Objective To investigate the bacterial community diversity in human Demodex mites, so as to provide insights into unraveling the role of human Demodex mites in them caused infectious diseases. Methods From June to July 2023, Demodex mites were collected from the faces of college students in a university in Wuhu City using the adhesive tape method, and the V4 region of 16S ribosomal RNA (16S rRNA) gene and the internal transcribed spacer (ITS) gene of nuclear ribosomal DNA were amplified on an Illumina PE250 high-throughput sequencing platform. Sequencing data were spliced according to the overlapping relations and filtered to yield effective sequences, and operational taxonomic units (OTUs) was clustered. The diversity index of obtained OUTs was analyzed, and the structure of the bacterial community was analyzed at various taxonomic levels. Results A total of 57 483 valid sequences were obtained using 16S rRNA gene sequencing, and 159 OUTs were classified according to similarity. Then, OUTs at a 97% similarity were included for taxonomic analyses, and the bacteria in Demodex mites belonged to 14 phyla, 20 classes, 51 orders, 72 families, and 94 genera. Proteobacteria was the dominant phylum, and Vibrio, Bradyrhizobium and Variovorax were dominant genera. A total of 56 362 valid sequences were obtained using ITS gene sequencing, and 147 OTUs were obtained, which belonged to 5 phyla, 17 classes, 34 orders, 68 families, and 93 genera and were annotated to Ascomycota, Basidiomycota and Chytridiomycota, with Ascomycota as the dominant phylum, and Alternaria alternata, Epicoccum, Penicillium, and Sarocladium as dominant genera. Conclusions There is a high diversity in the composition of bacterial communities in human Demodex mites, with multiple types of microorganisms and high species abundance.

Objective To investigate the potential active ingredients and the mechanism of Coreopsis tinctoria Nutt. in the prevention and treatment of hyperlipidemia. Methods Ultra-high performance liquid chromatography-Quadrupole-Exactive Orbitrap mass spectrometry (UPLC-Q-Exactive Orbitrap MS) was used to qualitatively analyze the fractions and blood components of flavones in Coreopsis tinctoria Nutt. The intersection targets of flavones in Coreopsis tinctoria Nutt. and hyperlipidemia were screened,and the protein-protein interaction network was constructed and analyzed by the STRING 12.0 database. Finally,the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used for enrichment analysis. Results A total of 25 compounds were detected from the flavones in Coreopsis tinctoria Nutt.,and their structures were identified,including ten chalcones,nine flavanones,four flavonols,one aurone,and one biflavone. The analysis of blood components showed that marein,flavanomarein,okanin,isookanin and 5,7,3',5'-tetrahydroxyflavanone-7-O-β-D-glucopyranoside were the main components of the flavones in Coreopsis tinctoria Nutt. in blood. Network pharmacological GO and KEGG enrichment analysis showed that the flavones in Coreopsis tinctoria Nutt. may regulate phosphatidylinositol 3-kinase/protein kinase B,tumor necrosis factor,hypoxia-inducible factor-1 signaling pathway and other signaling pathways in the regulation and prevention of hyperlipidemia. Conclusion Coreopsis tinctoria Nutt. can prevent and treat hyperlipidemia,and the mechanism may be related to the five blood components of the flavones in Coreopsis tinctoria Nutt.,including marein,flavanomarein,okanin,isookanin and 5,7,3',5'-tetrahydroxyflavanone-7-O-β-D-glucopyranoside.

Objective:To compare the detection method of hepatitis E virus (HEV) in simulated water samples, and to provide a reference for the detection of HEV in water.Methods:HEV fecal suspension was added to tap water or distilled water simulated water samples, and pretreatment was carried out by electropositive filter-organic eluent elution method (Method 1) to compare the extraction effect of the three nucleic acid extraction kits, A, B, and C. The simulated water samples were pre-treated by Method 1, 2 (electropositive filter-direct lysis method), 3 (tangential-flow ultrafiltration membrane-organic eluent elution method), and 4 (tangential-flow ultrafiltration membrane-direct lysis method) for pretreatment, A kit for nucleic acid extraction, Real time RT-PCR method for detection and comparison of the recovery rate; comparison of the recovery rate of different concentrations of HEV in simulated water samples; comparing the inhibitory effects of inhibitors in tap water samples on real time RT-PCR; and detection of HEV in different batches of tap water specimens.Results:Kit A nucleic acid extraction was better; the recoveries of method 1, 2, 3 and 4 were 7.31%, 39.88%, 6.85% and 64.88%, respectively, which showed a statistically significant difference in the recoveries ( F=114.069, P<0.001). The recoveries of method 4 with the addition of high, medium and low concentrations of HEV were 65.26%, 42.76% and 32.79%, respectively. The inhibition of all four pre-treatment method was less than 75%, which meets the requirements of ISO (15216-2∶2019). Twenty tap water specimens were tested for HEV and the result were negative. Conclusions:This study showed that the two membranes better recovered in combination with direct lysis, respectively; Methods 4 had a higher recovery in the detection of HEV in small volumes of distilled or tap water, but it was limited by the volume of water samples, turbidity, and so on. Suitable method can be selected for different water quality and laboratory conditions.

Objective:To optimize and compare method for hepatitis E virus (HEV) nucleic acid detection from pig liver, and provide technical references for HEV detection in animal viscera specimens.Methods:Three methods (PBS homogenization treatment, proteinase K treatment, chloroform extraction method) were used to pretreat and extract viral nucleic acid form pig liver, which was artificially contaminated with HEV fecal suspensions, and HEV RT-qPCR was used to compare the HEV recovery rate and inhibition rate. The optimized HEV method was applied to commercially available pig liver specimens, and HEV genotyping was performed on positive specimens.Results:The HEV recovery rate of PBS homogenization treatment, proteinase K treatment and chloroform extraction method was 9.88%, 0.19% and 17.28%, respectively. The recovery rate of proteinase K treatment was less than 1%, and it was discarded; t-test was performed to compare recovery rates of the other two methods, which showed statistically significant differences ( t=26.801, P<0.001), the chloroform extraction method had a higher recovery rate. The inhibition rates of the three methods were all less than 75%, within the range of the ISO/TS 15216-2∶2019 standard. Among 192 commercially available pig liver specimens, 17 specimens were detected positive for HEV RNA, with a nucleic acid positive rate of 8.85%; five specimens were successfully genotyped for HEV, all of which were genotype 4. Conclusions:The virus recovery effect was good when chloroform extraction method was used for pig liver pretreatment; moreover, this method could detect HEV RNA from commercially available pig livers, which indicate that it can be used for virus detection in food.

Objective To observe the value of prenatal ultrasound measuring fetal cauda equina nerve parameters for diagnosing tethered cord syndrome(TCS).Methods Forty six fetuses with TCS(TCS group)and 591 healthy fetuses(control group)were retrospectively enrolled.The length,area and angle of cauda equina nerve were measured with prenatal ultrasound and compared between groups,and the value for diagnosing TCS was analyzed.Results Significant differences of the length,area and angle of fetal cauda equina nerve were found between groups(all P＜0.05),with the area under the curve for diagnosing TCS of 0.924,0.809 and 0.972,respectively.Conclusion Prenatal ultrasound measuring fetal cauda equina nerve parameters had high value for diagnosing TCS.

OBJECTIVE To explore the clinical efficacy and safety of Ruyi jinhuang powder for external application combined with immune checkpoint inhibitors （ICIs） in the treatment of patients with advanced liver cancer complicated with dampness and heat syndrome of liver and gallbladder. METHODS All patients with advanced liver cancer complicated with dampness and heat syndrome of liver and gallbladder were admitted to our hospital from January 2018 to June 2022 and assigned into observation group and control group according to random number table method. Patients in the control group （n＝56） were treated with ICIs （Navulizumab injection/Sintilimab injection/Camrelizumab for injection） 200 mg， ivgtt， 21 days as a treatment cycle. Patients in the observation group （n＝56） were additionally treated with Ruyi jinhuang powder for external application， once a day， on the basis of control group. The therapeutic effects of 2 groups were compared after a treatment cycle. The levels of interleukin-6 （IL- 6）， matrix metalloproteinase-9 （MMP-9）， cyclooxygenase-2 （COX-2）， prostaglandin E2 （PGE2）， carbohydrate antigen 199 （CA199）， alpha-fetoprotein （AFP） and vascular endothelial growth factor （VEGF） in serum were compared between 2 groups before and after treatment. Karnofsky functional status （KPS） score， digital rating scale （NRS） score， total symptom score of traditional Chinese medicine， and the occurrence of adverse reactions were recorded for both groups of patients. RESULTS After treatment， the levels of IL-6， MMP-9， COX-2， PGE2， CA199， AFP， VEGF， NRS score and total symptom score of traditional Chinese medicine in observation group were significantly lower than control group （P＜0.05）， KPS score was significantly higher than the control group （P＜0.05）. The zypp-04） total effective rate and remission rate of the observation group were 64.29% and 80.36%， those of control group were 60.71% and 73.21%. There was no statistical significance between two groups （P＞0.05）. The adverse drug reactions of both groups were mainly nausea and vomiting， liver function injury， fever hlshli@yeah.net and so on； the incidence of adverse reaction in observation group was significantly lower than that of control group （P＜0.05）. CONCLUSIONS In patients with advanced liver cancer complicated with dampness and heat syndrome of liver and gallbladder， the combination of Ruyi jinhuang powder for external application and ICIs can help inhibit the secretion of pain mediators， regulate vascular endothelial function， reduce the inflammatory response， promote the recovery of cardiopulmonary function， improve clinical efficacy and has good safety.

Objective: The study aimed to investigate the relationship between academic stress and extracurricular physical exercise among college students, with the goal of providing evidence and reference for effectively alleviating academic stress. Methods: A stratified cluster random sampling method was employed, along with the Chinese College Student Psychological Health Screening Scale and a self designed questionnaire. In January 2023, an online survey was conducted among 1 108 college students from eight colleges and universities in Jiangsu Province. Statistical analyses included independent samples t test, one way analysis of variance (ANOVA) and the optimal scale regression model. Results: It revealed that 50.8% of students worried about falling behind their peers in their studies, 51.7% expressed concerns about exams, 21.7% exerted significant effort but still struggled to keep up with the pace of learning, and 24.9% felt difficulties in their studies. The overall index of academic stress was higher among undergraduate institutions, first year students, female students, and urban residents ( t/F =3.66, 10.96, -5.46, 3.52, P <0.01). The average time of weekly extracurricular physical exercise for students was (2.35±2.18)h, and there were significant differences by gender, only child status, and grade groups( t/F =2.35, 4.66, 7.09, P <0.05). The optimal scale regression model analysis ( R 2=0.17, F=32.03, P <0.01) revealed that the regression coefficient and partial correlation coefficient between academic stress and extracurricular physical exercise time were both -0.33( P <0.01). The importance of extracurricular physical exercise in the model was 0.66, with a tolerance value of 0.97. The regression coefficients and partial correlation coefficients for academic stress and extracurricular physical exercise time were -0.35, -0.35 for undergraduate students, and -0.31, -0.32 for junior college students ( P < 0.01 ). Conclusion A significant number of college students experience varying degrees of academic stress, and their engagement in extracurricular physical exercise is insufficient. There is a negative association between academic stress and extracurricular physical activity time among college students. Universities should encourage students to increase extracurricular physical activity time to reduce academic stress and promote college students mental health.

Objective:To investigate the diagnosis, treatment, clinical characteristics and potential high-risk factors of cerebral venous sinus thrombosis (CVST) during the treatment of acute lymphoblastic leukemia (ALL) with pegaspargase.Methods:The medical history, diagnosis and treatment process, laboratory examination and imaging examination results of 3 ALL patients with CVST during pegaspargase treatment in the First Affiliated Hospital of Suzhou University in March and November 2021 and September 2022 were retrospectively analyzed, and the relevant literature was reviewed.Results:Three patients were all female, with the aged between 15 and 35 years old, including 2 cases of B-ALL and 1 case of T-ALL. All patients developed nervous system symptoms after pegaspargase chemotherapy, and were diagnosed as CVST by imaging examination. During the pegaspargase treatment, 2 patients took norethisterone, and 1 patient underwent induced labor and curettage. The levels of sexual hormones in the 3 patients had non-physiological changes. The main CVST lesions were located in the superior sagittal sinus, transverse sinus and sigmoid sinus. One patient had cerebral hemorrhage at the same time. When thrombus occurred, the fibrinogen (Fib), antithrombin Ⅲ (AT Ⅲ) activity, protein C activity and protein S activity of the patients were significantly lower than those before, D-dimer was significantly higher, and lupus anticoagulant and anticardiolipin antibody were negative. The thrombosis treatment was mainly anticoagulation, and 1 patient underwent thrombolysis. Two patients had no sequelae of nervous system, and 1 patient had the sequelae of muscle weakness.Conclusions:Patients with ALL should be alert to the occurrence of CVST when they have nervous system symptoms during pegaspargase chemotherapy. The diagnosis of CVST mainly depends on cranial imaging. Anticoagulation is the main thrombosis treatment, thrombolysis and interventional thrombectomy are feasible for some patients, with few neurological sequelae. The use of second-generation progesterone drugs and the non-physiological fluctuation of sex hormones may be the potential risk factors of CVST.

Objective:The two detection method for the detection of hepatitis A virus (HAV) in strawberries were optimized and compared to select the best detection method for the detection of hepatitis A virus in strawberries.Methods:Different concentrations of HAV were inoculated on the surface of known negative frozen strawberry specimens, the concentration of beef extract powder in alkaline elution-PEG concentration method was optimized, the optimal nucleic acid extraction kit was selected, the optimal lysis buffer volume in direct lysis method was optimized, and the recovered viral load was detected by Real-time fluorescence quantitative RT-PCR. SPSS26.0 was used to statistically analyze the data, and the optimized two method were used for the detection of actual specimens.Results:The concentration of beef extract powder by the optimized alkaline elution-PEG concentration method was selected at 3%, and the viral nucleic acid extraction kit was selected as kit B. Six ml of lysis buffer was selected for the optimized direct lysis method. The recovery rates of HAV virus by alkaline elution-PEG concentration and direct lysis were compared with the HAV addition levels, and the HAV virus recovery rates of the two method were 21.50±1.06% and 5.82±0.01%, respectively, and the result showed that the differences were statistically significant. A total of 60 strawberry specimens from four regions were tested at the same time, and the result were all negative.Conclusions:The optimized alkaline elution-PEG concentration method has higher sensitivity and is more suitable for the detection of HAV in strawberry specimens.

Objective:To establish a digital droplet RT-PCR(dRT-PCR) method for Hepatitis A virus (HAV), and compare it with Real time RT-PCR(RT-qPCR) method, and select the best method for detecting hepatitis A virus in strawberry.Methods:Extract HAV vaccine RNA, optimize the reaction conditions of dRT-PCR and evaluate its specificity; Alkaline elution -PEG concentration method was used to extract nucleic acid from strawberry samples. At the same time, dRT-PCR and RT-qPCR method were used to detect the sensitivity and inhibition rate of HAV vaccine RNA in pure water and strawberry matrix, and the recovery rate of HAV in artificially contaminated strawberry was compared, which was applied to the detection of commercially available samples.Results:The optimal annealing temperature for dRT-PCR reaction was 60 ℃, and the optimal concentrations of primers and probes were 0.4 μmol/L、0.4 μmol/L and 0.2 μmol/L, with good specificity. There is no significant difference in sensitivity between the two method in detecting HAV vaccine RNA in pure water and strawberry matrix. The inhibition rate of dRT-PCR is low. The recovery rates of dRT-PCR and RT-qRCR in the detection of strawberry samples contaminated with HAV at higher concentrations were 12.90±0.006% and 30.12±0.02%, respectively. The recovery rates of lower concentrations of HAV contaminated strawberry samples were 18.27±0.07% and 10.85±0.03%, respectively, and the difference was statistically significant ( P<0.05). When strawberry samples on the market were tested, the result of both method were negative. Conclusions:The sensitivity of dRT-PCR method established in this study is not significantly different from that of RT-qPCR in detecting HAV RNA in different substrates, but dRT-PCR has good tolerance to PCR reaction inhibitors and high recovery rate when detecting low concentration HAV. Both detection method can be used for quantitative detection of hepatitis A virus in strawberry, and can be selected according to the actual situation.