ObjectiveTo compare the contents and relative molecular weight distributions of proteins and polypeptides in Naoxuekang dropping pills， Huoxue Tongmai capsules and Maixuekang capsules of Hirudo single medicinal preparations， to evaluate the in vitro anticoagulant， antiplatelet and fibrinolytic activities of the three preparations， and to investigate the effects of temperature， pH and digestive enzymes on the anticoagulant activities of the three preparations. MethodsThe contents of soluble proteins and polypeptides in the three preparations were determined by bicinchoninic acid assay（BCA） and Bradford method， and the relative molecular weight distributions of the three preparations were determined by electrophoresis combined with gel chromatography. The antithrombin activity of the three preparations was evaluated by fibrinogen-thrombin time（Fibg-TT） method， and their anticoagulant activities were further assessed by the elongations of activated partial thromboplastin time（APTT）， prothrombin time（PT） and thrombin time（TT）. The antiplatelet aggregation activities of the three preparations were measured by turbidimetry and the fibrinolytic activities were measured by fibrin plate method. Relative TT was used as index to investigate the effects of temperature， pH and digestive enzyme buffer on anticoagulant activities of the three preparations. ResultsAt the lowest single dosage， the contents of proteins and polypeptides were in the order of Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. Both Huoxue Tongmai capsules and Maixuekang capsules had 11 electrophoretic bands between 4.0 kDa and 90 kDa， the bands of Maixuekang capsules were more clear in the range of >25 kDa， and there was 1 obvious band at 14 kDa for the two capsules. Huoxue Tongmai capsules had one specific band at 9.0 kDa and Maixuekang capsules had one specific band at 48.0 kDa. Naoxuekang dropping pills only had 2 electrophoretic bands at 6.5 kDa and 8.5 kDa， primarily containing peptides below 2 kDa， most of which were oligopeptides. The anticoagulant activity concentrations of the three preparations exhibited a certain dose-dependent effect. At the lowest single dosage， The anticoagulant activity concentrations were ranked as Naoxuekang dropping pills>Huoxue Tongmai capsules>Maixuekang capsules. The prolongation effect of the three preparations on coagulation time was dose-dependent. At the same concentration， the prolongation effect of Naoxuekang dropping pills and Huoxue Tongmai capsules was APTT prolongation rate>TT prolongation rate>PT prolongation rate， whereas for Maixuekang capsules， the sequence was TT prolongation rate>APTT prolongation rate>PT lengthening rate. At the single minimum dosage， the order of APTT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules≈Naoxuekang dropping pills， the order of PT prolongation rate was Naoxuekang dropping pills≈Maixuekang capsules>Huoxue Tongmai capsules， and the order of TT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. The three preparations showed dose-dependent effects on platelet aggregation induced by adenosine diphosphate（ADP） and arachidonic acid（AA）， and the effect induced by ADP was stronger than that induced by AA. The anti-platelet aggregation effect of Naoxuekang dropping pills was significantly stronger than that of Maixuekang capsules（P<0.01）， whereas Huoxue Tongmai capsules had the effect of promoting platelet aggregation. None of the three preparations had the ability to dissolve fibrin. The anticoagulant activity of Naoxuekang dropping pills was least affected by heating， while the activities of the two capsules decreased significantly within 5 min above 80 ℃， and continued to decrease within 2 h. Compared with pure water， the anticoagulant activities of the three preparations could be increased by 1-3 times under strong acidity（pH 1-3）. In the pepsin buffer， the anticoagulant activity of Naoxuekang dropping pills could be increased by 1-3 times， while the anticoagulant activities of Huoxue Tongmai capsules and Maxuekang capsules were significantly decreased， the lowest levels were about 60% and 20%， respectively. In trypsin buffer， the anticoagulant activities of Naoxuekang dropping pills， Huoxue Tongmai capsules and Maixuekang capsules decreased significantly， and the lowest levels decreased to about 41%， 41% and 35%， respectively. ConclusionThe contents of proteins and polypeptides and relative molecular weights of the preparations derived from lyophilized fresh Hirudo powder， dried Hirudo powder and reflux extract of Hirudo decrease sequentially， and the anticoagulant activity decrease gradually， but the anticoagulant pathway is different. And the anti-platelet aggregation activity of the reflux extract is significantly enhanced. The heat resistance and gastrointestinal stability of the three preparations increase successively， and the first two are suitable for enteric-soluble preparations， while the latter is suitable for routine oral administration. The above results can provide data reference for the rationality of different preparation methods， active substances， pharmacodynamics and mechanism of Hirudo preparations.

OBJECTIVE To characterize paclitaxel nanoparticles （PTX-PLGA-NPs） and evaluate their in vitro inhibitory effect on Lewis lung cancer cells. METHODS The PTX-PLGA-NPs prepared by the emulsion-solvent evaporation method were characterized in terms of particle size， polydispersity index （PDI）， Zeta potential， microscopic morphology， encapsulation efficiency， drug loading， ultraviolet-visible absorption characteristics and stability. Using mouse Lewis lung cancer cells as the subjects and paclitaxel reference substance as the control， the cytotoxicity and in vitro killing activity of PTX-PLGA-NPs were detected using CCK-8 method and Calcein-AM/PI double staining method， respectively. The effects of PTX-PLGA-NPs on cell apoptosis and cell cycle were assessed by Annexin Ⅴ-FITC/PI staining method and PI staining method， respectively. RESULTS PTX-PLGA-NPs were spherical with an average particle size of （172.03±0.95） nm， PDI of 0.098±0.012， and Zeta potential of （－1.76±0.02） mV. The encapsulation efficiency and drug loading were （52.32±0.66）% and （7.07±0.18）%， respectively， and the ultraviolet-visible absorption characteristics were not affected by the carrier polylactic-co-glycolic acid. When stored in the dark at 4 °C for 7 days， no significant change was noted in particle size， and the average PDI （after 1， 2， 4 and 7 days of storage） was under 0.3. Compared with the paclitaxel reference substance group， the PTX-PLGA-NPs group had more cells in a state of death， the survival rate （at the PTX concentration of 11.2 μg/mL） was significantly decreased， and both the apoptosis rate and the proportion of G2 phase cells were significantly increased （P＜0.05）. CONCLUSIONS The prepared PTX-PLGA-NPs indicate homogeneity in particle size， uniform dispersion， stable properties， and stronger in vitro killing effect on lung cancer cells than PTX.

Objective: To determine the main components of Astragalus membranaceus (Fisch.) Bge (A. membranaceus, Huang Qi), Astragaloside IV (AIV) and Astragalus polysaccharides (AP), to characterize their properties, evaluate their in vivo efficacy, and to analyze drug diffusion using dissolving microneedle (DMN) technology in vivo. Methods: Respectively, AIV- and AP-loaded DMNs comprising chitosan (CTS) and polyvinyl alcohol (PVA) were prepared via dual-mold forming. Their morphology, mechanical properties, in vivo solubility, and skin irritation characteristics were tested. In vivo efficacy was assessed in cyclophosphamide-induced immunosuppressed mice, in vivo diffusion of AIV and AP by DMNs and conventional methods was compared, and the rheological properties of AIV-CTS-PVA and AP-CTS-PVA mixtures were measured. Results: Subcutaneous dissolution and absorption of AIV-CTS-PVA and AP-CTS-PVA microneedles (MNs) at low doses (50%–17% of intraperitoneal AIV injection and 12%–4% of intravenous AP injection) reduced the spleen index and acid phosphatase activity in immunosuppressed mouse models, increased the thymus index, and achieved equivalent or better systemic therapeutic effects. Compared with injections, AIV and AP achieved controllable solid-liquid conversion through delivery with CTS-PVA MNs, resulting in highly localized aggregation within 48 h, reducing the initial explosive effect of the drug, and achieving stable and slow drug release. Conclusion The present study enhances our understanding of the efficacy and remote effects of drug-loaded DMNs from a traditional Chinese medicine (TCM) perspective, thereby promoting the development of precise and efficient delivery of TCM and further expanding the drug-loading range and application scenarios for DMNs.

Objective To investigate the risk factors associated with the presence of multiple Lugol-voiding lesions(LVLs)in patients with early esophageal cancer and the correlation with alcohol dehy-drogenase 1B(ADH1B)and aldehyde dehydrogenase 2(ALDH2)polymorphisms.Methods Patients who underwent endoscopic submucosal dissection due to early esophageal cancer were divided into group with multiple LVLs and group without multiple LVLs based on their endoscopic features.Their clinical data and the genotype of ADH1B and ALDH2 were collect-ed and SPSS 27.0 was used to statistically analyze the above data.Results A total of 83 subjects were included in the study,23 had multiple LVLs,most of them were seen in males,alcohol drinkers,and smokers with smoking index≥1 000,and multivariate analysis showed that alcohol consumption was an independent risk factor for it(OR=6.215,P=0.008).The gene polymorphism of ADH1B and ALDH2 and their interactions did not have any sig-nificant correlation with multiple LVLs.However,among alcohol drinkers,there was a 12-fold increased risk of multiple LVLs in patients carrying the ALDH2 A allele and drinking≥50 g per day compared to those carrying the ALDH2 GG genotype and drinking<50 g per day(P=0.045).Conclusion Alcohol consumption is an inde-pendent risk factor of multiple LVLs of the esophageal mucosa in patients with early esophageal cancer,and heavy alcohol consumption in carriers of the ALDH2 A allele will significantly increase the risk of multiple LVLs,and such patients should be closely followed up with endoscopy.

Background:Combined double-balloon enteroscopy(DBE)plays an important role in the diagnosis and treatment of patients with suspected small bowel bleeding(SSBB).Aims:To investigate the performance of combined DBE in patients with SSBB and their clinical features.Methods:A total of 94 patients with SSBB underwent combined DBE from June 2018 to April 2023 at the Third People's Hospital of Bengbu and the First Affiliated Hospital of Anhui Medical University were enrolled,and the clinical features were analyzed.Results:Fifty-four SSBB patients completed the combined DBE,and the combination rate was 57.4%.Ten patients(10.6%)stopped combined DBE due to discovery of bleeding lesions.Five patients(5.3%)stopped combined DBE due to intestinal stenosis.Twenty-five patients(26.5%)stopped combined DBE due to difficulty in insertion.Combined DBE rate in SSBB patients was correlated with the combination time and the number of previous DBE for endoscopists(P<0.05),but not the gender,age,bleeding manifestations,tobacco smoking and alcohol drinking,nutritional risk,abdominal operation history,perianal lesions,autoimmune diseases,preoperative anemia,preoperative albumin level,and enteroscopy approach(P>0.05).ROC curve showed that when the cut-off value of first insertion depth was 385 cm,the sensitivity and specificity for successful combined DBE in SSBB patients were 72.2%and 77.5%,respectively,and the area under ROC curve was 0.800(95%CI:0.705-0.875,P<0.001).Conclusions:The combined DBE rate in SSBB patients is correlated with the combination time and the number of previous DBE for endoscopists.80%of the SSBB patients are expected to complete the combined DBE when the first insertion depth is greater than 385 cm.

Objective:To evaluate the application of the standard of "Delimitation and Classification of Keshan Disease Areas" (GB 17020-2010, Standard for short), learn about the applicability of its technical indicators and requirements, and provide a basis for revision of the Standard.Methods:In March 2022, provinces with severe epidemic areas of Keshan disease or new cases reported in recent years were selected, including Gansu Province, Inner Mongolia Autonomous Region, Shandong Province, Shaanxi Province, Yunnan Province, Shanxi Province, Liaoning Province, and Chongqing City. Multi-stage stratified sampling method and questionnaire survey were adopted to collect information on the application of Standard by relevant health institutions.Results:A total of 448 questionnaires were collected, including 445 valid. The survey results showed that 87.64% (390/445) of the respondents were aware of the Standard, and 64.72% (288/445) had received training on the Standard. Eighty-two point two per cent (365/445) of the respondents believed that the Standard was simple and easy to operate, 83.82% (373/445) believed that the determination of the diseased townships was scientific and reasonable, and could be effectively implemented, and 83.60% (372/445) believed that the determination of historical epidemic areas was applicable to the current situation of Keshan disease; 38.88% (173/445), 38.20% (170/445), and 37.98% (169/445) of the respondents believed that the classification indicators for classifying epidemic areas (severe, moderate, and mild epidemic areas) were not applicable to the current situation of the disease in the local or provinces with Keshan disease, respectively. Among the indicators for epidemic areas classification, 30.79% (137/445), 29.21% (130/445), and 28.54% (127/445) of the respondents thought that the annual prevalence, the annual number of new cases and the annual incidence were most suitable for classification of Keshan disease areas.Conclusions:The Standard has been applied well in practice. However, based on the current situation of Keshan disease, it is suggested to redefine the standard for the severity of the disease and the classification of historical epidemic areas.

Objective To investigate the effects and mechanisms of median nerve electrical stimulation on synaptic plasticity in ischemic stroke rats.Methods 18 healthy male SD rats were randomly divided into Sham group(n = 6),ischemic stroke group(MCAO group,n = 6)and median nerve electrical stimulation group(MNES group,n = 6).The left middle cerebral artery occlusion model of rats was established by thread plug method.Thread plug was not inserted in sham group.The median nerve electrical stimulation group was given median nerve electrical stimulation intervention on the 3rd day after modeling,and intervention on the next day.After intervention for 7 times,behavioral detection,HE staining was used to detect median nerve injury.Nissl staining was used to detect cerebral infarction volume.Western blot was used for detection of the expression level of proteins related to synaptic plasticity,and electron microscopy was performed.Results HE staining showed that median nerve electrical stimulation did not cause damage to the median nerve in stroke rats,and the median nerve membrane was intact without obvious inflammatory cells.Compared with MCAO group,the neural function,motor function and coordination of the injured forelimb in MNES group were significantly improved(P<0.01).Compared with MCAO group,cerebral infarction volume in MNES group was significantly reduced(P<0.05),the pyknosis of Nissl bodies in ischemic penumbra decreased.Compared with MCAO group,the expression levels of synaptic plastication-related proteins PSD95 and synI in the cortex of MNES group were significantly up-regulated after median nerve electrical stimulation(P<0.05),the number of synapses in the ischemic cortex increased significantly(P<0.01).Conclusion Median nerve electrical stimulation is a safe and effective therapeutic measure to improve nerve function after stroke,and its mechanism is related to promoting synaptic plasticity.

Objective : To compare the diagnostic efficacy of three screening methods，including the epidemiological survey and the new gastric cancer scoring system proposed in the Expert Consensus Opinion on Early Gastric Cancer Screening Process in China ( Shanghai，2017) ，and the Kyoto classification score for screening gastric in early gastric cancer diagnosis． Methods: Patients who underwent endoscopic submucosal dissection and were pathologically confirmed were included in the gastric cancer group，patients with non-gastric cancer in the same period were consecutively included in the control group．Questionnaires and serological tests were performed in all enrolled patients，and the endoscopic presentations was recorded according to the classification of Kyoto gastritis during gastroscopy．The risk stratification and the detection rates of gastric cancer were estimated by different screening methods，and the receiver operating characteristic curve (ROC curve) of subjects diagnosed with each screening method alone and in combination were plotted to compare the area under the ROC curve (AUC) and the diagnostic efficacy . Results: Among all participants，84 were patients with gastric cancer and 120 were normal people without gastric cancer．The Kyoto risk score had the highest diagnostic value (AUC = 0. 79) among the others，and its sensitivity and specificity were 72. 6% and 79. 1% ，respectively，but the combined diagnosis of different methods did not significantly improve the diagnostic efficacy．The Kyoto risk score had a more discriminative value in the medium to high-risk group delineated by the new scoring system． Conclusion The Kyoto risk score has high clinical value in the diagnosis of early gastric cancer and can be used in conjunction with the new scoring system for early gastric cancer risk assessment.

Objective To observe the value of conventional ultrasound combined with ultrasonic elastography for evaluating the severity of active lesions of juvenile localized scleroderma(JLS).Methods Totally 27 JLS children with 46 active lesions were prospectively enrolled.The thickness,echo and blood flow of skin and subcutaneous tissue of JLS lesions and the paired sites were evaluated using conventional ultrasound,while the skin stiffness of all lesions and paired sites were assessed using shear wave elastography(SWE),and the strain ratio of lesions and surrounding skin were evaluated using strain elastography(SE).The thickness,echo and blood flow of skin as well as subcutaneous tissue,and the Young's modulus of skin were compared between JLS active lesions and the paired sites.The correlations of the individual sum total of each Young's modulus and the limited scleroderma activity index(LoSAI)or the limited scleroderma injury index(LoSDI)were explored.Results Compared with the paired site,no significant difference of the skin thickness was found among 46 JLS lesions(P＞0.05),but JLS lesions had thinner subcutaneous tissue(P＜0.05)and greater Young's moduli(all P＜0.05).The strain ratio of the lesions and surrounding skin were all larger than 2.For JLS children with active lesions,the individual sum total of Young's moduli had moderate rank correlations with LoSAI(rs=0.63-0.69,all P＜0.05),but there was no significant rank correlation with LoSDI(rs=0.27-0.33,P＞0.05).Conclusion Compared with the paired site,JLS active lesions had thinner subcutaneous tissue and stiffer skin.The higher the lesion skin stiffness,the more severe the JLS lesion activity.

Background: The widespread use of biological agents in recent years can improve the nutritional status of partial patients with Crohn's disease (CD) and change the course of the disease. Aims: To investigate the correlation between the nutritional risk and clinical and endoscopic features in CD patients treated with biological agents. Methods: Sixty⁃eight CD patients treated with biological agents from June 2021 to June 2022 at the First Affiliated Hospital of Anhui Medical University were enrolled. Nutritional risk screening was conducted by nutritional risk screening 2002 (NRS2002), and its correlation with clinical and endoscopic features was analyzed. Results: In 68 CD patients treated with biological agents, incidence of nutritional risk was 66.2%. There was statistically significant difference in incidence of nutritional risk in patients with different disease behaviors, SES⁃CD score and postoperative Rutgeerts score (all P<0.05). The incidence of nutritional risk was not correlated with age, lesion location, perianal lesion, CDAI score, and the interval between CD diagnosis and the use of biological agents (all P>0.05). Conclusions: The incidence of nutritional risk is high in CD patients treated with biological agents, and is correlated with CD disease behavior, SES ⁃ CD score, and postoperative Rutgeerts score.