ObjectiveA multiplex amplification system was constructed based on the capillary electrophoresis platform for simultaneous detection of saliva, semen, and vaginal secretions using tissue-specific RNA markers. The aim of this study is to identify the tissue origin of suspicious body fluid stains found at crime scenes and determine whether the body fluid stains at the crime scene are one or several types among saliva, semen, and vaginal secretions. MethodsThirty saliva samples, forty semen samples, and forty vaginal secretion samples (half from 2015 and half from 2024) were collected from healthy adult volunteers. Through primer designing, system formulation, and PCR condition optimization, a multiplex fluorescent amplification system was constructed. The specificity, sensitivity, and detection ability for mixed samples of this system were investigated, and it was tested using real crime scene materials. In the primer design stage, to reduce the requirements for RNA template quality, the amplification products were set within 80-300 bp. In the system formulation stage, dominant and subordinate primers were mainly considered. By reducing the concentration of dominant primers and increasing that of subordinate primers, a capillary electrophoresis spectrum with an appropriate peak height ratio was finally obtained. Additionally, gradient experiments were designed to adjust the concentrations of PCR reagents and PCR amplification conditions, and multiple versions of DNA amplification enzymes were optimized to achieve the best experimental results. ResultsThrough statistical analysis, there was no significant difference in the capillary electrophoresis of the 3 types of body fluid samples from the two years (2015 and 2024), demonstrating that the sample preservation method in this study can preserve samples for a relatively long time. The composite amplification system constructed in this study exhibited high specificity for all 3 types of body fluid, with no cross-reactions between the markers of each type of body fluid. The minimum detection thresholds for the 3 types of body fluid reached 0.002 9, 0.001 5, and 0.42 mg/L, respectively. This system also had a high degree of discrimination for mixed samples, especially for semen-saliva mixtures, where each body fluid marker could still be successfully detected when the concentration ratio of semen to saliva was 100:1. Meanwhile, in the two actual cases presented in this article, the application of this composite amplification system performed outstandingly. ConclusionThe composite amplification detection system constructed in this study can achieve the correct screening of saliva, semen, and vaginal secretions, overcoming the problems such as low specificity and sensitivity of marker tests and unbalanced RFU values of each marker in previous studies. The specificity and sensitivity meet the practical work requirements, and the operation is simple. It provides an analytical and identification method for body fluid stains in actual case and is applicable to the identification of the tissue origin of biological evidence at crime scenes involving sexual assault, indecent assault, and other criminal acts. In the future, more types of body fluid markers will be screened to expand the types of body fluids detected by the system, and body fluid-specific cSNP and cInDel genetic markers will be introduced to infer the sources (individuals and types) of mixed and complex stains more accurately.

Based on clinical epidemiological data， it is believed that qi deficiency constitution is closely related to allergic diseases. According to the fundamental principles of traditional Chinese medicine （TCM） constitution theory， the intrinsic connection between qi deficiency constitution and allergic diseases is analyzed from the perspectives of inherited endowment， life process， environmental restriction， and the interplay of form and spirit. This paper discusses the key points of regulating qi deficiency constitution to prevent allergic diseases in three stages， prevention before illness， prevention of disease progression， and prevention of recurrence after recovery. It also distinguishes the treatment directions for regulating qi deficiency constitution to treat allergic diseases based on different disease locations such as the lung， spleen， and kidney. This aims to expand new ideas for the research on qi deficiency constitution and allergic diseases as well as the prevention and treatment of allergic diseases.

Cerebral ischemia-reperfusion injury （CIRI） has a very high incidence， disability， and mortality rates， which seriously affects human life and health. In recent years， modern medicine has made some progress in the diagnosis and treatment of CIRI， but there are still problems such as difficulties in postoperative rehabilitation and adverse drug reactions， and new therapeutic drugs for CIRI are urgently needed. As an important class of active ingredients in traditional Chinese medicine， flavonoids can play antioxidant， apoptosis inhibition， anti-inflammatory， and other pharmacological effects to improve brain tissue damage， which is important for improving the quality of life of CIRI patients and slowing down the aging of the social population. Numerous studies have found that flavonoids in traditional Chinese medicine can regulate cell surface receptors Toll-like receptor 4/nuclear factor-kappaB （TLR4/NF-κB）， phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， adenylate-activated protein kinase/mammalian target of rapamycin protein （AMPK/mTOR）， Ras homologous gene family member A/Rho-associated coiled-coil protein kinase （RhoA/ROCK）， nuclear factor E₂-associated factor 2/Kelch-like epoxychloropropane-associated protein-1/haemoglobin oxygenase 1 （Nrf2/Keap1/ HO-1）， Notch， and other signaling pathways， so as to regulate the transcription and expression of related proteins after CIRI， alleviate brain tissue injury， and improve CIRI. This paper analyzed the relevant literature in China and abroad in recent years， reviewed the mechanism of action and related pathways of flavonoids in traditional Chinese medicine to improve CIRI， and explored the new therapeutic direction of CIRI at the metabolic level， with a view to providing a basis for the further development and application of flavonoids in traditional Chinese medicine.

Objective:To investigate the clinical efficacy of arthroscopic double-row double-pulley technique in the treatment of Ideberg type IA scapular glenoid fracture.Methods:A retrospective case series study was conducted to analyze the clinical data of 16 patients with Ideberg type IA scapular glenoid fracture admitted to Jiading Branch of Shanghai General Hospital from January 2018 to December 2021, including 10 males and 6 females, aged 25-65 years [(42.9±5.1)years]. The patients were treated with arthroscope-assisted reduction and double-row double-pulley technique. The operation time was recorded. Three-dimensional reconstruction of the shoulder joint with CT was performed to assess fracture displacement and healing. Modified University of California Los Angeles (UCLA) score and Constant-Murley score were used to evaluate shoulder function and Visual Analogue Scale (VAS) score was used to evaluate pain before surgery, at 3, 6, 12 months after surgery and at the last follow-up. The complications were observed.Results:All the patients were followed up for 12-36 months [(20.3±4.4)months]. The operation time was 60-90 minutes [(74.7±8.9)minutes]. Three-dimensional construction of the shoulder joint with CT performed at 3 months after surgery showed that there was no fracture re-displacement and all the patients had bone union. The modified UCLA score, Constant-Murley score and VAS score at 3 months after surgery were (30.4±0.4)points, (84.3±1.4)points and 2.0(1.3, 3.0)points, respectively, which were significantly improved compared with those before surgery [(21.1±0.5)points, (56.4±1.3)points and 5.0(5.0, 6.0)points respectively] ( P<0.05). The modified UCLA score, Constant-Murley score and VAS score at 6 months after surgery were (33.1±0.4)points, (91.0±0.5)points and 1.0(1.0, 2.0)]points respectively, which were significantly improved compared with those at 3 months after surgery ( P<0.05). The modified UCLA score, Constant-Murley score and VAS score at 12 months after surgery were (33.5±0.3)points, (92.6±0.6)points and 1.0(0.3, 1.8)points respectively, showing no significant differences from those at 6 months after surgery ( P>0.05). The modified UCLA score, Constant-Murley score and VAS score at the last follow-up were (33.8±0.8)points, (93.7±1.8)points and 1.0(0.0, 1.0)points respectively, with no significant differences from those at 12 months after surgery ( P>0.05). There were no complications such as wound infection, neurovascular injury or shoulder stiffness after surgery. Conclusion:Arthroscopic double-row double-pulley technique for the treatment of Ideberg type IA scapular glenoid fracture has a short operation time, a high fracture healing rate, good shoulder function recovery, and pain relief, with no common complications.

Objective:To investigate the current status of endoscopic treatment for gastroesophageal varices in portal hypertension in China, and to provide supporting data and reference for the development of endoscopic treatment.Methods:In this study, initiated by the Liver Health Consortium in China (CHESS), a questionnaire was designed and distributed online to investigate the basic condition of endoscopic treatment for gastroesophageal varices in portal hypertension in 2022 in China. Questions included annual number and indication of endoscopic procedures, adherence to guideline for preventing esophagogastric variceal bleeding (EGVB), management and timing of emergent EGVB, management of gastric and isolated varices, and improvement of endoscopic treatment. Proportions of hospitals concerning therapeutic choices to all participant hospitals were calculated. Guideline adherence between secondary and tertiary hospitals were compared by using Chi-square test.Results:A total of 836 hospitals from 31 provinces (anotomous regions and municipalities) participated in the survey. According to the survey, the control of acute EGVB (49.3%, 412/836) and the prevention of recurrent bleeding (38.3%, 320/836) were major indications of endoscopic treatment. For primary [non-selective β-blocker (NSBB) or endoscopic therapies] and secondary prophylaxis (NSBB and endoscopic therapies) of EGVB, adherence to domestic guideline was 72.5% (606/836) and 39.2% (328/836), respectively. There were significant differences in the adherence between secondary and tertiary hospitals in primary prophylaxis of EGVB [71.0% (495/697) VS 79.9% (111/139), χ2=4.11, P=0.033] and secondary prophylaxis of EGVB [41.6% (290/697) VS 27.3% (38/139), χ2=9.31, P=0.002]. A total of 78.2% (654/836) hospitals preferred endoscopic therapies treating acute EGVB, and endoscopic therapy was more likely to be the first choice for treating acute EGVB in tertiary hospitals (82.6%, 576/697) than secondary hospitals [56.1% (78/139), χ2=46.33, P<0.001]. The optimal timing was usually within 12 hours (48.5%, 317/654) and 12-24 hours (36.9%, 241/654) after the bleeding. Regarding the management of gastroesophageal varices type 2 and isolated gastric varices type 1, most hospitals used cyanoacrylate injection in combination with sclerotherapy [48.2% (403/836) and 29.9% (250/836), respectively], but substantial proportions of hospitals preferred clip-assisted therapies [12.4% (104/836) and 26.4% (221/836), respectively]. Improving the skills of endoscopic doctors (84.2%, 704/836), and enhancing the precision of pre-procedure evaluation and quality of multidisciplinary team (78.9%, 660/836) were considered urgent needs in the development of endoscopic treatment. Conclusion:A variety of endoscopic treatments for gastroesophageal varices in portal hypertension are implemented nationwide. Participant hospitals are active to perform emergent endoscopy for acute EGVB, but are inadequate in following recommendations regarding primary and secondary prophylaxis of EGVB. Moreover, the selection of endoscopic procedures for gastric varices differs greatly among hospitals.

Stroke， including ischemic stroke and hemorrhagic stroke， has the characteristics of high morbidity， high disability rate， high mortality rate， high recurrence rate， and high economic burden. Ischemic stroke is the most common type of stroke， which is mainly caused by intracranial artery occlusion. The clinical manifestations of patients are hemiplegia， aphasia， sensory disturbance， and other neurological deficits， accompanied by gastrointestinal symptoms such as constipation and gastrointestinal bleeding. Intestinal flora plays an important role in the pathogenesis of ischemic stroke， and its potential biological effects have received extensive attention. Intestinal flora can not only affect intestinal barrier function but also regulate gastrointestinal immunity and affect host homeostasis. In recent years， traditional Chinese medicine （TCM） has shown remarkable effects and small adverse reactions in the prevention and treatment of cerebral ischemia. The research on the effect of TCM in improving cerebral ischemia injury by regulating the structure and metabolism of intestinal flora and maintaining the function of intestinal flora has gradually become a hot topic. Based on the interaction between TCM and intestinal flora in relevant literature in recent years， this review investigated the mechanism of anti-cerebral ischemic injury of TCM via regulating intestinal flora structure， affecting intestinal flora metabolism， and regulating body immunity and made a summary， proving a basis for further elucidating the role of intestinal flora in cerebral ischemia and the mechanism of TCM in prevention and treatment of ischemic stroke.

Objective To explore the effects of polystyrene microplastics(PS-MPs)on the growth,activity,oxida-tive stress levels,biofilm formation and virulence of Klebsiella pneumoniae.Methods Klebsiella pneumoniae was exposed to PS-MPs at different concentrations(10,50 and 100 μg/ml)and particle sizes(0.1,1.0 and 5.0 μm),and the growth curves were measured.The bacterial activity was determined by CCK-8(cell counting kit-8).The level of intracellular reactive oxygen species(ROS)was determined by fluorescence probe.The biofilm forming ability was determined by crystal violet staining.Real-time quantitative fluorescent PCR(qRT-PCR)was used to detect the relative expression levels of biofilm-forming genes(luxS,mrkA,wbbM,pgaA,wzm)and viru-lence genes(ureA,uge,wabG,fimH).Results A high concentration(100 μg/ml)of 0.1 μm PS-MPs had a stronger inhibitory effect on the growth and activity of Klebsiella pneumoniae,and the intracellular ROS level signifi-cantly increased,indicating that smaller particle size and higher concentration of PS-MPs were more toxic to bacte-ria.PS-MPs of 100 μg/ml particle size groups(0.1,1.0 and 5.0 μm)significantly promoted the biofilm forma-tion of Klebsiella pneumoniae.The relative expression levels of biofilm formation related genes(luxS,mrkA,wbbM,pgaA,wzm)and virulence genes(ureA,uge,wabG,fimH)increased.Conclusion By inducing Kleb-siella pneumoniae to produce a high level of ROS,PS-MPs can cause oxidative stress,inhibit the growth and activi-ty of bacteria,and enhance the biofilm formation ability and virulence,thus affecting the biological characteristics of Klebsiae pneumoniae.

A 28-year-old woman was found to have coagulation factor Ⅷ activity (FⅧ∶C) <1% and von Willebrand factor antigen (VWF∶Ag) <1% during routine prenatal examinations. No pathogenic variation was found in the exon region of the VWF gene using next-generation sequencing. The clinical presentation of this patient does not match the clinical characteristics of type Ⅲ hemophilia [von Willebrand disease (VWD) ]; therefore, third-generation sequencing technology was used to perform whole-genome sequencing on the patient and her family members. Multiple members of the patient’s paternal family carried a heterozygous variant of VPS33B, c.869G>C. The family members carrying this variant all had varying degrees of reduced VWF levels (39% -56% ). Moreover, the proband was detected with the heterozygous variant c.1474dupA in GP1BA. The ACMG and Clinvar databases determined that this variation was associated with platelet-type pseudo VWD. The decrease in VWF levels caused by heterozygous variations in VPS33B in families is the first international report, and no previous studies have reported cases of severe decrease in plasma VWF levels caused by double heterozygous variations in VPS33B and GP1BA.

Background::Although overnight fasting is recommended prior to endoscopic retrograde cholangiopancreatography (ERCP), the benefits and safety of high-carbohydrate fluid diet (CFD) intake 2 h before ERCP remain unclear. This study aimed to analyze whether high-CFD intake 2 h before ERCP can be safe and accelerate patients’ recovery.Methods::This prospective, multicenter, randomized controlled trial involved 15 tertiary ERCP centers. A total of 1330 patients were randomized into CFD group ( n = 665) and fasting group ( n = 665). The CFD group received 400 mL of maltodextrin orally 2 h before ERCP, while the control group abstained from food/water overnight (>6 h) before ERCP. All ERCP procedures were performed using deep sedation with intravenous propofol. The investigators were blinded but not the patients. The primary outcomes included postoperative fatigue and abdominal pain score, and the secondary outcomes included complications and changes in metabolic indicators. The outcomes were analyzed according to a modified intention-to-treat principle. Results::The post-ERCP fatigue scores were significantly lower at 4 h (4.1 ± 2.6 vs. 4.8 ± 2.8, t = 4.23, P <0.001) and 20 h (2.4 ± 2.1 vs. 3.4 ± 2.4, t= 7.94, P <0.001) in the CFD group, with least-squares mean differences of 0.48 (95% confidence interval [CI]: 0.26–0.71, P <0.001) and 0.76 (95% CI: 0.57–0.95, P <0.001), respectively. The 4-h pain scores (2.1 ± 1.7 vs. 2.2 ± 1.7, t = 2.60, P = 0.009, with a least-squares mean difference of 0.21 [95% CI: 0.05–0.37]) and positive urine ketone levels (7.7% [39/509] vs. 15.4% [82/533], χ2 = 15.13, P <0.001) were lower in the CFD group. The CFD group had significantly less cholangitis (2.1% [13/634] vs. 4.0% [26/658], χ2 = 3.99, P = 0.046) but not pancreatitis (5.5% [35/634] vs. 6.5% [43/658], χ2 = 0.59, P = 0.444). Subgroup analysis revealed that CFD reduced the incidence of complications in patients with native papilla (odds ratio [OR]: 0.61, 95% CI: 0.39–0.95, P = 0.028) in the multivariable models. Conclusion::Ingesting 400 mL of CFD 2 h before ERCP is safe, with a reduction in post-ERCP fatigue, abdominal pain, and cholangitis during recovery.Trail Registration::ClinicalTrials.gov, No. NCT03075280.

Objective To analyze the correlation between triglyceride glucose index (TyG) combined with different obesity indexes and new-onset ischemic stroke (IS) in 10 years in prediabetes population in Luzhou,Sichuan through cohort study. Methods A total of 2551 residents who participated in the epidemiological survey in Luzhou in 2011 and were diagnosed as having prediabetes were enrolled. According to the quartiles of baseline TyG-waist circumference (WC),TyG-waist to height ratio (WtHR),TyG-body mass index (BMI),and TyG-waist to hip ratio (WHR),they were divided into the 1st,2nd,3rd,and 4th quartiles groups (Q1,Q2,Q3,and Q4 groups). The incidence of new-onset IS in each group in 10 years was analyzed. The relationships between TyG-WC,TyG-WtHR,TyG-WHR and new-onset IS were analyzed by multivariate logistic regression. Results Among the 2551 prediabetes subjects enrolled in the study,123 died of various causes during the 10 years of non-intervention follow-up,with a mortality of 4.8％;142 had new-onset IS,with an incidence of 5.6％;and 224 had new-onset diabetes mellitus,including 22 also had new-onset IS,accounting for 9.8％. The quartile grouping analyses of TyG-WC and TyG-WtHR showed that the incidence of new-onset IS in prediabetes subjects over 10 years was Q4 group>Q3 group>Q2 group>Q1 group,with significant differences (P<0.05). The quartile grouping analysis of TyG-BMI showed that there was no significant difference in the incidence of new-onset IS (P>0.05). The quartile grouping analysis of TyG-WHR showed that the incidence of new-onset IS was Q3 group>Q4 group>Q2 group>Q1 group,with significant difference (P<0.05). Multivariate logistic regression analysis showed that TyG-WtHR was associated with new-onset IS in prediabetes subjects,either without adjusting any variable or after adjusting multiple factors. Conclusion TyG-WtHR is a risk factor of IS in prediabetes individuals. Enhancing management of prediabetes patients with high TyG-WtHR may reduce the risk of IS.