ObjectiveTo investigate the antidepressant effects and mechanisms of total flavonoids from Cuscutae Semen （TFCC） in the mouse model of chronic unpredictable mild stress （CUMS）. MethodsFifty male 4-week-old ICR mice were randomized into five groups （n=10 per group）： blank control， model， Cuscutae Semen decoction （10.2 g·kg^-1·d^-1）， paroxetine （2.6 mg·kg^-1·d^-1）， and TFCC （173.2 mg·kg^-1·d^-1）. The other groups except the blank control group underwent chronic unpredictable mild stress （CUMS） for 4 weeks. Behavioral assessments were conducted post-modeling. Then， the model group received distilled water （10 mL·kg^-1·d^-1）， while treatment groups were administrated with respective agents via oral gavage （10 mL·kg^-1） for 4 weeks. Depression-like behaviors were evaluated by the sucrose preference test （SPT）， forced swimming test （FST）， and tail suspension test （TST）. Hippocampal neuronal morphology was observed via hematoxylin-eosin staining， and apoptosis in the brain tissue was assessed via terminal- deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling （TUNEL）. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the hippocampal levels of inflammatory cytokines ［interleukin （IL）-1β， IL-6， and TNF-α）］ and neurotransmitters ［5-hydroxytryptamine （5-HT）， dopamine （DA）， and brain-derived neurotrophic factor （BDNF）］， while the reactive oxygen species （ROS） levels were quantified via the DCFH-DA probe. Real-time PCR was performed to measure the mRNA levels of NOD-like receptor protein 3 （NLRP3）， apoptosis-associated Speck-like protein containing a CARD （ASC）， cysteinyl aspartate-specific proteinase-1 （Caspase-1）， IL-1β， and inducible nitric oxide synthase （iNOS）. Western blot was employed to evaluate the protein levels of NLRP3， ASC， Caspase-1， uncoupling protein 2 （UCP2）， and thioredoxin-interacting protein （TXNIP）. ResultsCompared with the blank control group， the model group exhibited weight loss （P<0.01）， reduced sucrose preference （P<0.01）， prolonged immobility time in FST and TST （P<0.01）， neuron disarrangement with nuclear pyknosis in hippocampal CA3 region， increased apoptosis in the brain tissue， elevated levels of IL-1β， IL-6， and TNF-α （P<0.01）， declined levels of 5-HT， DA， and BDNF （P<0.01）， increased ROS accumulation （P<0.01）， upregulated mRNA levels of NLRP3， ASC， Caspase-1， IL-1β， and iNOS （P<0.01）， down-regulated protein level of UCP2 （P<0.01）， and up-regulated protein levels of NLRP3， ASC， Caspase-1， and TXNIP （P<0.01）. Compared with the model group， the interventions restored sucrose preference （P<0.01）， shortened immobility time （P<0.01）， repaired hippocampal neuronal structure， reduced apoptosis， lowered the levels of inflammatory cytokines （P<0.01）， restored the levels of neurotransmitters （P<0.01）， alleviated ROS accumulation （P<0.01）， downregulated the mRNA levels of NLRP3， ASC， Caspase-1， IL-1β， and iNOS （P<0.01）， upregulated the protein level of UCP2 （P<0.01）， and reduced the protein levels of NLRP3， ASC， Caspase-1， and TXNIP （P<0.01）. Moreover， TFCC outperformed Cuscutae Semen decoction in ameliorating depressive behaviors. TFCC excelled in neuronal repair， neurotransmitter regulation， anti-inflammatory effects， and modulation of the UCP2/TXNIP/NLRP3 pathway （P<0.05）. ConclusionTFCC modulates the hippocampal UCP2/TXNIP/NLRP3 pathway to inhibit inflammasome activation， reduce oxidative stress， restore neurotransmitters， thus suppressing neuronal apoptosis and promoting the rearrangement and morphology recovery of hippocampal cells. It outperforms Cuscutae Semen decoction in the antidepressant efficacy.

ObjectiveTo investigate the antidepressant effects and mechanisms of total flavonoids from Cuscutae Semen （TFCC） in the mouse model of chronic unpredictable mild stress （CUMS）. MethodsFifty male 4-week-old ICR mice were randomized into five groups （n=10 per group）： blank control， model， Cuscutae Semen decoction （10.2 g·kg^-1·d^-1）， paroxetine （2.6 mg·kg^-1·d^-1）， and TFCC （173.2 mg·kg^-1·d^-1）. The other groups except the blank control group underwent chronic unpredictable mild stress （CUMS） for 4 weeks. Behavioral assessments were conducted post-modeling. Then， the model group received distilled water （10 mL·kg^-1·d^-1）， while treatment groups were administrated with respective agents via oral gavage （10 mL·kg^-1） for 4 weeks. Depression-like behaviors were evaluated by the sucrose preference test （SPT）， forced swimming test （FST）， and tail suspension test （TST）. Hippocampal neuronal morphology was observed via hematoxylin-eosin staining， and apoptosis in the brain tissue was assessed via terminal- deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling （TUNEL）. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the hippocampal levels of inflammatory cytokines ［interleukin （IL）-1β， IL-6， and TNF-α）］ and neurotransmitters ［5-hydroxytryptamine （5-HT）， dopamine （DA）， and brain-derived neurotrophic factor （BDNF）］， while the reactive oxygen species （ROS） levels were quantified via the DCFH-DA probe. Real-time PCR was performed to measure the mRNA levels of NOD-like receptor protein 3 （NLRP3）， apoptosis-associated Speck-like protein containing a CARD （ASC）， cysteinyl aspartate-specific proteinase-1 （Caspase-1）， IL-1β， and inducible nitric oxide synthase （iNOS）. Western blot was employed to evaluate the protein levels of NLRP3， ASC， Caspase-1， uncoupling protein 2 （UCP2）， and thioredoxin-interacting protein （TXNIP）. ResultsCompared with the blank control group， the model group exhibited weight loss （P<0.01）， reduced sucrose preference （P<0.01）， prolonged immobility time in FST and TST （P<0.01）， neuron disarrangement with nuclear pyknosis in hippocampal CA3 region， increased apoptosis in the brain tissue， elevated levels of IL-1β， IL-6， and TNF-α （P<0.01）， declined levels of 5-HT， DA， and BDNF （P<0.01）， increased ROS accumulation （P<0.01）， upregulated mRNA levels of NLRP3， ASC， Caspase-1， IL-1β， and iNOS （P<0.01）， down-regulated protein level of UCP2 （P<0.01）， and up-regulated protein levels of NLRP3， ASC， Caspase-1， and TXNIP （P<0.01）. Compared with the model group， the interventions restored sucrose preference （P<0.01）， shortened immobility time （P<0.01）， repaired hippocampal neuronal structure， reduced apoptosis， lowered the levels of inflammatory cytokines （P<0.01）， restored the levels of neurotransmitters （P<0.01）， alleviated ROS accumulation （P<0.01）， downregulated the mRNA levels of NLRP3， ASC， Caspase-1， IL-1β， and iNOS （P<0.01）， upregulated the protein level of UCP2 （P<0.01）， and reduced the protein levels of NLRP3， ASC， Caspase-1， and TXNIP （P<0.01）. Moreover， TFCC outperformed Cuscutae Semen decoction in ameliorating depressive behaviors. TFCC excelled in neuronal repair， neurotransmitter regulation， anti-inflammatory effects， and modulation of the UCP2/TXNIP/NLRP3 pathway （P<0.05）. ConclusionTFCC modulates the hippocampal UCP2/TXNIP/NLRP3 pathway to inhibit inflammasome activation， reduce oxidative stress， restore neurotransmitters， thus suppressing neuronal apoptosis and promoting the rearrangement and morphology recovery of hippocampal cells. It outperforms Cuscutae Semen decoction in the antidepressant efficacy.

Objective To explore the efficacy of blunt separation method in midline catheter intubation in elderly patients with coagulation dysfunction.Methods A total of 80 elderly patients with coagulation dysfunction were selected in the convenience sampling method from October 2022 to April 2023 in our hospital,and they were randomly divided into an experimental(blunt)group and a control(routine)group,with 40 patients in each group.The differences in the degree of bleeding and exudation at the puncture site,the pain score and the incidence of complications(including bleeding and exudation,phlebitis,symptomatic catheter-related thrombus,catheter blockage,catheter pulling-off)were compared between 2 groups.Results In the experimental group,the degree of bleeding and exudation at the puncture point immediately after the operation,degree of bleeding and exudation at the puncture point 24 hours after the operation,pain score 1 day after the catheterization,pain score 3 days after the catheterization,incidence of bleeding and exudation,total incidence of complications and maintenance times were significantly lower than these in the control group(P＜0.05).In terms of the pain score immediately after the operation,pain score 5 days after the operation,incidence of phlebitis,incidence of symptomatic catheter-related thrombosis,incidence of catheter blockage,incidence of catheter pulling-off,incidence of catheter related skin injury,incidence of unplanned extubation,success rate of one-time sheath delivery and the indwelling time,the differences between the experimental group and control group were not significant(P＜0.05).Conclusion The application of blunt separation method in midline catheter indwelling can significantly reduce the incidence and degree of bleeding at the puncture point,decrease the maintenance times and relieve the pain in elderly patients with coagulation dysfunction.

Objective:To study the epidemiological and genetic characteristics of coxsackievirus B3 (CVB3) circulating in Guangdong Province from 2008 to 2021.Methods:This study collected the specimens of hand, foot, and mouth disease (HFMD) cases from 2008 to 2021 that were positive for other enteroviruses except for enterovirus 71 (EV71), coxsackievirus A16 (CVA16), and CVA6, as well as the specimens of herpangina and neonatal infection cases from 2020 to 2021. Enteroviruses in these specimens were detected and their types were identified. CVB3 strains were isolated and the entire VP1 sequences of CVB3 strains were amplified and sequenced. The genetic features of CVB3 strains were analyzed using DNAStar 7.1 and MEGA 6.06 software packages.Results:Among 3 484 HFMD cases positive for other enteroviruses from 2008 to 2021, CVB3-positive cases accounted for 1.6% (57/3 484); among 560 cases of herpangina from 2020 to 2021, CVB3-positive cases accounted for 2.1% (12/560); one neonatal infection case in 2021 was positive for CVB3. CVB3-positive cases accounted for 67.1% (47/70) in 2021 and 18.6% (13/70) in 2020, while there were less than five cases in other years. Forty-eight CVB3 strains were isolated and the entire VP1 sequences of 26 CVB3 strains were obtained. Phylogenetic analysis indicated that the CVB3 strains could be divided into eight genotypes (A-H) and the strains of genotypes A, D and E were prevalent in the Chinese mainland. The 26 CVB3 strains isolated in Guangdong Province shared 80.2%-100.0% nucleotide homology, and belonged to two genotypes of D and E, with genotype D prevalent from 2008 to 2017 and genotype E prevalent from 2020 to 2021.Conclusions:CVB3 is prevalent sporadically in Guangdong Province from 2008 to 2017, but the epidemic intensity increased during 2020 and 2021. CVB3 strains of genotypes D and E are prevalent in Guangdong Province during 2008 to 2021, with genotype E being the prevalent genotype during 2020 and 2021.

Objective To investigate changes and clinical significance of serum microRNA-155(miR-155)and microRNA-205-5p(miR-205-5p)levels in patients with hepatitis C virus(HCV)infection.Methods A total of 141 patients with HCV infection were collected and divided into the HCV-1b group(92 cases)and the non HCV-1b group(49 cases)according to their genotype,and another 141 healthy volunteers who underwent physical examinations in our hospital during the same period were collected as the control group.The general information,biochemical indicators and serum levels of miR-155 and miR-205-5p were compared between the three groups.According to the grading of hepatitis activity,HCV infected patients were classified into the G0,G1,G2,G3 and G4 groups,and serum levels of miR-155 and miR-205-5p were compared between the five groups.Receiver operating characteristics(ROC)curve was used to analyze clinical values of serum miR-155 and miR-205-5p in diagnosing hepatitis activity of G1-G4 in patients with HCV infection.Results The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and total bilirubin(TBIL)were obviously higher in the HCV-1b group and the non HCV-1b group than those in the control group,while the level of ALB was obviously lower than that in the control group(P＜0.05).The serum level of miR-155 was increased in turn in the control group,the non-HCV-1b group and the HCV-1b group,and the serum level of miR-205-5p was decreased in turn(P＜0.05).The serum level of miR-155 increased in turn with the increased grading of hepatitis activity,while the serum miR-205-5p level decreased in turn(P＜0.05).The area under curve(AUC)values of serum miR-155,miR-205-5p and their combination in diagnosing hepatitis activity of G1-G4 in HCV infected patients were 0.855,0.793 and 0.913,respectively,and the combination of the two is superior to each individual diagnosis(P＜0.05).Conclusion The serum miR-155 level increases and miR-205-5p level decreases in HCV infected patients.The combination of the two has high efficacy in diagnosing hepatitis activity of G1-G4 in HCV infected patients.

Objective:To investigate the association between intestinal colonization of segmented filamentous bacteria (SFB) and the risk of rotavirus infection, and the possible mechanisms by which SFB resist rotavirus infection.Methods:This case-control study enrolled 50 children aged 0 to 5 years who present to the outpatient Department of Children′s Hospital, Zhejiang University School of Medicine with diarrhea and positive stool tests for rotavirus. The children were divided into rotavirus enteritis group and control group consisting of 55 children with non-gastrointestinal and non-infectious surgical diseases.The age and sex composition of the two groups was matched. The DNA of the fecal flora was extracted and SFB was detected by real-time fluorescence quantitative PCR analysis. The children in the rotavirus enteritis group and the control group were subgrouped by age and sex to analyze the differences in SFB positivity rates between different groups, and further compare and analyze the differences in SFB positivity rates between these two groups of children in the ≤2 years old subgroup and the >2-5 years old subgroup. Neutralization test was performed with p3340 protein and rotavirus to determine the relationship between rotavirus infection rate and p3340 concentration in Vero cells. χ2 test or Fisher′s exact probability method was used for comparison between the two groups. Results:There were 50 children in the rotavirus enteritis group with an age of (1.7±0.9) years, and 55 children in the control group with an age of (1.8±1.1) years. The positive rate of SFB in children with rotavirus enteritis showed a declining trend across ages groups, with the highest rate of 10/14 in the ≤1 year old group, followed by 67% (14/21) in the >1-2 years old group, 9/15 in the >2-5 years old group, and there was no statistically significant difference ( P=0.867). The positive rate of SFB in the control group was 12/15 in the ≤1 year old group, 95% (19/20) in the >1-2 years old group, 50% (10/20) in the >2-5 years old group, with statistical significance ( P=0.004). The positive rate of SFB in children with rotavirus enteritis was 74% (20/27) in males and 56% (13/23) in females ( χ2=1.71, P=0.192). In the control group, it was 79% (22/28) in males and 70% (19/27) in females ( χ2=0.49, P=0.485). The positive rate of SFB was 66% (33/50) in the rotavirus enteritis group and 75% (41/55) in the control group, with no statistically significant ( χ2=0.56, P=0.454). In the children ≤2 years old, the SFB positivity rate was 69% (24/35) in the rotavirus enteritis group and 89% (31/35) in the control group, with a statistically significant difference ( χ2=4.16, P=0.041). However, in the children >2-5 years old, no statistically significant difference was observed, with the positive rate of SFB being 9/15 in the rotavirus enteritis group and 50% (10/20) in the control group ( P=0.734). Pearson correlation analysis revealed a negative correlation between rotavirus infection and SFB positivity ( r=-0.87, P<0.001). As the concentration of the p3340 specific protein increased, the luminescence intensity of the luciferase in the Vero cells, which were suitable for cultivating rotavirus, exhibited a decreasing trend ( F=4.17, P=0.001). Conclusions:SFB colonization in infants less than 2 years old is associated with a reduced risk of rotavirus infection. Cloning of specific SFB functional protein p3340 neutralizes rotavirus infection of Vero cells, and this mechanism of targeting rotavirus infection differs from the common antiviral mechanism.

Methamphetamine (METH) abuse is associated with significant neurotoxicity, high addiction potential, and behavioral abnormalities. Recent studies have identified a connection between the gut microbiota and METH-induced neurotoxicity and behavioral disorders. However, the underlying causal mechanisms linking the gut microbiota to METH pathophysiology remain largely unexplored. In this study, we employed fecal microbiota transplantation (FMT) and antibiotic (Abx) intervention to manipulate the gut microbiota in mice administered METH. Furthermore, we supplemented METH-treated mice with short-chain fatty acids (SCFAs) and pioglitazone (Pio) to determine the protective effects on gut microbiota metabolism. Finally, we assessed the underlying mechanisms of the gut-brain neural circuit in vagotomized mice. Our data provide compelling evidence that modulation of the gut microbiome through FMT or microbiome knockdown by Abx plays a crucial role in METH-induced neurotoxicity, behavioral disorders, gut microbiota disturbances, and intestinal barrier impairment. Furthermore, our findings highlight a novel prevention strategy for mitigating the risks to both the nervous and intestinal systems caused by METH, which involves supplementation with SCFAs or Pio.

Methamphetamine (Meth) abuse can cause serious mental disorders, including anxiety and depression. The gut microbiota is a crucial contributor to maintaining host mental health. Here, we aim to investigate if microbiota participate in Meth-induced mental disorders, and the potential mechanisms involved. Here, 15 mg/kg Meth resulted in anxiety- and depression-like behaviors of mice successfully and suppressed the Sigma-1 receptor (SIGMAR1)/BDNF/TRKB pathway in the hippocampus. Meanwhile, Meth impaired gut homeostasis by arousing the Toll-like receptor 4 (TLR4)-related colonic inflammation, disturbing the gut microbiome and reducing the microbiota-derived short-chain fatty acids (SCFAs). Moreover, fecal microbiota from Meth-administrated mice mediated the colonic inflammation and reproduced anxiety- and depression-like behaviors in recipients. Further, SCFAs supplementation optimized Meth-induced microbial dysbiosis, ameliorated colonic inflammation, and repressed anxiety- and depression-like behaviors. Finally, Sigmar1 knockout (Sigmar1^-/-) repressed the BDNF/TRKB pathway and produced similar behavioral phenotypes with Meth exposure, and eliminated the anti-anxiety and -depression effects of SCFAs. The activation of SIGMAR1 with fluvoxamine attenuated Meth-induced anxiety- and depression-like behaviors. Our findings indicated that gut microbiota-derived SCFAs could optimize gut homeostasis, and ameliorate Meth-induced mental disorders in a SIGMAR1-dependent manner. This study confirms the crucial role of microbiota in Meth-related mental disorders and provides a potential preemptive therapy.

Objective:The investigation and research on the application status of Hepatic Venous Pressure Gradient (HVPG) is very important to understand the real situation and future development of this technology in China.Methods:This study comprehensively investigated the basic situation of HVPG technology in China, including hospital distribution, hospital level, annual number of cases, catheters used, average cost, indications and existing problems.Results:According to the survey, there were 70 hospitals in China carrying out HVPG technology in 2021, distributed in 28 provinces (autonomous regions and municipalities directly under the central Government). A total of 4 398 cases of HVPG were performed in all the surveyed hospitals in 2021, of which 2 291 cases (52.1%) were tested by HVPG alone. The average cost of HVPG detection was (5 617.2±2 079.4) yuan. 96.3% of the teams completed HVPG detection with balloon method, and most of the teams used thrombectomy balloon catheter (80.3%).Conclusion:Through this investigation, the status of domestic clinical application of HVPG has been clarified, and it has been confirmed that many domestic medical institutions have mastered this technology, but it still needs to continue to promote and popularize HVPG technology in the future.

Objective To evaluate the efficacy of transcatheter arterial embolization (TAE) in treating oronasal cavity hemorrhage,and to discuss the the occurrence and prevention of complications.Methods The clinical data of 121 patients with refractory and fatal oronasal cavity hemorrhage,who were admitted to authors' hospital during the period from December 2005 to October 2013 to receive treatment,were retrospectively analyzed.A total of 116 patients were treated with TAE,and these patients were followed up for 1-3 months to evaluate the embolization effect and the occurrence of procedure-related complications was analyzed.Results Of the 116 patients,complete control of bleeding after TAE was achieved in 96 (82.7％),rebleeding within one week after TAE was seen in 19 (16.4％) and the bleeding was controlled by medication,and in the remaining one (0.9％) re-bleeding occurred within one week after TAE and embolization therapy had to be carried out again.No obvious complications occurred in 77 patients (66.4％);maxillofacial pain and numbness,low fever,limitation of mouth opening and other mild complications were observed in 35 patients (30.1％);one patient (0.9％) developed facial skin necrosis and severe headache;and 3 patients (2.6％) showed stroke symptoms due to cerebral embolism.Conclusion For the treatment of refractory and fatal oronasal cavity hemorrhage,TAE can quickly and effectively achieve the purpose of hemostasis;careful selection of proper embolization material based on the the different causes of bleeding and the responsible blood vessels is the key to ensure a successful TAE.The common postoperative complications include postembolization syndrome,local ischemia,local necrosis caused by peripheral ischemia;the main serious complications are skin necrosis of maxillofacial region and cerebral infarction caused by ectopic embolization.