ObjectiveTo observe the clinical effectiveness and safety of Dingchan Granule （定颤颗粒） for paroxysmal atrial fibrillation with syndrome of qi stagnation and blood stasis. MethodsUsing a randomised， double-blind， placebo controlled study method， 90 patients with paroxysmal atrial fibrillation with qi stagnation and blood stasis syndrome were divided into 45 cases each in the treatment group and the control group. Both groups were given conventional western medicine treatment， and the treatment group was additionally treated with Dingchan Granule， while the control group was treated with Dingchan Granule placebo， both of which were taken orally for 8 g each time twice a day. Both groups were treated for 8 weeks. We compared the clinical effectiveness， the improvement of traditional Chinese medicine （TCM） symptoms and the recovery rate of atrial fibrillation between the two groups. We compared the number and duration of atrial fibrillation episodes， TCM symptoms score， atrial fibrillation symptom classification， 24-hour average ventricular rate， Pittsburgh Sleep Quality Index （PSQI）， anxiety index， depression index before and after treatment， and evaluated the safety of the two groups. ResultsThe total clinical effectiveness rate in the treatment group was 82.22% （37/45）， which was better than 60.00% （27/45） in the control group （P＜0.05）. The total effective rate of TCM syndrome effectiveness in the treatment group was 88.89% （40/45）， which was better than 66.67% （30/45） in the control group （P＜0.05）； and the rate of atrial fibrillation regression in the treatment group was 26.67% （12/45）， better than 6.67% （3/45） in the control group （P＜0.05）. The number and duration of atrial fibrillation episodes in both groups were significantly decreased （P＜0.01）， and the number and duration of atrial fibrillation episodes in the treatment group were lower than those in the control group （P＜0.01）. The TCM syndrome scores of both groups after treatment were significantly lower than before treatment （P＜0.01）， and the scores of the treatment group was lower than those of the control group （P＜0.05）. The severity of atrial fibrillation symptoms and the grading of atrial fibrillation symptoms in both groups after treatment were improved （P＜0.01）， and the degree of symptom improvement in the treatment group was better than that in the control group （P＜0.01）. The 24-hour average ventricular rate of both groups after treatment was significantly lower （P＜0.01）. The PSQI， anxiety index and depression index of the treatment group were all lower than before treatment （P＜0.01）， while the PSQI and anxiety index of the control group were both lower than before treatment （P＜0.01 or P＜0.05）， the PSQI， anxiety index and depression index of the treatment group being lower than those of the control group （P＜0.05 or P＜0.01）. No adverse events occurred in both groups， and no abnormalities were observed in blood， urine， stool routine， liver and kidney function， and coagulation function indexes. ConclusionDingchan Granule for paroxysmal atrial fibrillation with qi stagnation and blood stasis syndrome can alleviate clinical symptom， improve TCM symptom scores， increase atrial fibrillation recovery rate， stabilise the average ventricular rate， and significantly improve the quality of sleep， alleviate the anxiety and depression， with a good safety profile.

It is believed that the pathogenesis of oculomotor nerve palsy is insufficient marrow sea （髓海）， withered yang qi， poor contraction of eyelids and periocular meridians， and inability to open and close the eyes. The eye system is connected to the marrow sea， as well as the the foot taiyang （太阳） channel， foot yangming （阳明） channel， foot jueyin （厥阴） channel， yinqiao mai （阴跷脉） and yangqiao mai （阳跷脉）， and is nourished by the liver， spleen and kidney. Treatment should take into account both the branch and the root cause. It is suggested to treat the root by regulating the marrow sea， and treat the branch by unblocking the meridians and dredging the collaterals， thereby balancing the mild and the urgency of the yinqiao mai and yangqiao mai. Using the "Gen （根）-Liu（溜）-Zhu （注）-Ru （入）"acupoints to bypass the various meridians and taking the gallbladder meridian according to twelve major meridians that run on both sides of the body， both of which can provide ideas for improving symptoms such as ptosis and limited eye movement caused by oculomotor nerve palsy.

Objective To screen differentially expressed long non-coding RNAs (lncRNAs) in the liver of mice infected with Schistosoma japonicum during the chronic pathogenic stage and identify their functions, so as to provide insights into unravelling the role of lncRNAs in S. japonicum infection-induced liver disorders. Methods Twenty 6-week-old C57BL/6 mice were randomly divided into two groups, of 10 animals each group. Each mouse in the experimental group was infected with (15 ± 2) S. japonicum cercariae via the abdomen for modeling chronic S. japonicum infection in mice, and distilled water served as controls. All mice were sacrificed 70 days post-infection, and mouse liver specimens were sampled for RNA extraction and library construction. All libraries were sequenced on the Illumina NovaSeq 6000 sequencing platform. Data cleaning was performed using the fastp software, and reference genome alignment and gene expression (FPKM) calculation were performed using the HISAT2 software. Potential lncRNA sequences were predicted using the software CNIC, CPC, Pfam, and PLEK, and potential lncRNAs were screened. Differentially expressed lncRNAs were screened with the DESeq2 software and subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to identify biological processes and metabolic pathways involved in target genes of differentially expressed lncRNAs. Results A total of 333 potential lncRNAs were screened, and 67 were identified as differentially expressed lncRNAs, including 49 up-regulated and 18 down-regulated lncRNAs. A total of 53 target genes were predicted for differentially expressed lncRNAs. GO enrichment analysis showed that these target genes were mainly enriched in biological process and molecular function, among which Sema7a, Arrb1, and Ccl21b genes may be hub target genes for positive regulation of extracellular regulated protein kinase 1 (ERK1) and ERK2 cascades and may participate in the regulation of collagen expression. KEGG enrichment analysis showed that the target genes of differentially expressed lncRNAs were mainly enriched in cytokine-cytokine receptor interaction, viral protein interactions with cytokines and cytokine receptors, chemokine signaling pathway, and nuclear factor kappa-B (NF-κB) signaling pathway. Conclusions This study identifies differentially expressed lncRNAs and functional enrichment of their target genes in the liver of mice during the chronic pathogenic stage of S. japonicum infection. Up-regulated lncRNAs may affect biological processes of ERK1/2 cascades and chemokine signaling pathways via target genes Sema7a, Arrb1, and Ccl21b, thereby affecting collagen expression and inflammatory signal pathways, ultimately affecting the development of liver disorders.

Objective To explore the dilution conditions and standards in detecting the semen samples with high sperm concentration using computer-aided sperm analysis(CASA)systems.Methods CASA systems with 10 μm depth disposable counting chambers were used for the examination.The samples were divided into undiluted group(Group 1∶sperm concentration＜50×106/mL)and diluted groups(Group 2∶50×106/mL≤ sperm concentration＜100× 106/mL;Group 3∶sperm concentration≥100× 106/mL).When sperm concentration＜50×106/mL,no dilution was performed.When sperm concentration≥50× 106/mL,the samples were diluted with saline at 1∶n/(50×106)ratio(n=sperm concentration,n/[50×106]rounded down)to＜50×106/mL of sperm concentration.The sperm concentration,progressive motility(PR),non-progressive motility(NP),total motility(PR+NP)and immotile sperm percent-age(IM)were analyzed before and after dilution.The consistency of results pre-and post-dilution was compared.ROC curve was used to analyze the optimal dilution threshold.Results The differences in the parameters pre-and post-dilution gradually rosed with the increased sperm concentration.ROC curve analysis showed optimal dilution thresholds were 133.05 × 106/mL,101.75 × 106/mL,118.60×106/mL,90.90×106/mL,111.83×106/mL for the sperm concentration,PR+NP,PR,NP and IM respectively.Considering sperm concentration and NP were most affected the undiluted high concentration samples,the optimal comprehensive dilution threshold was determined as 125.08× 106/mL.Conclusion When sperm concentration exceeds 125×106/mL,it is recommended to dilute semen sample with normal saline.

Objective To investigate the current status of semen testing in the hospitals without assisted reproductive technology in Hu-nan Province,propose improvement strategies to enhance the diagnosis of male infertility and promote reproductive health services in these hospitals.Methods Questionnaire surveys and on-site investigations were conducted to examine the semen testing status in 67 hospitals without assisted reproductive technology in Hunan Province.The existing problems were summarized,and improvement strate-gies were suggested.Results Among the 67 hospitals,59.7％(40/67)performed semen testing.Of these,45％(18/40)possessed dedicated personnel,60％(24/40)possessed staff received relevant training,and 22.5％(9/40)held certificates.Only 30％(12/40)conducted sperm morphology testing,among them 20％(8/40)able to stain and interpret following WHO standards.The hospitals of 37.5％(15/40)used computer-assisted sperm analysis(CASA)systems,and 22.5％(9/40)were equipped with phase-contrast microscopes.30％(12/40)had dedicated examination areas,and 32.5％(13/40)had independent ejaculation rooms,17.5％(7/40)used disposable slides to observe sperm concentration.In 2022,the hospitals of 57.5％(23/40)had an average daily sample volume＜1.The hospital of 67.5％(27/40)performed standard operating procedures,40％(16/40)followed the WHO 5th la-boratory manual for routine testing,15％(6/40)conducted internal quality control,and 12.5％(5/40)participated in external quali-ty assessment(EQA).Another 55％(22/40)wished to participate in EQA.Conclusion The semen testing capacity in the hospitals,in which assisted reproductive technology is not yet carried out currently,urgently requiring multifaceted improvements.The proposed strategies include emphasizing semen testing,establishing reproductive medicine consortia for comprehensive support,establishing a provincial quality control center for EQA,founding a professional committee for a learning platform,offering training to enhance staff expertise,and including the pre-pregnancy semen testing in public health programs.

Objective To investigate the effect of Yiqi Huoxue Tongluo Decoction on microRNA-126a-5p(miR-126a-5p)and vascular endothelial growth factor(VEGF)signaling pathway in cervical spondylotic myelopathy model rats.Methods Thirty healthy male SD rats were divided into the sham operation group,the model group and the traditional Chinese medicine(TCM)group by random number table method.Cervical spondylotic myelopathy models were prepared in the model group and the TCM group.The TCM group was given intragastric administration of Yiqi Huoxue Tongluo Decoction,while the sham operation group and the model group were given intragastric administration of normal saline for 12 weeks.After intervention,the threshold of mechanical stimulation and retraction time of thermal stimulation in each group were measured by behavior tests.Rats were sacrificed to collect intervertebral disc tissue for hematoxylin-eosin(HE)staining and observe the number of vascular buds in intervertebral disc.Rat intervertebral disc annulus fibrosus cells were subjected to terminal dexynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining.The miR-126a-5p and VEGF mRNA of rat intervertebral disc tissue were detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR).The expression of VEGF protein of rat intervertebral disc tissue was detected by Western blot assay.Results Compared with the sham operation group,the number of vascular buds in intervertebral disc was decreased in the model group and the TCM group.The cell destruction of intervertebral disc annulus was obvious in rats,and apoptosis was high and cell density decreased.Mechanical stimulation threshold decreased,and mechanical stimulation threshold decreased.The level of miR-126a-5p was decreased,and the expression levels of VEGF mRNA and protein were increased.Compared with the model group,the number of vascular buds in intervertebral disc was increased in the TCM group.The destruction of intervertebral disc annulus cells was alleviated in rats.The apoptosis of annulus fibrosus cells in intervertebral disc decreased and cell density increased.The threshold of mechanical stimulation increased,and the retraction time of thermal stimulation was prolonged.The level of miR-126a-5p increased,and the expression levels of VEGF mRNA and protein decreased(P＜0.05).Conclusion The mechanism of Yiqi Huoxue Tongluo Decoction in the treatment of cervical spondylotic myelopathy may be related to the up-regulation of miR-126a-5p expression and the down-regulation of VEGF expression.

BACKGROUND:In the process of exploring the mechanism of Alzheimer's disease,the important role of bioinformatics for common target screening has been revealed,enabling the use of its screening results as a basis for exploring the therapeutic effects of drugs on the disease. OBJECTIVE:To predict the targets of liraglutide,a glucagon-like peptide-1 receptor agonist,in the treatment of Alzheimer's disease by bioinformatics and molecular biology. METHODS:DisGeNET database and SEA database were used to obtain the common genes of Alzheimer's disease and liraglutide.GO/KEGG enrichment analysis of common targets was conducted using DAVID online database.Protein-protein interaction networks were constructed using STRING database.The optimal dosage of liraglutide was determined using cell counting kit-8 assay.Expression of key proteins was analyzed using immunofluorescence and immunoblotting techniques.The mouse hippocampal neuron HT22 cell line was used for ex vivo experiments,and the cells were randomly divided into three groups:HT22 group,HT22+Aβ group,and HT22+Aβ+Lir group.No special treatment was done in the HT22 group,while Aβ1-42 was used to intervene in the HT22 cell line for 24 hours to construct an Aβ injury cell model in the HT22+Aβ group.In additional to modeling,liraglutide was added to the HT22+Aβ+Lir group for 12 hours. RESULTS AND CONCLUSION:A total of 3 333 genes associated with Alzheimer's disease were screened from DisGeNET database.Then 147 potential targets of liraglutide were obtained from SEA database.Finally,64 common targets of Alzheimer's disease and Liraglutide were determined using R packets.GO/KEGG analysis of common targets using DAVID online database suggested that common targets were mainly enriched in the following biological processes:neuroactive ligand-receptor interaction,renin-angiotensin system,bladder cancer,endopeptidase activity,peptide receptor activity,G protein-coupled peptide receptor activity,and transport vesicles.The obtained 64 common target proteins were imported into SRTING online database for protein-protein interaction network construction,and the top three genes,matrix metalloproteinases 2,9 and interleukin 1β,were obtained.The activity of cultured cells was detected by the cell counting kit-8 kit.Liraglutide at 100 nmol/L was the optimal concentration for antagonizing Aβ1-42.In the western blot and immunofluorescence assays,the expression of matrix metalloproteinases 2,9 and interleukin 1β was significantly increased in the HT22+Aβ group compared with the HT22 group(P＜0.05)but significantly decreased in the HT22+Aβ+Lir group compared with the HT22+Aβ group(P＜0.05).To conclude,the above bioinformatics data and secondary validation of differential genes in the GEO database suggest that both matrix metalloproteinases 2,9 and interleukin 1β could be potential targets of liraglutide in the treatment of Alzheimer's disease.

Objective To construct an NZB/W(F1)mouse model of systemic lupus erythematosus and evaluate the effects of nicotinamide on each index of lupus nephritis pathogenesis of NZB/W(F1)mice in order to provide data for research on the role of nicotinamide in the treatment of lupus nephritis.Methods Female NZB/W(F1)mice were obtained by crossing male NZW mice with female NZB ones.Urine samples were collected using metabolic cages and proteinuria test strips were used to detect proteinuria.Blood samples were collected through the orbital venous plexus in mice.Enzyme-linked immunosorbent assay(ELISA)was used to detect the level of anti-dsDNA antibody.Levels of serum creatinine and urea nitrogen and liver function indexes were detected using an automatic blood analyzer.Hematoxylin-eosin staining and immunofluorescence technique were used to detect the pathological state of the kidney.Results The levels of proteinuria,double-stranded DNA antibodies,serum creatinine,and urea nitrogen were gradually increased during the natural course of the disease in female NZB/W(F1)mice,indicating that the lupus nephritis disease model was constructed in female NZB/W(F1)mice.Compared to the control group,nicotinamide feeding could obviously decrease the level of proteinuria(P=0.0070),inhibit the production of double-stranded DNA antibodies(P=0.0325),and retard the progression of serum creatinine(P=0.0067)and urea nitrogen indexes(P=0.0166)in serum.In addition,the pathological state of the kidney in the nicotinamide feeding group was significantly alleviated compared with the control group.Conclusion A lupus nephritis disease model is constructed in NZB/W(F1)mice.Nicotinamide feeding can obviously alleviate the disease state of lupus nephritis in NZB/W(F1)mice.

Objective To study the levels of serum Periostin and interleukin(I1-)-18 in preterm infants with bronchopulmonary dysplasia(BPD),and to analyze their correlation with the severity of the disease and their predictive value for BPD.Methods A total of 62 preterm infants with BPD diagnosed and treated in the hospital from January 2019 to January 2022 were retrospectively selected as the BPD group,and 80 preterm in-fants without BPD during the same period were selected as the non-BPD group.According to the severity of BPD,the infants with BPD were divided into mild subgroup(22 cases),moderate subgroup(24 cases)and se-vere subgroup(16 cases).The serum levels of Periostin and IL-18 were detected by enzyme-linked immu-nosorbent assay.Pearson correlation analysis was used to analyze the correlation between the clinical parameters.Multivariate Logistic regression was used to analyze the influencing factors of BPD,and receiver operating characteristic(ROC)curve was used to analyze the predictive value of each indicator for BPD.Results Compared with the non-BPD group,the BPD group had a significantly higher proportion of infants with pulmonary surfactant(PS)use,neonatal respiratory distress syndrome,apnea,patent ductus arteriosus,and serum levels of Periostin and IL-18,as well as a significantly longer duration of mechanical ventilation,noninvasive respiratory support,and length of hospital stay.The lung function parameters[tidal volume per kilogram(VT/kg),ratio of time to peak tidal expiratory flow to time(TPTEF/TE),ratio of volume to peak tidal expiratory flow to volume(VPEF/VE),expiratory flow at 50％tidal volume(50％TEF),expiratory flow at 75％tidal volume(75％TEF)]and 1,5 min Apgar score of BPD group were lower than that of non-BPD group,and the differences were statistically significant(P＜0.05).The serum levels of Periostin and IL-18 in mild subgroup,moderate subgroup and severe subgroup were increased in turn.The levels of serum Periostin and IL-18 were negatively correlated with pulmonary function indexes(VT/kg,50％TEF,75％TEF,TPTEF/TE,VPEF/VE,P＜0.05).Serum Periostin,IL-18 and neonatal respiratory distress syndrome were independent risk factors for BPD(P＜0.05),and PS was a protective factor(P＜0.05).Serum Periostin,IL-18 and neonatal respiratory distress syndrome were independent risk factors for the severity of BPD(P＜0.05).The area under the curve(95％CI)of serum Periostin and IL-18 alone and in combination for predicting BPD were 0.841(0.814-0.899),0.863(0.820-0.897),0.922(0.878-0.949),respectively.The sensitivity and specificity of combined prediction were 0.902 and 0.825,respectively.The area under the curve of the combination of the two indica-tors for predicting BPD was greater than that of each index alone,and the difference was statistically signifi-cant(Z=5.357,4.894,P＜0.001).Conclusion The levels of serum Periostin and IL-18 are increased in in-fants with BPD,which are related to the severity of BPD and lung function.The combination of serum Perios-tin and IL-18 has a high predictive value for BPD.

Currently, the application rate of virtual reality technology in the fields of medical education and medical treatment is relatively low. In this study, based on the structure of the human stomach, virtual reality technology, sensing technology, big data technology, and cloud computing technology were integrated. A new form of medical education was established to include the online website platform for data storage and analysis and the offline VR glasses for the physical operation. Through the use of 3d Max technology, Unity3D technology, and C# language, we constructed a three-dimensional model of the human stomach to present the stomach in three dimensions. This enables the users to immerse in it to achieve true human-computer interactive learning. This study updates the concept of medical education, effectively improves the quality and efficiency of medical education, and facilitates the development of surgical programs and reduction in the risk of surgery, as well as provides experimental materials for scientific research. In the future, it can be further developed to include the three-dimensional structures of the circulatory system and other major systems in the human body.