Prevalence of Streptococcus suis Isolated from Clinically Healthy Sows in China

Streptococcus suis is an important pathogen in pigs. Transmission of this pathogen is generally believed to occur between healthy carrier sows and their offspring, so the carrier status of S. suis in healthy sows is important for the control of S. suis infections in pigs, especially in suckling and growing pigs. In this study, the prevalence of S. suis isolated from clinically healthy sows in China was studied for the first time. A total of 1 043 tonsil samples were collected from clinically healthy sows from 10 regions in China from 2005 to 2007. Among the 421 S. suis isolates, 31 strains were identified as capsular type 2. The results showed that S. suis was widespread in swine herds in China with the carrier rates in different herds ranging from 19.5 to 93.9%. Overall, 40.4 and 3.0% of clinically healthy sows harbored S. suis and capsular type 2 in their palatine tonsils, respectively. Statistically significant differences of carrier rates of S. suis and capsular type 2 between the different farms were observed, which was independent of herd sizes and geographic distributions of different herds.

Comparative Research on Serogroups Distribution and Antimicrobial Resistance of Escherichia coil Isolates from Poultry in Different Areas of China

A total of 241 Escherichia coli （E. coli） isolates from 349 avian samples （292 from cloacae, 29 from feed and water, 28 from dust and padding） were collected from Northeast, South, North, and Central China in recent years. The percentage of isolation was 69.1%. There are 67 serogroups each with 1-2 isolates distributed in different regions, and some of these regions had the preponderant serogroups. Antimicrobial-resistance （AR） of E. coli was so severe that the majority were multi-AR. Fifty percent strains were resistant to 10-19 antimicrobial drugs. Overall, the isolates represented resistance to nalidixic acid （88.1%）, tetracycline （85.7%）, sulfamethoxazole （81.0%）, trimethoprim-sulfamethpxazole （77.1%）, ampicillin （76.2%）, amoxilline （74.3%）, streplomycin （66.2%）, fluoroquinolones （57.1-66.7%）, chloramphenicol （52.9%）, gentamicin （39.0%）, and kanamycin （36.2%）. The isolates were sensitive to cefalexin, amoxilline-clavulanic acid, amikacin, and florfenicol with an AR rate of 0-19,5% only, The results showed that the AR was more severe in chicken farms in which the antibiotics were used broadly and repeatedly. This study indicated the AR characterization of E, coli in different areas of China. It will be a foundation for studying AR mechanism and regulating the usage of antimicrobial in the poultry industry.

Molecular Epidemiology of Klebsiella pneumoniae from Clinical Bovine Mastitis in Northern Area of China,2018–2019

Klebsiella pneumoniae(K.pneumonia,KpⅠ)is a predominate inducement of bovine mastitis,which is associated with high mortality and milk yield reduction.However,data is lacking on the molecular characteristics of bovine K.pneumoniae,limiting the risk assessment of its transmission through the food chain.Herein,we investigated the prevalence of K.pneumoniae in 6301 clinical mastitis(CM)milk samples from dairy cattle in northern area of China.In total,183 K.pneumoniae isolates were recovered,with detection rates of 3.0% and 2.8% in 2018 and 2019,respectively.Like human clinical K.pneumoniae,all CM K.pneumoniae isolates belonged to one of three phylogroups:KpⅠ(n=143),Klebsiella.quasipneumoniae subsp.similipneumoniae(KpⅡ-B)(n=37),and Klebsiella variicola(KpⅢ)(n=3).We detected the extendedspectrum β-lactamase-encoding genes bla_(SHV-2a),blac_(CTX-M-14),and bla_(CTX-M-15),as well as clpC,lpfA,lacI,lacZ,lacY,and the fecABDEIR operon in the KpⅠ isolates,which may contribute to their pathogenicity and host adaptability in cows.The high prevalence of KpⅠ in dairy farms may be problematic,as it showed relatively higher rates of antibiotic resistance and virulence gene carriage than the KpⅡ-B and KpⅢ isolates.Furthermore,we observed distinct differences in population structure between CM-and human infection-associated KpⅠ isolates,with the genes associated with invasive infection in humans rarely being observed in bovine isolates,indicating that few CM-associated K.pneumoniae isolates pose a threat to human health.Nevertheless,bovine KpⅡ-B isolates shared a high level of nucleotide sequence identity with isolates from human infections and frequently carried the nitrogen-fixation gene nif,suggesting an association between KpⅡ-B isolates from cattle and humans,and plant-derived bacteria.

A multiplex real-time PCR assay for simultaneous detection of classical swine fever virus,African swine fever virus,and atypical porcine pestivirus

With the implementation of the C-strain vaccine,classical swine fever(CSF) has been under control in China,which is currently in a chronic atypical epidemic situation.African swine fever(ASF) emerged in China in 2018 and spread quickly across the country.It is presently occurring sporadically due to the lack of commercial vaccines and farmers’ increased awareness of biosafety.Atypical porcine pestivirus(APPV) was first detected in Guangdong Province,China,in 2016,which mainly harms piglets and has a local epidemic situation in southern China.These three diseases have similar clinical symptoms in pig herds,which cause considerable losses to the pig industry.They are difficult to be distinguished only by clinical diagnosis.Therefore,developing an early and accurate simultaneous detection and differential diagnosis of the diseases induced by these viruses is essential.In this study,three pairs of specific primers and Taq-man probes were designed from highly conserved genomic regions of CSFV(5’ UTR),African swine fever virus(ASFV)(B646L),and APPV(5’ UTR),followed by the optimization of reaction conditions to establish a multiplex real-time PCR detection assay.The results showed that the method did not cross-react with other swine pathogens(porcine circovirus type 2(PCV2),porcine reproductive and respiratory syndrome virus(PRRSV),foot-and-mouth disease virus(FMDV),pseudorabies virus(PRV),porcine parvovirus(PPV),and bovine viral diarrhea virus BVDV).The sensitivity results showed that CSFV,ASFV,and APPV could be detected as low as 1 copy μL–1;the repeatability results showed that the intra-assay and interassay coefficient of variation of ASFV,CSFV,and APPV was less than 1%.Twenty-two virus samples were detected by the multiplex real-time PCR,compared with national standard diagnostic and patented method assay for CSF(GB/T 27540–2011),ASF(GB/T 18648–2020),and APPV(CN108611442A),respectively.The sensitivity of this triple real-time PCR for CSFV,ASFV,and APPV was almost the same,and the compliance results were the same(100%).A total of 451 clinical samples were detected,and the results showed that the positive rates of CSFV,ASFV,and APPV were 0.22% (1/451),1.3%(6/451),and 0%(0/451),respectively.This assay provides a valuale tool for rapid detection and accurate diagnosis of CSFV,ASFV,and APPV.

Sequence and phylogenetic analysis of chicken reoviruses in China

supported by the China Animal Disease Prevention and Control Center;the China Agriculture Research System Poultry-Related Science and Technology Innovation Team of Peking, China （CARS-PSTP）

A Novel RT-LAMP Assay for Rapid and Simple Detection of Classical Swine Fever Virus

A simple and rapid assay for the detection of Classical swine fever virus(CSFV)was established using reverse transcription loop-mediated isothermal amplification(RT-LAMP).This study describes the amplification of the genomic RNA of CSFV under isothermal conditions(63℃)within one hour,using a set of six primers(two outer primers,two inner primers and two loop primers).This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR.This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV.PRRSV.SIV. PRV-PCV,thus showed a good specificity.Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition,either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye.Because RT-LAMP is low-cost and produces rapid results,it has the potential to be an excellent tool for CSFV surveillance in the field,especially in developing countries.

Rapid and Sensitive Detection of PRRSV by a Reverse Transcription-Loop-mediated Isothermal Amplification Assay

A real-time monitoring reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the sensitive and specific detection of prototypic,prevalent North American porcine reproductive and respiratory syndrome virus (PRRSV) strains.As a higher sensitivity and specificity method than reverse transcription polymerase chain reaction (RT-PCR),the RT-LAMP method only used a turbidimeter,exhibited a detection limit corresponding to a 10-4 dilution of template RNA extracted from 250 μL of 105 of the 50% tissue culture infective dose (TCID50) of PRRSV-containing cells,and no cross-reactivity was observed with other related viruses including porcine circovirus type 2,swine influenza virus,porcine rotavirus and classical swine fever virus.From forty-two field samples,33 samples in the RT-LAMP assay was detected positive,whereas three of which were not detected by RT-PCR.Furthermore,in 33 strains of PRRSV,an identical detection rate was observed with the RT-LAMP assay to what were isolated using porcine alveolar macrophages.These findings demonstrated that the RT-LAMP assay has potential clinical applications for the detection of highly pathogenic PRRSV isolates,especially in developing countries.

Development of a Loop-Mediated Isothermal Amplification Assay for Porcine Circovirus Type 2

In this study, the loop-mediated isothermal amplification (LAMP) method was used to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). According to the PCV2 sequences published in GenBank, multiple LAMP primers were designed targeting conserved sequences of PCV2. Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template, LAMP reactions in a PCV2 LAMP system was performed, the amplification products were detected by adding SYBR Green I and could be observed directly by the naked eye. The results showed highly-efficient and specific amplification in 30 min at 63°C with a LAMP real-time turbidimeter. Furthermore, PCV2 DNA templates, with a detection limit of 5.5×10-5 ng of nucleic acid, indicated that this assay was highly sensitive. The results obtained with the naked eye after SYBR Green I staining were consistent with those detected by the real-time turbidimeter, showing the potential simplicity of interpretation of the assay results. The LAMP assay appeared to have greater accuracy than PCR and virus isolation for the analysis of 18 clinical samples. In addition it offers higher specificity and sensitivity, shorter reaction times and simpler procedures than the currently available methods of PCV2 detection. It is therefore a promising tool for the effective and efficient detection of PCV2.

Nested RT-PCR method for the detection of European avian-like H1 swine influenza A virus

Swine influenza A virus（swine IAV） circulates worldwide in pigs and poses a serious public health threat, as evidenced by the 2009 H1N1 influenza pandemic. Among multiple subtypes/lineages of swine influenza A viruses, European avian-like（EA） H1N1 swine IAV has been dominant since 2005 in China and caused infections in humans in 2010. Highly sensitive and specific methods of detection are required to differentiate EA H1N1 swine IAVs from viruses belonging to other lineages and subtypes. In this study, a nested reverse transcription（RT）-PCR assay was developed to detect EA H1 swine IAVs. Two primer sets（outer and inner） were designed specifically to target the viral hemagglutinin genes. Specific PCR products were obtained from all tested EA H1N1 swine IAV isolates, but not from other lineages of H1 swine IAVs, other subtypes of swine IAVs, or other infectious swine viruses. The sensitivity of the nested RT-PCR was improved to 1 plaque forming unit（PFU） m L^（-1） which was over 10~4 PFU m L^（-1） for a previously established multiplex RT-PCR method. The nested RT-PCR results obtained from screening 365 clinical samples were consistent with those obtained using conventional virus isolation methods combined with sequencing. Thus, the nested RT-PCR assay reported herein is more sensitive and suitable for the diagnosis of clinical infections and surveillance of EA H1 swine IAVs in pigs and humans.

Heterogeneity and Secondary Structure Analysis of 3' Untranslated Region in Classical Swine Fever Viruses

The attenuated vaccine strains of CSFV have a 12-nucleotides （nt） insertion in the 3＇-UTR of genome as compared to that of CSFV virulent strains. In this study, we found a distinct heterogeneity in the 3＇-UTR of attenuated Thiverval and HCLV strains. The longest 3＇-UTR of Thiverval strain was 259 base pairs （bp） with a 32-nt insertion, the shortest 3＇-UTR had only 233 bp with a 6-nt insertion. The longest 3＇-UTR of HCLV strain was 244 bp with a 17-nt insertion and the shortest 3＇ UTR was 235 bp with a 8-nt insertion. Compared with the published sequences of 3＇-UTR of vaccine and virulent strains, the 3＇-UTR of CSFV vaccine strains have two variable regions where insertion among the different vaccine strains were frequently found. The first is located between the second conservative TALk codon and the start of T-rich region where we found the variable length insertion in the same vaccine strain Thiveral or HCLV and the second is located between the end of T-rich region and the front of GAA eodon, however, a 4-nt deletion was found in this region in the virulent Shimen strain. These two regions may represent the ＂hot spot＂ for mutation. Modeling the secondary structures of the 3＇-UTR suggests that the T-rich insertion could result in the change of structure and free energy, thus affecting the stability of the 3＇-UTR structure. These findings will help to understand the mechanism of attenuated vaccines and improve vaccine safety, stability, and efficacy.

Insertion site of FLAG on foot-and-mouth disease virus VP1 G-H loop affects immunogenicity of FLAG

The G-H loop of the foot-and-mouth disease virus（FMDV） virion contains certain dominant immunogenic epitopes, as well as an arginine-glycine-aspartic acid（RGD） motif that is recognized by cell surface integrin receptors. Previous experiments indicate that it is critical to maintain virus structural integrity when inserting an exogenous epitope into the surface of an FMDV structural protein. However, it remains to be determined how factors such as different insertion positions affect interactions among the virus, cells and host immune system. In this study, one infectious c DNA clone of the swine FMDV Cathay topotype strain O/CHA/90 was constructed. Then, a FLAG marker（DYKDDDDK） was inserted upstream（–4） or downstream（＋10） of the RGD motif to generate tagged viruses vFLAG-O/CHA/90 or vO/CHA/90-FLAG, investigating the possibility of expressing foreign antigen and effect on its immunogenicity. Compared to the parental virus, both tagged viruses exhibited similar plaque phenotypes, suckling mouse pathogenicity and antigenicity. Additionally, the FLAGtag insertion position did not change the use of integrin-mediated cell entry by the tagged viruses. Interestingly, both tagged vaccines protected pigs against challenge with the parental virus O/CHA/90 and induced immune responses against FMDV in BALB/c mice and pigs, but only vaccination with vFLAG-O/CHA/90 generated anti-FLAG antibodies. Our findings demonstrated that two sites（RGD–4 and RGD＋10） tolerated the insertion of an exogenous gene in the swine FMDV O/CHA/90 strain. However, only RGD–4 was a novel and appropriate inserting site which could tolerate exogenous FLAG. The resultant tagged virus is a promising candidate for FMD vaccine which can be differentiating infected from vaccinated animals（DIVA）.

Preparation of Lung Targeting Microspheres of Gelatin Ceftiofur Alkali

Gelatin ceftiofur alkali microsphere was prepared to observe its characteristics and evaluate preservation conditions. The glutaraldehyde was increased and the carboxylic methyl chitosan was added to improve the microsphere. The experimental results show microspheres have a better morphology surface and fairly regular structure with 4% glutaraldehyde. The average particle size is 15.84 gm and particle size distribution is narrow which shows a good uniformity. Microsphere size was affected by the stirrer speed, dosing ratio and curing degree. The greater drug loaded is, the better microspheres loading is; but with the increase of drug loading rate, the entrapment efficiency increases first and then decreases. The drug release rate of the microsphere is 24.90% in 0.5 h and 84.90% in 48 h, when CMC-GMs with 4% curing agent is 32.03% in 0.5 h and 88.44% in 48 h. So Gms embedding of ceftiofur alkali are better than CMC-GM. The stability tests show that strong light, high temperature, high humidity have a great influence on the microspheres.

Effects of Dihydromyricetin Extracted from Ampelopsis grossedentata on Visceral Organ Indexes and Intestinal Digestive Enzyme Activities of Chickens

To provide a reference basis for reasonable development and utilization of Ampelopsis grossedentata resource and its application in production,we added 0.025%,0.05% and 0.1% dihydromyricetin（DMY）extracted from A.grossedentata into the basal diets of Yaoshan chickens,and studied the effects of DMY on visceral organ indexes and digestive enzyme activities of 40-day-old healthy Yaoshan chickens.Supplementation of DMY in basal diets influenced visceral organ weight,organ index,length and digestive enzyme activity of small intestine of chickens.Among them,0.05% DMY was the most appropriate volume for supplementation.Compared with control group（without DMY）,0.05% DMY reduced lung weight and index（P〉0.05）,significantly reduced liver weight（P〈0.05）,extremely reduced stomach weight and stomach index（P〈0.01）,increased activity of amylase in small intestine（P〉0.05）,significantly increased length of small intestine and activity of protease（P〈0.05）.Supplementation of 0.05% DMY reduced the visceral organ weight and organ index,enhanced digestion and absorption ability of gastrointestinal tract,thereby improving feed utilization rate and promoting the growth of chickens.

Influence of PPV,PRV and PRRSV on Efficacy of the Lapinized Hog Cholera Vaccine and Pathogenicity of Classical swine fever virus

Classical swine fever caused by Classical swine fever virus （CSFV） is a serious problem for swine industries in developing countries, which successful control of the disease have been relying on vaccination. However, classical swine fever still occurs in some immunized swine herds for various reasons. In this study, we conducted animal experiments to examine the influence of single or mixed infection with Porcine parvo virus （PPV）, Pseudorabies virus （PRV） and Porcine reproductive and respiratory syndrome virus （PRRSV） on the protective immunity induced by the Lapinized hog cholera virus （HCLV） vaccine and the pathogenicity of CSFV. In experiment 1, pigs were first inoculated with PPV, PRV or PRRSV, then immunized with HCLV, and finally challenged with a highly virulent CSFV Shimen strain. All of the pigs immunized with HCLV survived after the challenge, while all of the pigs in the non-immunized control group died after the challenge. The pigs in the group immunized with HCLV did not show any clinical symptoms of classical swine fever and were negative with CSFV after the challenge. The pigs infected with the non-CSFV before HCLV immunization did not display any clinical symptoms after the challenge with CSFV Shiman strain, but 11 of the 12 pigs were positive with CSFV. In experiment 2, pre-infections with PPV, PRV, and PRRSV were followed by inoculation with a low-virulence CSFV strain （CSFV 39）, and then the pigs were challenged with the CSFV Shimen strain. Infections by either PPV, PRV or PRRSV did not enhance the virulence of CSFV-39, but pigs infected by a mixture of the 3 viruses developed clinical symptoms after inoculation with CSFV-39. The mixed infection also increased mortality caused by the challenge with the CSFV Shimen strain. Together, these results showed PPV, PRV and PRRSV infections in pigs can reduce the efficacy of the HCLV vaccine and enhance the pathogenicity of CSFV, which may partly explain the immunization failure against CSFV in some swine herds.

Studies on Antimicrobial Resistance Transfer In vitro and Existent Selectivity of Avian Antimicrobial-Resistant Enterobacteriaccae In vivo

Increasing antimicrobial resistance （AR） has become a severe problem of public health in the world, whereas control of the AR of bacteria will be based on investigation of the AR mechanism. Furthermore, understanding the existent selectivity of AR organisms from animals can prevent the emergence and diffusion of AR effectively. PCR amplifications of gyrA and parC genes have been performed for detecting fluoroquinolones-resistance （FR） genes. A conjugational transfer test has been carried out using a donor which is resistant to tetracycline （TE）, ampicillin （AMP）, sulfamethoxazole-trimethoprim （SXT）, and a recipient which is sensitive to TE, AMP, and SXT. The AR strains have been passed 20 passages. Two groups of chicken inoculated multi-AR Escherichia coli （E. coli） and multi-AR Salmonella, respectively, are mix-fed. The result shows that amino acid codons of Ser-83 and Asp-87 are mutations from gyrA and there are no mutations from parC genes in all the FR strains. Resistance to TE, AM, and SXT can transfer among E. coli and the conjugal transfer frequency of TE is 3×10^-7. AR can inherit in 20 passages at least. The multi-AR E. coli and Salmonella can be isolated from all chickens three days after inoculation but CIP-resistant strains decrease during the time run out and disappear at 23 days after inoculation. The results indicate that the mutations of gene gyrA are correlative with the FR phenotype. AR genes that are not connected to the chromosome can transfer horizontally and vertically. AR bacteria can diffuse quickly and eliminate naturally from the host if the chicken is not under the pressure of this antibiotic.

Integration and development of veterinary Chinese medicine insight into the development of human Chinese medicine

Chinese veterinarians shine brightly in the long river of history.To examine the beginnings and growth of Chinese Medicine(CM)and Chinese Veterinary Medicine(CVM),the evolution of veterinary Chinese medicine and the common use of herbs by humans and animals were presented.This study summarizes the history of development and the current situation of Chinese veterinary medicine.Explain that veterinary Chinese medicine refers to drugs and their preparations applied under the guidance of CVM theory.Focus on distinguishing the similarities and differences between veterinary Chinese medicine and plant medicine,veterinary Chinese medicine with the complexity and efficacy of integrity,low harmful residue,not easy to produce resistance,and other characteristics.Veterinary Chinese medicine places great emphasis on clinical practice.veterinary Chinese medicine has a long history of veterinary clinical experience and data,which reflects the practical characteristics of veterinary Chinese medicine.The application experience of CM in the prevention and control of animal diseases and promoting animal growth was introduced.

Exosomal miR-218 regulates the development of endometritis in dairy cows by targeting TGIF2/TGF-βpathway

Endometritis affects the reproductive capacity of dairy cows and leads to serious economic losses in dairy farming.Clarification of the pathogenesis of endometritis is necessary to improve the reproductive efficiency of dairy cows.Exosomes and their miRNAs have been proven to play an important role in inflammatory regulation.Exosomal miR-218 is a differentially expressed miRNA found in endometrial epithelial cells(EECs)under endometrial inflammation.Therefore,we investigated the expression of miR-218 in the uterine tissue of dairy cows,lipopolysaccharide(LPS)treated EECs,exosomal vesicles,and regulation of exosomal miR-218 by targeting TGIF-2 inducible factor homology frame 2(TGIF2)/transforming growth factor-beta(TGF-β).The expression of miR-218 was suppressed in inflammatory uterine tissues and LPS treated EECs.The expression of TGIF2 and TGF-βin inflammatory uterine tissues and LPS treated EECs was significantly higher than those in healthy uterine tissues and EECs(p<0.01).Interestingly,miR-218 derived from donor cells was found to regulate the expression of the target gene TGIF2 in recipient cells through the fusion of exosomes.Concurrently,the expression of its target gene TGIF2 was also suppressed by miR-218 in donor cells resulting in fewer TGIF2 being transported into recipient cells with exosomal fusion.This may be a novel mechanism of miRNAs-mediated regulation and provides a new reference for analyzing the pathogenesis of endometritis in dairy cows.

Effect of Chanfukang(Chinese Medicine)on Endothelin and Nitric Oxide in Postpartum Cows with Qi Deficiency and Blood Stasis

[ Objective] To observe effect of Chanfukang on vascular endothelial cells of postpartum cows with Oi deficiency and Blood stasis, and to explore the treatment mechanism of Chanfukang. [Method] A total of 58 cows were assigned to four groups. The cows in group 1 and group 2 suffered from Qi deficiency and Blood stasis. The cows in group 1 were gavaged with Chanfukang （0.5 g/kg-BW, 5 d） at 12 h post partum, but the cows in group 2 were not treated. The cows in group 3 were healthy and gavaged with Chanfukang （0.4 g/kg.BW, 5 d） at 12 h post partum. Cows in group 4 were healthy control cows. The plasma endothelin （ET） concentration and serum nitric oxide （NO） concentration were determined before and after parturition, respectively. [ Result ] The plasma ET concentration and serum NO concentration were significantly higher in the cows with Qi deficiency and Blood stasis than in the healthy control cows （P 〈 0.05）. The NO concentration decreased on Day 10 post partum in the cows with Qi deficiency and Blood stasis, but it was still higher than that in the healthy control cows. After taking Chanfukang for 7 d, the ET concentration and NO concentration decreased, and no significant difference was found between the cows with Qi deficiency and Blood stasis treated with Chanfukang and the healthy control cows. [ Conclusion] Chanfukang may relieve Qi deficiency and Blood stasis of postpartum cows through decreasing plasma ET concentration and serum NO concentration.

Geological Settings and Significance of Zigong Global Geopark

This paper introduced fossil resources,geographical settings and significance of Zigong Global Geopark.In addition,in the long history of local well salt production,remarkable ten great science and technology inventions were created,and numerous well salt production relics were perfectly preserved,thus they have high scientific value and significance.On the basis of field survey and detailed researches,the authors analyzed geological settings in the geopark and significance in depth.The results deduced by the authors can help understand the geoheritage resources in Zigong Global Geopark,and also provide a basis for the protection,development and utilization of geoheritage as well as the planning and management of the geopark.

Mori fructus aqueous extracts attenuates liver injury by inhibiting ferroptosis via the Nrf2 pathway

Background Liver fibrosis and hepatocellular carcinogenesis secondary to liver fibrosis are serious liver diseases with no effective treatments.Mori fructus aqueous extracts(MFAEs)have served as successful treatments for many types of liver injury including fibrosis although the molecular mechanisms are unknown at present.Purpose To investigate the effect of MFAEs in alleviating acute and chronic liver injury and tried to decipher the underlying mechanism.Methods and results Mice were divided into 5 groups(n ps:contro=8)for acute(groups:control,0.3%CCl_(4),bifendate(BD),100 and 200 mg/kg MFAEs,7 d)and chronic(groul,10%CCl_(4),BD,100 and 200 mg/kg MFAEs,4 weeks)liver injury study.Each mouse was injected intraperitoneally with 10μL/g corn oil containing CCl_(4)expect the control group.HepG2 cells were used in vitro study.Eighteen communal components were identified by UPLC-LTQOrbitrap-MS.We utilized a mouse model for acute and chronic liver injury using CCl_(4)and MFAEs administration effectively blocked fibrosis and significantly inhibited inflammation in the liver.MFAEs activated the nuclear factor erythroid derived 2 like 2/heme oxygenase 1(Nrf2/HO-1)pathway and promoted the synthesis of the antioxidants glutathione(GSH),superoxidedismutase(SOD)and glutathione peroxidase(GSH-Px)that resulted in reduced levels of CCl_(4)-induced oxidative stress molecules including reactive oxygen species.These extracts administered to mice also inhibited ferroptosis in the liver by regulating the expression of Acyl-CoA synthetase long chain family member 4(ACSL4),solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4),thus reducing the occurrence of liver fibrosis.Both in vivo and in vitro tests indicated that the mechanism of MFAEs protection against liver fibrosis was linked to activation of Nrf2 signaling.These effects were blocked in vitro by the addition of a specific Nrf2 inhibitor.Conclusion MFAEs inhibited oxidative stress,ferroptosis and inflammation of the liver by activating Nrf2 signal pathway and provided a significant protective effect against CCl_(4)-induced liver fibrosis.