Objective To explore the anti-depression mechanism of Kai-Xin-San(KXS)via regulation of neurogenesis in hippocampus of depression-like mice.Methods The extracts of KXS were prepared and the anti-depression effects of KXS were evaluated by behavioral tests on chronic unpredictable mild stress(CUMS)induced depression-like mice.Evaluating depression-like behavior in CUMS mice through sucrose preference test,forced swimming test,tail suspension test,and other methods.Neurogenesis in hippocampus were determined by immunofluorescence assay.In addition,effects of KXS on regulating nestin expression and Wnt/b-catenin signaling pathway were explored by western blotting analysis.Amounts of cortisol,corticotropin-releasing factor(CRF),adrenocorticotropic hormone(ACTH),brain-derived neurotrophic factor(BDNF)and nerve growth factor(NGF)were determined by ELISA tests.Mouse primary neural stem cells(NSC)was used to evaluate the effect of KXS on promoting its proliferation by immunofluorescence assay.In addition,effects of KXS on regulating nestin and Wnt/β-catenin signaling pathway were also explored by Western blotting analysis.Results KXS significantly ameliorated the depression-like behaviors in presence of increased sucrose preference rate and decreased immobile time of tail suspension and forced swimming.KXS significantly promoted the neurogenesis in the hippocampus and expressions of nestin,reduced the expressions of cortisol,CRF,ACTH,increased the expressions of BDNF,NGF,and regulated Wnt/β-catenin signaling pathway.KXS also promoted the proliferation of NSCs and expressions of nestin,enhanced the translocation of b-catenin into nucleus,and regulated the expressions of proteins of Wnt/β-catenin signaling pathway.Conclusion KXS promoted neurogenesis in hippocampus and regulated Wnt/β-catenin pathway,which might contribute to its antidepressant effect.

Objective To investigate the beneficial effect of Kai-Xin-San combined with fluoxetine in improving depression-like behaviors on chronic unpredictable mild stress(CUMS)induced depression model mice.Methods The present study aimed to assess the potential of Kai-Xin-San in combination with fluoxetine to ameliorate depression-like behaviors in a CUMS induced mouse depression model.Behavioral tests,such as the sucrose preference test were employed to evaluate the efficacy of the treatment.Additionally,the levels of suppressed stress factors were measured using the ELISA method.The morphology of hippocampal tissue was evaluated using the HE staining method,Nissl Staining and TUNEL staining methods.Furthermore,western blotting analysis was utilized to determine the expression levels of proteins such as Caspase-3,and Caspase-9.Results The co-administration of Kai-Xin-San and fluoxetine resulted in a significant increase in sucrose preference rate in model mice.This effect was comparable to that of fluoxetine alone at the standard clinical dose.Furthermore,the combination treatment up-regulated the levels of suppressed stress factors,reduced the apoptosis of hippocampus induced by depression and regulated the apoptosis signaling pathway in hippocampus.Conclusion The combination of Kai-Xin-San and fluoxetine has been shown to be an effective treatment for depression-like behavior in animal models,resulting in a reduction in the required clinical dosage of fluoxetine.This effect may be attributed to the up-regulation of neurotransmitter expression,inhibition of stress axis activation,and central nervous inflammation.

OBJECTIVE To evaluate the ameliorative effect of Juanbi Tongluo Oral Liquid on sciatic neuronal apoptosis in Type 2 diabetic model mice.METHODS The Type 2 diabetes mouse model was established by feeding with high-fat and high-sugar diet combined with intraperitoneal injection of streptozotocin(STZ).The mice were treated with metformin(200 mg·kg-1·d-1),low dose(3.9 g·kg-1·d-1)and high dose(7.8 g·kg-1·d-1)Juanbi Tongluo Oral Liquid for 35 days.The latency of response to thermal stimu-lation was detected by hot plate,and the values of blood glucose insulin and glycosylated hemoglobin were determined.Biochemical kits were used to detect the expression of serum total cholesterol(T-CHO),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px)and oxidation product malondialdehyde(MDA).The expression of tumor cytokine α(TNF-α),interleukin(IL)-1β,IL-6 and IL-10 in serum of mice were detected by ELISA method;the injury of sciatic nerve of model mice was detected by HE staining;the apoptosis of sciatic nerve was detected by TUNEL method;and the expression of neurofilament protein NF-L,apoptosis(cleaved Caspase-3,Caspase-3)and oxidative stress(Nrf2,HO-1)signal pathway proteins in sciatic nerve of model mice were detected by Western blot method.RESULTS High-dose Juanbi Tongluo Oral Liquid shortened the latent period of heat pain response in model mice(P<0.01);downregulated fasting blood glucose and glycated hemoglobin(P<0.01),and upregulated fasting plasma insulin in model mice(P<0.01);downregulated serum T-CHO,TG,and LDL-C levels(P<0.01),upregulated HDL-C levels(P<0.01);downregulated serum pro-inflammatory factors TNF-α,IL-1β and IL-6 levels(P<0.05),upregulated the anti-inflammatory cyto-kine IL-10 level(P<0.01);inhibited sciatic nerve structural damage and apoptosis(P<0.05);downregulated the ratio of cleaved Caspase-3 to Caspase-3 in the apoptosis pathway(P<0.01);upregulated the expression of neurofilament proteins NF-L and NF-H in sciatic nerve tissue(P<0.05,P<0.01);and upregulated the expression of antioxidant stress proteins Nrf2 and HO-1(P<0.01).CONCLUSION Juanbi Tongluo Oral Liquid can improve sciatic neuronal apoptosis of Type 2 diabetic mice,which may be related to its effect on improving oxidative stress and inflammatory stress.

Objective To prepare a dissolvable microneedle(DMN)with a tip-layer loaded with hyaluronic acid(HA)modified sinomenine hydrochloride liposomes(HA-SMH-Lip),as well as characterize,evaluate its in vitro transdermal permeability,cellular uptake ability,and anti-inflammatory ability.Methods HA-SMH-Lip-DMNs were prepared by a two-step casting method,and the drug loading capacity was determined using HPLC.The morphology,skin permeation properties and in vitro transdermal ability were investigated by scanning electron microscopy,puncture assay and Franz diffusion cell method.Fluorescent microneedles were prepared by replacing HA-SMH-Lip with fluorescein isothiocyanate liposomes(HA-FITC-Lip/FITC-Lip).The uptake behavior of inflammation cells on HA-FITC-Lip-DMNs/FITC-Lip-DMNs was investigated using a flow cytometer and a fluorescence microscope.To evaluate the anti-inflammatory activity of HA-SMH-Lip-DMNs,the levels of inflammatory factors including nitric oxide(NO),tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β),and IL-10 in cell supernatants were measured using an ELISA kit.Results The prepared HA-SMH-Lip-DMNs have uniform shape and size,integral and visually pleasing array,and an average drug loading of(114.01±1.04)μg.Additionally,they have good puncture ability.The results of in vitro transdermal experiments showed that the accumulated amounts of HA-SMH-Lip-DMNs were(101.47±2.91)μg·cm-2 at 36 hours.Its transdermal ability was better than that of the SMH solution group and SMH liposome group.In vitro cellular uptake results indicated that HA-FITC-Lip-DMNs were more effectively taken up by RAW 264.7 cells(P＜0.01).Compared to the model group,HA-SMH-Lip-DMNs group significantly reduced TNF-α,IL-1β,and NO levels while increase IL-10 levels(P＜0.01).Conclusion The prepared HA-SMH-Lip-DMNs have a complete and beautiful morphology with excellent cellular uptake capability,remarkable in vitro transdermal performance,and potent anti-inflammatory properties.HA-SMH-Lip-DMNs are expected to become a new type of transdermal drug delivery system.

Ameloblastoma is a benign tumor characterized by locally invasive phenotypes,leading to facial bone destruction and a high recurrence rate.However,the mechanisms governing tumor initiation and recurrence are poorly understood.Here,we uncovered cellular landscapes and mechanisms that underlie tumor recurrence in ameloblastoma at single-cell resolution.Our results revealed that ameloblastoma exhibits five tumor subpopulations varying with respect to immune response(IR),bone remodeling(BR),tooth development(TD),epithelial development(ED),and cell cycle(CC)signatures.Of note,we found that CC ameloblastoma cells were endowed with stemness and contributed to tumor recurrence,which was dominated by the EZH2-mediated program.Targeting EZH2 effectively eliminated CC ameloblastoma cells and inhibited tumor growth in ameloblastoma patient-derived organoids.These data described the tumor subpopulation and clarified the identity,function,and regulatory mechanism of CC ameloblastoma cells,providing a potential therapeutic target for ameloblastoma.

Objective:To reevaluate the systematic reviews on application effect of high-flow nasal cannula oxygen therapy in preterm infants.Methods:Systematic reviews or Meta-analysis on the use of high-flow nasal cannula oxygen therapy in preterm infants were searched by computer from Cochrane Library, PubMed, Embase, CNKI and Wanfang databases. The retrieval period was from establishment of databases to December 6, 2022. Literature screening and data extraction were conducted independently by two researchers. A Measure Tool to Assess Systematic ReviewsⅡ and The Grading of Recommendations Assessment, Development and Evaluation system were used to evaluate the methodological quality and evidence quality grading of the included literature, respectively.Results:A total of 13 systematic reviews were included, and the results showed that the methodological quality of 4 literatures was low, and that of 9 literatures was very low. The hierarchy of evidence of 112 outcome indicators was evaluated. The results showed that the evidence quality of 7 outcome indicators was extremely low, evidence quality of 78 was low, evidence quality of 26 was intermediate and 1 was high.Conclusions:High-flow nasal cannula oxygen therapy is not sufficient to replace the role of non-invasive positive pressure ventilation in respiratory support for premature infants, but it can reduce the incidence of nasal injury and the incidence of air leakage or pneumothorax in respiratory support after extubation. The methodological quality of systematic evaluation included in the study is low, and the credibility of evidence is insufficient. Clinical application needs to carefully adopt relevant evidence and further carry out more standardized and rigorous research.

Background Trichloroethylene (TCE) can enter human body through biological accumulation of polluted water or air, resulting in health hazards. The most commonly involved organs are the liver. Objective To observe potential polarization of M1 Kupffer cells (KCs) in mice liver exposed to TCE orally, and to investigate the relationship between histones lysin demethylase JMJD3 and M1 KCs polarization. Methods A total of 72 SPF BALB/c mice aged 6 to 8 weeks were randomly divided into a blank control group (n=18), a vehicle control group (n=18), a 2.5 mg·mL⁻¹ TCE group (n=18), and a 5.0 mg·mL⁻¹ TCE group (n=18) after adaptive feed for one week. A TCE transoral exposure model was established after eight weeks of administration according to previous research of the research group. In the 2nd, 4th, and 8th weeks, the mice were sacrificed and liver tissue samples were collected. Western blotting was used to detect the expression level of JMJD3 in the liver tissue samples. Immunofluorescence was used to co-locate the macrophage marker F4/80 and the surface marker CD11c of M1 macrophages. Immunohistochemistry was used to detect the expressions of CD16/32, a marker of M1 macrophages, and TNF-α, an inflammatory factor of M1 macrophages in mouse liver. Results In the 2nd, 4th, and 8th weeks, the mice in each group were generally in good condition, and no individual died due to TCE. There was no statistically significant difference in the amount of water consumed by each group, nor in the body weight gain and the liver coefficient of mice at each time point (P>0.05). The results of Western blotting analysis showed that there was no statistically significant difference in JMJD3 protein expression level between the blank control group and the vehicle control group at each time point, the expression levels of JMJD3 protein in the 2.5 mg·mL⁻¹ TCE group and the 5.0 mg·mL⁻¹ TCE group were higher than that in the control group , and the expression level of JMJD3 protein in the 5.0 mg·mL⁻¹ TCE group was higher than that in the 2.5 mg·mL⁻¹ TCE group (P<0.05). The results of immunofluorescence co-localization showed that the expressions of F4/80 and CD11c were low in the blank control group and the vehicle control group, while the expressions of F4/80 and CD11c were increased in the 2.5 mg·mL⁻¹ and the 5.0 mg·mL⁻¹ TCE groups. The results of immunohistochemistry showed that the expressions of CD16/32 and TNF-α in the blank control group and the vehicle control group were low, and there were large deposits in the 2.5 mg·mL⁻¹ TCE group and the 5.0 mg·mL⁻¹ TCE group. Conclusion The polarization of M1 KCs and the expression of proinflammatory factors may be related to an increased expression level of JMJD3 induced by oral TCE exposure.

Objective: To study the resistance characteristics, carbapenemase genotypes and the homology of carbapenem-resistantEscherichia coli(CREC). Methods: 6 092Escherichia coliisolated from clinical specimens in The First Affiliated Hospital of Anhui Medical University were collected and 71 strains of CREC were selected. The identification and antimicrobial susceptibility test were carried out by Vitek-2 Compact. Confirmation of carbapenemase phenotype was performed by modified hodge test(MHT), modified carbapenem inactivation method(mCIM) and carbapenemase nordmann-poirel(Carba NP) test. Carbapenemase-encoding genes(blaKPC, blaNDM, blaVIM, blaIMP,etc.) were identified by PCR and positive amplification products were sequenced, and then analyzed by using BLAST programs. ERIC-PCR fingerprinting was used to determine the clonal relationship between the different strains. Results: CREC strains were mainly distributed in intensive care unit(ICU) and burn department, and the source of specimens was mainly urine. The drug susceptibility results showed that the resistance rates of CREC to ciprofloxacin, levofloxacin and cotrimoxazole were all above 70%, and the resistance rates to amikacin and tobramycin were less than 50%. Among 71 CREC strains, the number of positive strains for MHT, mCIM and Carba NP were 45, 67 and 69, and the positive rates were 63.38%, 94.37% and 97.18%, respectively. Carbapenemase genes were detected in 43 CREC isolates, of which 34 strains(79.07%, 34/43) carried blaNDM, 9 strains(20.93%, 9/43) carried blaKPC-2. In addition, the rates of strains harbored blaNDM-1or blaNDM-5were 20.59%(7/34) and 79.41%(27/34), respectively. Other carbapenem genes such as blaIMP, blaVIMand blaOXA-48were not detected. According to the fingerprint of ERIC-PCR, CREC was divided into 19 genotypes A-S, and no dominant genotype was found. Conclusion Drug resistance rate of clinically isolated CREC in our hospital is high, showing multi-drug resistance. blaNDMis the main carbapenemase gene of CREC. The epidemic CREC in our hospital has high genetic diversity and the homology of CREC is dispersive.

Objective: To investigate the mechanism of cathepsin L(CTSL)-mediated kidney injury in trichloroethene(TCE)-sensitized mice. Methods: 41 BALB/C mice were randomly divided into blank group(n=5), solvent group(n=5), TCE treatment group(n=15) and TCE+CTSLi treatment group(n=16). TCE percutaneous sensitization mouse model was established, and the mice were evaluated as positive group and negative group according to skin sensitization score. The renal pathology of mice was observed by electron microscopy and HE staining, the renal function level of mice was assessed by serum urea nitrogen(BUN). The expression of CTSL was detected by immunofluorescence, the apoptosis of renal cells was assessed by TUNEL staining, and the activation of renal protein kinase C(PKC) signal molecule was detected by Western blot. Results: The sensitization rates of TCE treatment group and TCE+CTSLi treatment group were 53.3%(8/15) and 50.0%(8/16), respectively, and there was no statistical difference in sensitization rates(P>0.05). Pathological results showed that TCE sensitized-mice showed edema and vacuolar degeneration of renal tubular cells, thickening of glomerular basement membrane, fusion of podocytes and mitochondria vacuolar degeneration. The results of renal function showed that the serum BUN level of TCE sensitized mice was higher than that of other groups. Immunofluorescence results showed that the expression level of CTSL in the kidney of TCE-sensitized positive mice increased(P<0.05,F=82.438), and the apoptosis level of renal structure cells was also higher than that of other groups(P<0.05). Western blot showed that the phosphorylation of PKC protein in the kidney of TCE-sensitized mice increased, while the expression of PKC protein in TCE+CTSLi sensitized mice was down-regulated after CTSLi pretreatment(P<0.05,F=35.686), the level of renal cell apoptosis decreased, and renal damage was improved. Conclusion CTSL might aggravate renal damage via activation of PKC signaling in TCE-sensitized mouse.

Porphyromonas gingivalis peptidylarginine deiminase (PPAD), an isoenzyme of animal endogenous peptidylarginine deaminase, is secreted by the Por system and catalyzes the citrullination of arginine. Recent studies have found that PPAD can affect the formation of Porphyromonas gingivalis biofilm and reduce the body′s immune defense function, which is related to the occurrence and development of many diseases such as periodontal diseases and rheumatoid arthritis. In this paper, we reviewed the molecular characteristics of PPAD, including the genetic and functional characteristics, as well as the mechanisms related to the inflammatory and autoimmune diseases. We also pointed some issues that should be pay attention to in the further study.