Different subspecies or strains of the same species produce varied clinical manifestations.The clarifica-tion of parasite taxonomy is useful for the researches of their biology, epidemiology and control.DNA molecular markers have the advantages of high polymorphism, non-pleiotropy, and clear identifying alleles, etc.This paper summarizes the first generation(restriction fragment length polymorphism, random amplified polymorphic DNA), second generation(sim-ple sequence repeat-anchored PCR, inter-simple sequence repeat) and third generation(single nucleotide polymorphism) molecular marker techniques, and their application in taxonomic identification of parasites.

Objective:To observe the impacts of different postures on the hemodynamics of lower extremity vein.Methods:In this single center non-randomized controlled study in 15 healthy female volunteers, the hemodynamic changes in the common femoral vein were detected by color Doppler ultrasound at 10 different postures: supine position, slope positions with bed end elevated at 15°, 30° and 45°, trapezoidal positions with bed end elevated at 15°, 30° and 45°, and positions with bed head elevated at 30°, 45° and 60°.Results:Different postures resulted in significant differences in the velocity of blood flow in the common femoral vein ( P<0.05), with slope position at 45°> slope position at 30°> slope position at 15° = trapezoidal position at 30°> trapezoidal position at 15° = trapezoidal position at 45°> supine position> position with bed head elevated at 30°> position with bed head elevated at 45° = position with bed head elevated at 60°. Conclusions:In the postures observed in this study, the slope position with bed end elevated at 45° can promote the most effectively the blood reflux in the lower extremity vein, the trapezoidal positions with bed end elevated may not facilitate the distal blood reflux in the lower extremity vein, and positions with bed head elevated may hinder the blood reflux in the lower extremity vein.

Objective:To analyze the prevention and treatment of Kaschin-Beck disease in Fu County, Shaanxi Province, so as to provide basis for consolidating the results of Kaschin-Beck disease prevention and control and the treatment of patients with Kaschin-Beck disease.Methods:The epidemiological investigation of data and clinical data Kaschin-Beck disease from 1954 to 2022 were collected from the Fu County Institute for Endemic Disease Prevention and Control and People's Hospital of Fu County in Shaanxi Province, respectively. The retrospective study was used to investigate the prevention and control of Kaschin-Beck disease in Fu County through the adoption of comprehensive measures such as relocation, grain exchange, water improvement, conversion of farmland to forest and so on.Results:In 1954, there were 6 endemic townships and 127 endemic villages of Kaschin-Beck disease in Fu County, with a total population of 78 781. A total of 16 327 patients with Kaschin-Beck disease were detected by X-ray examination, with a detection rate of 20.72%, including 5 434 patients without clinical symptoms. There were 5 850 patients with clinical grade Ⅰ, 3 725 patients with clinical grade Ⅱ and 1 318 patients with clinical grade Ⅲ. In 1975, the first general survey of Kaschin-Beck disease was conducted in the whole county, with a total population of 101 341. A total of 9 575 patients with Kaschin-Beck disease were detected by X-ray examination, with a detection rate of 9.45%, including 3 247 cases without clinical symptoms. Among the patients with symptoms, there were 3 704 cases of clinical grade Ⅰ, 2 006 cases of clinical grade Ⅱ and 618 cases of clinical grade Ⅲ. The patients with Kaschin-Beck disease were mainly local residents, accounting for 93.92% (8 993/9 575). In 1997, 27 320 students aged 7 to 16 years in 342 schools of 15 townships were examined and found that there were 169 cases without clinical symptoms with X-ray changes, only 2 cases with clinical grade Ⅰ, and the detection rate decreased to 0.63%. In 1997, Fu County began to encourage relocation and grain exchange to prevent Kaschin-Beck disease. The relocation targets were mainly the seriously ill villages with new cases among children and poor living conditions. By 1999, a total of 100 households and 469 people were relocated, and by 2020, 1 569 households and 5 334 people were relocated. In 1997 and 1998, measures were taken to improve water quality in 43 natural villages in 6 townships, with 1 591 households and 7 375 people benefiting. In 2010, a general survey of Kaschin-Beck disease was conducted in 15 townships of Fu County, with a total population of 135 858, and 3 424 patients with Kaschin-Beck disease were detected by X-ray examination, with a detection rate of 2.52%. There were 2 885 cases with clinical symptoms, including 1 584 cases with clinical grade Ⅰ, 1 024 cases with clinical grade Ⅱ and 277 cases with clinical grade Ⅲ. Among them, 32 561 minors under 16 years old were examined, and 49 patients with Kaschin-Beck disease were detected by X-ray examination, with a detection rate of 0.15%, and all of them were clinical gradeⅠ patients. In 2014, a total of 73 600 people were surveyed in 170 endemic villages of Fu county, and 2 885 patients with Kaschin-Beck disease were detected by X-ray examination, with a detection rate of 3.92%. Compared with 2010, the number of patients with Kaschin-Beck disease increased zero, and there were no underage patients under the age of 16 for 4 consecutive years. By the end of 2016, 666.67 hm 2 of farmland had been converted to forest in Fu County, involving 9 townships (including communities), 33 administrative villages and 1 993 households. In 2018, another general survey of Kaschin-Beck disease was conducted in Fu County, with a total population of 157 362. A total of 2 308 patients were detected by X-ray examination, with a detection rate of 1.47%. Among them, there were 1 270 cases of clinical grade Ⅰ, 870 cases of clinical grade Ⅱ and 168 cases of clinical grade Ⅲ, and there were no patients with Kaschin-Beck disease under 16 years old in the county for 8 consecutive years (2011-2018). In the same year, 22 cases of Kaschin-Beck disease joint replacement were completed in Fu County, and by January 2022, about 60 cases had completed joint replacement. Conclusion:The prevention and control of Kaschin-Beck disease in Fu County has achieved remarkable results through comprehensive measures such as relocation, grain exchange, water improvement and conversion of farmland to forest.

Objective To investigate the mechanism of action of Dingchuan granules in intervening respiratory syncytial virus pneumonia based on network pharmacology and animal experiments.Methods The potential active ingredients and targets of each traditional Chinese medicine in Dingchuan granules were obtained from TCMSP and TCMID databases,and the active ingredients of the drugs in Dingchuan granules were screened according to ADME pharmacokinetic parameters;the potential disease targets of respiratory syncytial virus pneumonia were obtained from Genecards,OMIM,and DisGeNet databases;the protein-interaction networks of intersecting targets were visualized by using String platform;the key core targets were visualized by using David database;and the intersection targets were visualized by using David database.Interaction networks were constructed using the String platform to visualize the intersecting targets;GO functional enrichment and KEGG pathway enrichment analyses of the key core targets were performed using the David database;and then the relevant networks for the intervention of Respiratory Syncytial Virus Pneumonia in the Dingchuan particles were constructed using the Cytoscape software(3.9.1).Respiratory syncytial virus(RSV)-induced respiratory syncytial virus pneumonia in young rats was selected for experimental validation.Results The results of the network pharmacology showed that the 177 potential active ingredients of Dingchuan granules acted on 144 targets,and there were 1075 targets related to respiratory syncytial virus pneumonia,and 55 drug-disease intersecting targets were obtained,of which 25 core targets were ALB,IL6,CASP3,EGFR,VEGFA,etc.The GO function of Dingchuan granules was also investigated,and the GO function of Dingchuan was investigated.The results of GO function enrichment analysis showed that the biological processes involved mainly include positive regulation of transcription,positive regulation of RNA polymerase II promoter transcription,positive regulation of gene expression,negative regulation of apoptosis,etc.The KEGG pathway enrichment mainly involves the PI3K-Akt signaling pathway,the cancer pathway,the tumor necrosis factor signaling pathway,the IL-17 signaling pathway and so on.The KEGG pathway enrichment mainly involves PI3K-Akt signaling pathway,tumor necrosis factor signaling pathway,IL-17 signaling pathway,etc.Animal experiments initially showed that the fixed wheezing granules can play a role in inflammation and immune response by regulating the PI3K-Akt signaling pathway,thus participating in the inflammatory and immune response.Compared with the normal group,the ratios of p-PI3K/PI3K and p-Akt/Akt in the lung tissues of young rats in the model group were significantly higher(P<0.05),the viral load was significantly higher(P<0.05),the pathological score of lung tissue damage was significantly higher(P<0.05),and the content of inflammatory factors IL-6 and TNF-α in alveolar lavage fluid(BALF)was significantly higher(P<0.05).Compared with the model group,the expression of p-PI3K/PI3K and p-Akt/Akt proteins in the lung tissues of young rats in each dose group and the positive control group was reduced(P<0.05),the viral load was significantly reduced(P<0.05),the pathological scores of lung tissue injury were significantly reduced(P<0.05),and the contents of inflammatory factors IL-6 and TNF-α in BALF were significantly reduced(P<0.05).Conclusion The study revealed the synergistic mechanism of multi-component,multi-target,and multi-pathway action of Dingchuan granules for the treatment of respiratory syncytial virus pneumonia.It was verified by animal experiments that RSV infection in young rats probably activated the PI3K-Akt signaling pathway,which caused the release of inflammatory factors IL-6 and TNF-α.Dingchuan granules could effectively down-regulate the PI3K-Akt signaling pathway,inhibit the release of inflammatory factors IL-6 and TNF-α,and thus achieve the anti-inflammatory effect.

Objective To propose a frequency-domain pulse wave prediction model based on fusion attention mechanism,improving the low classification accuracy and poor generalization performance of hypertension time-domain pulse wave classification based on artificial intelligence technology.Methods Firstly,the time-domain pulse wave was transformed into frequency-domain Mel-scale frequency cepstral coefficient features to enhance its discriminability.Then,temporal convolutional network and Transformer structures were employed to extract the deep features of pulse waves,and self-attention mechanism and selective kernel attention were combined for decision fusion to extract relevant features.Floodings regularization method was used to indirectly control the training loss and prevent overfitting.A 5-fold cross-verification experiment was conducted based on 527 clinical pulse diagnosis data provided by Longhua Hospital,Shanghai University of Traditional Chinese Medicine and Shanghai Traditional Chinese Medicine-Integrated Hospital.Additionally,the extreme gradient boosting algorithm was employed to calculate the contribution rate ranking of frequency-domain pulse wave features,and the key factors affecting the classification accuracy of the model were analyzed to provide reference for the clinical auxiliary diagnosis of traditional Chinese medicine.Results The evaluation metrics accuracy,F1 score,precision,recall rate and area under curve value of the model proposed in this study were 0.939 6,0.924 9,0.940 9,0.929 5,and 0.993 4,respectively.The static characteristics of the pulse wave,the contribution rate of the first-order difference and the second-order difference coefficients were relatively balanced,indicating that the degree of hypertension risk was not only related to the static characteristics of the pulse wave,but also to the dynamic characteristics of the pulse wave.Conclusion The proposed model has higher classification accuracy and generalization performance compared to typical pulse wave classification models.

Objective:To investigate the epidemiological characteristics of lower extremity deep vein thrombosis (DVT) in patients with femoral fracture.Methods:Retrospectively analyzed were the data of 2,571 patients with femoral fracture who had been treated at the Third Hospital of Hebei Medical University from January 2019 to December 2019. There were 1,079 males and 1,492 females, aged from 14 to 96 years (average, 67.1 years). There were 1,158 femoral neck fractures, 951 femoral intertrochanteric fractures, 309 femoral shaft fractures, and 153 femoral condylar fractures. 2,414 patients were treated surgically while 157 patients non-surgically. Color Doppler ultrasonography of both lower extremities was performed to determine the occurrence of DVT before operation and every week after operation for patients undergoing surgical treatment, and within 48 hours after admission and every week during hospitalization for those undergoing non-surgical treatment. The incidence and location of DVT were recorded for different femoral fractures.Results:The incidence of DVT in this cohort was 35.5%(913/2,517), that of proximal DVT 5.3%(135/2,571), and that of distal DVT 30.3% (778/2,571). In patients with femoral neck fracture, femoral intertrochanteric fracture, femoral shaft fracture and femoral condylar fracture, the incidence of DVT was respectively 28.8% (334/1,158), 44.7% (425/951), 30.7% (95/309) and 38.6% (59/153), the incidence of proximal DVT was respectively 2.7% (31/1,158), 5.6%(53/951), 9.7% (30/309) and 13.7% (21/153), and the incidence of distal DVT was respectively 26.2% (303/1,158), 39.1% (372/951), 21.0% (65/309) and 24.8%(38/153). The incidence of DVT in the femoral vein and above, popliteal vein, tibiofibular vein and intermuscular vein in this cohort was respectively 2.3%(60/2,571), 2.9%(75/2,571), 6.4%(165/2,571) and 23.8%(613/2,571).Conclusions:The incidence of DVT may be high in patients with femoral fracture, and the proximal DVT with a high risk of pulmonary embolism may occur more in patients with femoral condylar fracture.

Objective:To analyze the disease spectrum among children who were using hydroxychloroquine (HCQ), and evaluate the drug′s safety and compliance.Methods:From January 2008 to December 2019, children from Children′s Hospital of Fudan University who used HCQ were selected as subjects, the disease spectrum of HCQ was analyzed, and the drug safety and compliance were evaluated for the patients who were followed up for more than 6 months. Demographic information, diagnosis, initial dose, time of continuous use, cumulative dosage and related adverse reactions report, project and the results of eye test were collected.Results:A total of 528 cases used HCQ during the 12 years, with 156 male cases and 372 female cases, and age at initial medication was (10.5±3.2) years. Among them, 514 cases (97.3%) had rheumatic disease, 5 had pulmonary interstitial lesions and 9 had other system diseases. The top three of the rheumatic diseases were systemic lupus erythematosus (SLE) in 316 cases (316/514,61.5%), juvenile idiopathic arthritis in 69 cases (69/514,13.4%), and juvenile dermatomyositis in 56 cases (56/514,10.9%). During the same period, 397 cases were diagnosed with SLE, and the utilization rate was 79.6% (316/397), which was the highest compared with other diseases and increased year by year. Pulmonary interstitial lesions included 4 cases with SFTPC gene defect related interstitial lung disease. Of the 528 ceses who were treated with HCQ, 397 cases were included for evaluating HCQ′s safety and compliance, the initial dose was (4.2±1.0) mg/kg, duration was 29.6 (14.9, 48.8) months, the longest usage time was 127 months, the largest cumulative dosage was 566.8 g. The continuous usage duration ( Z=-3.191, P=0.001) of SLE was significantly higher than those of other diseases, as well as cumulative dosage ( Z=-5.355, P=0.001). All cases received comprehensive eye exams before medication, 354 cases (354/397, 89.2%) were followed up in the ophthalmological department, and 65.5% (232/354) of them could be reviewed regularly at least 1 time per year. One case suffered from severe skin adverse reactions when the drug was used for 32.7 months, and no other serious adverse reactions were reported. HCQ related retinopathy was not seen during the follow-up period. There were 5 cases stopped HCQ on their own. Conclusions:HCQ was widely used in rheumatic disease in children, especially in those with SLE. It was safe for long-time usage in children, and the medication compliance and the ophthalmic follow-up was good.

A 31-year-old man sought medical evaluation for a 2-year history of edema and proteinuria, with prior pathology suggesting atypical membranous nephropathy (MN). Despite treatment with a combination of steroids, calcineurin inhibitors, and four courses of rituximab (1 g, intravenous injection), the patient′s nephrotic syndrome showed no relief (24 h urine protein peaked at 31.18 g/d), indicating refractory nephrotic syndrome. Later in the disease course, a sudden surge of creatinine level (322.5 μmol/L) prompted a renal biopsy, which revealed concurrent acute interstitial nephritis. Further treatment involving steroids, cyclophosphamide, and a fifth rituximab infusion (1 g, intravenous injection) resulted in improvement in renal function (serum creatinine: 322.5?147 μmol/L), but the MN failed to achieve partial relief. Subsequent treatment with the novel humanized CD20 monoclonal antibody obinutuzumab (1 g, intravenous injection) was initiated. In the latest follow-up, anti-phospholipase-A2-receptor antibody (PLA2R) antibody were negative, B cells were eliminated, serum albumin was 36 g/L, urine protein-to-creatinine ratio was 4 810 mg/g, and serum creatinine was 162 μmol/L. This case underscores the potential efficacy of obinutuzumab in refractory MN. For advanced MN cases, prompt identification of the cause of acute kidney injury is crucial, emphasizing the need for targeted interventions to potentially stall renal function decline.

OBJECTIVE To prepare and evalu ate doxorubicin-loaded red blood cell membrane chitosan-targeted nanoparticles of targeting tumor cell folate acid （FA）receptor（FA-RBC-DOX-CS-NPs）. METHODS Doxorubicin-loaded chitosan nanoparticles （DOX-CS-NPs） were prepared by ion cross-linking method. FA and amino polyethylene glycol phospholithin （NH2- PEG2000-DSPE）were covalently linked to modify the red blood cell membrane to construct FA-RBC-DOX-CS-NPS. FA-RBC- DOX-CS-NPs were characterized and investigated on in vitro drug release characteristics ，antitumor activity and endocytosis ability （investigation with human breast cancer MCF- 7 cells）. RESULTS Average particle size of FA-RBC-DOX-CS-NPs was （254.200± 2.651）nm，and polydispersity index was 0.199±0.031；Zeta potential was （－10.100±0.213）mV. FA-RBC-DOX-CS-NPs released fast in the tumor microenvironment （pH6.5）. Cellular experiments showed that ，the nanoparticles could inhibit the activity of MCF- 7 cell proliferation and improve the efficiency of endocytosis. CONCLUSIONS FA-RBC-DOX-CS-NPs are prepared successfully. The nanoparticles have good tumor cell targeting and endocytosis ability ，and can realize the enrichment of drugs in tumor cells.