based on the One Health concept, six evidence-based recommendations were proposed in this guideline. This article aims to analyze the feasibility of key aspects of this guideline in Chinese national schistosomiasis control program and illustrate the significance to guide the future actions for Chinese national schistosomiasis control program. Currently, the One Health concept has been embodied in the Chinese national schistosomiasis control program. Based on this new WHO guideline, the following recommendations are proposed for the national schistosomiasis control program of China: (1) improving the systematic framework building, facilitating the agreement of the cross-sectoral consensus, and building a high-level leadership group; (2) optimizing the current human and livestock treatments in the national schistosomiasis control program of China; (3) developing highly sensitive and specific diagnostics and the framework for verifying elimination of schistosomiasis; (4) accelerating the progress towards elimination of schistosomiasis and other parasitic diseases through integrating the national control programs for other parasitic diseases.
China/epidemiology* ; Disease Eradication ; Humans ; Public Health ; Schistosomiasis/prevention & control* ; World Health Organization