Tuberculosis (TB), a chronic infectious disease caused by Mycobacterium tuberculosis, is primarily airborne and remains a global health problem, especially in resource-limited countries and regions. The emergence of drug resistance in M. tuberculosis has rendered the existing means ineffective in the treatment of TB. Therefore, research in new therapeutic directions has become imperative. In this review, we outline functional peptides in terms of the mechanisms of action, anti-TB attempts, advantages and disadvantages, and latest advances, aiming to analyze the research progress in anti-TB peptides. Furthermore, we investigate the potential applications of bioactive compounds found in traditional Chinese Medicine within the context of peptides.

ObjectiveTo explore the mechanism through which Banxia Baizhu Tianmatang ameliorates cognitive impairment in epileptic rats induced by lithium chloride-pilocarpine by regulating the neuroinflammatory reaction mediated by NOD-like receptor protein 3 （NLRP3） inflammasomes. MethodsSixty male SD rats were randomly allocated into blank， model， carbamazepine （0.125 g·kg^-1·d^-1）， Banxia Baizhu Tianmatang （1.04 g·kg^-1·d^-1）， and carbamazepine （0.125 g·kg^-1·d^-1） + Banxia Baizhu Tianmatang （1.04 g·kg^-1·d^-1） groups （n=12）. After the modeling of epilepsy， rats were administrated with corresponding agents by gavage for 12 weeks. At the 6th and 12th week of the intervention， the rats’ hyper-excited behavior was evaluated by the stylus experiment， and at the 12^th week of intervention， the cognitive function was evaluated by Barnes maze. At the same time， the seizure frequency and severity grade （Racine score） were recorded. The serum and hippocampus tissue samples were collected after anesthesia for the following tests. Nissl staining was used to evaluate the degree of neuronal damage in the hippocampal CA1 area. The content of malondialdehyde （MDA） in the hippocampus was determined by the thiobarbituric acid （TBA） method. Serum levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18） were quantified by enzyme-linked immunosorbent assay （ELISA）. Immunohistochemical method was adopted to detect the expression of apoptosis-associated speck-like protein containing a card （ASC） in the hippocampus. Western blot was employed to quantitatively analyze the protein levels of NLRP3， cysteinyl aspartate-specific proteinase-1 （Caspase-1）， and brain-derived neurotrophic factor （BDNF） in the hippocampus. ResultsThe model group showed increased stylus scores at the 6th and 12^th week after modeling， a decreased Barnes maze strategy score at the 12^th week， a prolonged incubation period （P<0.05）， elevated serum levels of inflammatory factors （P<0.05）， decreased neurons with scattered arrangement and large gaps in the hippocampus， increased content of MDA in the hippocampus （P<0.05）， an increased positive expression of ASC， and up-regulated protein levels of Caspase-1， NLRP3， and BDNF （P<0.05）. Compared with the model group， the intervention with Banxia Baizhu Tianmatang for 12 weeks was accompanied by a decreased stylus score， epileptic seizures with a decreased score， a decreased number， and shortened duration， an increased Barnes maze strategy score， shortened escape latency （P<0.01）， declined serum levels of inflammatory factors （P<0.05）， regular morphology of hippocampal neurons， reduced MDA content in the hippocampus （P<0.05）， a decreased positive expression of ASC， and down-regulated protein levels of Caspase-1， NLRP3， and BDNF （P<0.05， P<0.01）. In addition， compared with the carbamazepine group， Banxia Baizhu Tianmatang + carbamazepine showed improved performance in controlling the seizure， improved the cognitive behavior score and morphology of hippocampal neurons， alleviated the oxidative stress products， lowered the levels of inflammatory factors， reduced the positive expression of ASC in the hippocampus， and down-regulated the expression of Caspase-1， NLRP3 and BDNF， with no significant differences. ConclusionBanxia Baizhu Tianmatang may reduce neuroinflammation， control epileptic seizures， and ameliorate cognitive impairment by inhibiting the expression of NLRP3 inflammasomes.

Objective To analyze the target,signal pathway and potential mechanism of Banxia Baizhu Tianma Decoction in the treatment of epilepsy based on network pharmacology,and to verify it by molecular docking technology and animal experiments.Methods The active ingredients and drug targets of Banxia Baizhu Tianma Decoction were screened by BATMAN and other databases.The targets of epilepsy-related diseases were obtained by GeneCards and other databases,and the intersection targets were taken.Constructing'drug-ingredient-target-disease'network and PPI network to screen the core targets.The core active ingredients were screened according to GO,KEGG functional enrichment analysis and'pathway-target-active ingredient'network.Molecular docking was used to verify the core targets and core active ingredients.In the animal experiment,the rat model of epilepsy was induced by lithium chloride-pilocarpine,and 40 Wistar rats were divided into normal control group,model control group,carbamazepine group and Banxia Baizhu Tianma Decoction group.The seizures were observed by behavior.Nissl staining was used to observe neuronal damage in hippocampus.Immunohistochemistry was used to detect the expression of BAX,BCL-2 and Caspase-3 protein.Results A total of 1072 targets of Banxia Baizhu Tianma Decoction were screened,1046 disease targets of epilepsy were screened,and 220 intersection targets of Banxia Baizhu Tianma Decoction in the treatment of epilepsy were screened.The core targets AKT1,ALB,ACTB,INS and TNF of PPI network were obtained.KEGG pathway mainly involves TNF signaling pathway,IL-17 signaling pathway,cAMP signaling pathway,pathways of neurodegeneration-multiple diseases,serotonergic synapse and Dopaminergic synapse.The core active ingredients with the highest correlation in the'pathway-target-active component'network were Betulin,Ephedrine,Ergotamine and Thymol.Results of molecular docking indicated that the core target had satisfactory affinity with those active ingredients.Animal experiments showed that Banxia Baizhu Tianma Decoction could effectively reduce epileptic seizures in rats,improve hippocampal neuronal damage in rats,significantly reduce the percentage of neuronal apoptosis,significantly down-regulate the expression of pro-apoptotic proteins BAX and Caspase-3,and up-regulate the expression of anti-apoptotic protein BCL-2.Conclusion Banxia Baizhu Tianma Decoction has the characteristics of multi-component,multi-target and multi-pathway in the treatment of epilepsy.Inhibiting neuronal apoptosis and reducing hippocampal neuronal damage may be one of the important mechanisms for its treatment of epilepsy.

Objective To analyze the target,signal pathway and potential mechanism of Banxia Baizhu Tianma Decoction in the treatment of epilepsy based on network pharmacology,and to verify it by molecular docking technology and animal experiments.Methods The active ingredients and drug targets of Banxia Baizhu Tianma Decoction were screened by BATMAN and other databases.The targets of epilepsy-related diseases were obtained by GeneCards and other databases,and the intersection targets were taken.Constructing'drug-ingredient-target-disease'network and PPI network to screen the core targets.The core active ingredients were screened according to GO,KEGG functional enrichment analysis and'pathway-target-active ingredient'network.Molecular docking was used to verify the core targets and core active ingredients.In the animal experiment,the rat model of epilepsy was induced by lithium chloride-pilocarpine,and 40 Wistar rats were divided into normal control group,model control group,carbamazepine group and Banxia Baizhu Tianma Decoction group.The seizures were observed by behavior.Nissl staining was used to observe neuronal damage in hippocampus.Immunohistochemistry was used to detect the expression of BAX,BCL-2 and Caspase-3 protein.Results A total of 1072 targets of Banxia Baizhu Tianma Decoction were screened,1046 disease targets of epilepsy were screened,and 220 intersection targets of Banxia Baizhu Tianma Decoction in the treatment of epilepsy were screened.The core targets AKT1,ALB,ACTB,INS and TNF of PPI network were obtained.KEGG pathway mainly involves TNF signaling pathway,IL-17 signaling pathway,cAMP signaling pathway,pathways of neurodegeneration-multiple diseases,serotonergic synapse and Dopaminergic synapse.The core active ingredients with the highest correlation in the'pathway-target-active component'network were Betulin,Ephedrine,Ergotamine and Thymol.Results of molecular docking indicated that the core target had satisfactory affinity with those active ingredients.Animal experiments showed that Banxia Baizhu Tianma Decoction could effectively reduce epileptic seizures in rats,improve hippocampal neuronal damage in rats,significantly reduce the percentage of neuronal apoptosis,significantly down-regulate the expression of pro-apoptotic proteins BAX and Caspase-3,and up-regulate the expression of anti-apoptotic protein BCL-2.Conclusion Banxia Baizhu Tianma Decoction has the characteristics of multi-component,multi-target and multi-pathway in the treatment of epilepsy.Inhibiting neuronal apoptosis and reducing hippocampal neuronal damage may be one of the important mechanisms for its treatment of epilepsy.

Background and purpose:SEC14L1P1,a pseudogene of the SEC14 family,is closely associated with the development of various tumors,but its role in oral squamous cell carcinoma(OSCC)has not been clarified.This study aimed to gain insights into the expression characteristics and subcellular localization of SEC14L1P1 in OSCC cells,as well as its effects on OSCC cell proliferation and migration.Methods:The expression of SEC14L1P1 in head and neck squamous cell carcinoma(HNSCC)tissues was analyzed by the ENCORI database;The expression of SEC14L1P1 and its relationship with patient prognosis in HNSCC was further analyzed using the GDC and UCSC Xena databases.The expression of SEC14L1P1 in OSCC cell lines was detected by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR);RNA nucleoplasmic separation assay was performed to determine the localization of SEC14L1P1 in OSCC cells.SEC14L1P1 knockdown(SS-SEC14L1P1)group and knockdown control(SS-NC)group were established for CAL-27 cells,and SEC14L1P1 overexpression(SEC14L1P1)group and overexpression control(Vector)group were established for HN30 cells.The effects of SEC14L1P1 expression on the proliferation and migration abilities of cells in each group were assessed by cell counting kit-8(CCK-8)and transwell migration assays.RTFQ-PCR and Western blot experiments were used to detect the effects of altered SEC14L1P1 expression on the expression levels of epithelial-mesenchymal transition(EMT)-related genes.To investigate the effects of SEC14L1P1 on the proliferation of OSCC cells in vivo using a subcutaneous xenograft tumor model in nude mice,12 four-week-old BALB/c nude mice were randomly divided into two groups:the antisense oligonucleotide(ASO)-NC group and the ASO-SEC14L1P1 group,with 6 mice in each group.All mice were individually labeled.Further mechanistic studies were performed by analyzing molecules interacting with SEC14L1P1 through the RNAInter database,and the ENCORI database was queried for expression correlation between SEC14L1P1 and DHX9.The effect of altered SEC14L1P1 expression on the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)pathway was detected by Western blot assay.Results:Database analysis showed that the expression of SEC14L1P1 was higher in HNSCC tissues than in normal tissues,and was strongly associated with poor patient prognosis.The RTFQ-PCR results showed that SEC14L1P1 was highly expressed in all six OSCC cell lines;RNA nucleoplasmic separation showed that SEC14L1P1 was mainly localized in the nucleus in CAL-27 and HN30 cells.Compared with SS-NC,the relative expression of SEC14L1P1 in the SS-SEC14L1P1 group was significantly lower and significantly inhibited cell proliferation and migration,while the relative expression of SEC14L1P1 in the SEC14L1P1 group was significantly higher compared with the Vector group,which also significantly increased cell proliferation and migration.The down-regulation of SEC14L1P1 was accompanied by increased mRNA and protein levels of E-cadherin,and decreased mRNA and protein levels of N-cadherin and vimentin,with the opposite result after SEC14L1P1 overexpression.In vivo experiments showed that the xenograft tumor weight and volume of the ASO-SEC14L1P1 group were significantly reduced.Further mechanistic studies revealed a positive correlation between SEC14L1P1 and DHX9 expressions,and DHX9 has been shown to activate the PI3K/AKT signaling pathway.Knockdown of SEC14L1P1 resulted in decreased protein expressions of phosphorylated-PI3K(p-PI3K)and phosphorylated-AKT(p-AKT),and overexpression of SEC14L1P1 increased protein expressions of p-PI3K and p-AKT.Conclusion:SEC14L1P1 showed high expression levels in OSCC cells and tissues and promoted the proliferation and migration of OSCC cells,a phenomenon that may be related to the regulation of the PI3K/AKT signaling pathway by SEC14L1P1,which in turn promotes EMT.

Objective This study aims to investigate the effects of acupuncture on the spatial learning and memory capacities,the morphological characteristics of hippocampal neurons,the components of extracellular matrix,as well as the structure of the blood-brain barrier in SAMP8 mice.Moreover,it endeavors to explore the underlying mechanism through which the"Tongdu Qishen"(activating governor vessel and awakening spirit)acupuncture method ameliorates the cognitive function and modulates cerebral blood flow in SAMP8 mice.Methods Twenty 6-month-old male SAMP8 mice were randomly allocated into a model group and an electroacupuncture group,with ten mice in each group.Additionally,ten homologous 6-month-old male SAMR1 mice were designated as the normal control group.In the electroacupuncture group,the acupoint"Renzhong"(GV 26)was punctured using a quick insertion technique,while"Baihui"(GV 20)and"Yintang"(EX-HN 3)were punctured via a transverse insertion method.Subsequently,the needle handles were connected to an electroacupuncture apparatus,which was set to operate with a dense-sparse wave at a frequency of 100 Hz/2 Hz and an electric current of 2 mA.Each session of acupuncture lasted for 20 minutes and was conducted once a day for a consecutive period of 28 days.The normal group,the model group,and the electroacupuncture group all underwent identical fixation and restraint procedures to ensure consistent experimental conditions.The Morris water maze was employed to evaluate the spatial learning and memory abilities of the mice.The Nissl staining technique was utilized to assess the alterations in hippocampal neurons of SAMP8 mice.The laser speckle imaging system was adopted to measure the cerebral blood flow of the mice in each group.Transmission electron microscopy was applied to observe the ultrastructure of the blood-brain barrier within the frontal cortex tissue of the SAMP8 mice's brains.The Western blotting method was used to detect the expressions of Collagen-Ⅳ and Fibronectin in the brain tissues of SAMP8 mice.Results In comparison with the model group,the"Tongdu Qishen"acupuncture method demonstrated a significant efficacy in improving the spatial learning and memory abilities of SAMP8 mice(P<0.05).It also effectively alleviated the degeneration of hippocampal neurons,remarkably increased the cerebral blood flow perfusion volume of SAMP8 mice(P<0.05),ameliorated the damage to the blood-brain barrier structure,significantly upregulated the content of Collagen-Ⅳ(P<0.05),and significantly downregulated the content of Fibronectin(P<0.0001).Conclusion The"Tongdu Qishen"acupuncture method is capable of favorably regulating the extracellular matrix of SAMP8 mice,thereby improving cerebral blood flow and ultimately enhancing their learning and memory abilities.

Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.

The synthetic PDCoV N protein gene was optimized and cloned into the pET-30a vector to obtain the pET-30a-N plasmid.Thenthe recombinant plasmid was transformed into three strains of BL21 E.coli using heat-shock to explore protein expression conditions.The expressed proteins was purified using Ni Focurose 6FF(IMAC)and used as antigen to immunize the New Zealand White rabbit to prepare the polyclonal antibody against the PDCoV N protein.The antibody titer was measured by indirect ELISA method.The specificity for the antibody was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the pET-30a-N plasmid showed high expression level in BL21 StarTM(DE 3).The optimal expression condition was 37 ℃ 4 h.The purity of the target protein could reach 90.3％after purification.Indirect ELISA showed that the antibody titers was up to 1∶204 800.Western blot and IFA showed that the produced rabbit polyclonal antibody exhibited good specificity.In conclusion,the polyclonal antibody was prepared which specifically recognized the PDCoV N proteins.The results provided some references for the subsequent exploration of PDCoV N protein function and laid a foundation for establishing a diag-nostic method for PDCoV.

Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.

The synthetic PDCoV N protein gene was optimized and cloned into the pET-30a vector to obtain the pET-30a-N plasmid.Thenthe recombinant plasmid was transformed into three strains of BL21 E.coli using heat-shock to explore protein expression conditions.The expressed proteins was purified using Ni Focurose 6FF(IMAC)and used as antigen to immunize the New Zealand White rabbit to prepare the polyclonal antibody against the PDCoV N protein.The antibody titer was measured by indirect ELISA method.The specificity for the antibody was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the pET-30a-N plasmid showed high expression level in BL21 StarTM(DE 3).The optimal expression condition was 37 ℃ 4 h.The purity of the target protein could reach 90.3％after purification.Indirect ELISA showed that the antibody titers was up to 1∶204 800.Western blot and IFA showed that the produced rabbit polyclonal antibody exhibited good specificity.In conclusion,the polyclonal antibody was prepared which specifically recognized the PDCoV N proteins.The results provided some references for the subsequent exploration of PDCoV N protein function and laid a foundation for establishing a diag-nostic method for PDCoV.