Currently,three predominant subtypes of influenza virus are prevalent in pig populations worldwide:H1N1,H3N2,and H1N2.European avian-like H1N1 viruses,which were initially detected in European pig populations in 1979,have been circulating in pigs in eastern China since 2007.In this study,six influenza A viruses were isolated from 60 swine lung samples collected from January to April 2011 in eastern China.Based on whole genome sequencing,molecular characteristics of two isolates were determined.Phylogenetic analysis showed the eight genes of the two isolates were closely related to those of the avian-like H1N1 viruses circulating in pig populations,especially similar to those found in China.Four potential glycosylation sites were observed at positions 13,26,198,277 in the HA1 proteins of the two isolates.Due to the presence of a stop codon at codon 12,the isolates contained truncated PB1-F2 proteins.In this study,the isolates contained 591Q,627E and 701N in the polymerase subunit PB2,which had been shown to be determinants of virulence and host adaptation.The isolates also had a D rather than E at position 92 of the NS1,a marker of mammalian adaptation.Both isolates contained the GPKV motif at the PDZ ligand domain of the 3' end of the NS1,a characteristic marker of the European avian-like swine viruses since about 1999,which is distinct from those of avian,human and classical swine viruses.The M2 proteins of the isolates have the mutation (S31N),a characteristic marker of the European avian-like swine viruses since about 1987,which may confer resistance to amantadine and rimantadine antivirals.Our findings further emphasize the importance of surveillance on the genetic diversity of influenza A viruses in pigs,and raise more concerns about the occurrence of cross-species transmission events.

Objective To investigate the prevalence and molecular features of Cryptosporidium in sheep and goats from Anhui Province and neighboring provinces. Methods A total 832 and 781 fresh fecal samples were collected from seven large-scale sheep farms and ten large-scale goat farms in Anhui Province and neighboring provinces of Henan, Jiangsu and Shandong. The prevalence and species of Cryptosporidium were investigated in the fecal samples from the sheep and goats in the study areas using nested PCR assay based on the Cryptosporidium-specific SSU rDNA gene, and the subgenotypes of C. parvum and C. ubiquitum were characterized by amplification and sequencing of the 60 kDa glycoprotein (gp60) gene. Results The overall prevalence of Cryptosporidium was 5.8% (48/832) in sheep and 8.7% (68/781) in goats in Anhui Province and neighboring provinces, respectively. The SSU rDNA gene-based PCR assay identified C. xiaoi and C. ubiquitum in sheep and C. parvum in goats, and subtyping revealed that all C. ubiquitum subgenotypes belonged to XIIa subtype 2 and C. parvum subgenotypes belonged to IIdA19G1. Conclusion The identification of zoonotic C. ubiquitum XIIa subtype 2 and C. parvum subtype IIdA19G1 suggests that sheep and goats may serve as a potential source for human Cryptosporidium infections.

To investigate the effects of geniposidic acid( GPA) on hepato-enteric circulation in cholestasis rats,and to explore the mechanism based on the sirtuin 1( Sirt1)-farnesol X receptor( FXR) pathway,sixty SD rats were randomly divided into 6 groups:blank control group,ANIT model group,ursodeoxycholic acid group( 100 mg·kg~(-1)·d-1 UDCA),and GPA high,medium and low( 100,50 and 25 mg·kg~(-1)·d-1) dosage groups,10 rats in each group. Corresponding drugs were intragastrically( ig) administered for10 days. After administration on day 8,all rats except blank rats were administered with 65 mg·kg~(-1)α-naphthalene isothiocyanate( ANIT) once. After the last administration,the serum levels of alanine aminotransferase( ALT),glutamine oxalacetate aminotransferase( AST),gamma-glutamyltransferase( γ-GGT),alkaline phosphatase( ALP),total bilirubin( TB) and total bile acid( TBA)were measured,and the mRNA transcription levels of Sirt1,FXR,multidrug resistant associated protein 2( MRP2),bile salt export pump( BSEP),sodium taurocholate contractible polypeptide( NTCP) in liver and apical sodium bile acid transporter( ASBT),ileum bile acid binding protein( IBABP) in ileum were detected by reverse transcription-polymerase chain reaction( RT-PCR). The protein expression levels of Sirt1,FXR and NTCP were detected by Western blot; the expression of MRP2,BSEP in liver and ASBT,IBABP in ileum were determined by immunofluorescence three staining. Primary rat hepatocytes were cultured in vitro to investigate the inhibitory effect of GPA on a potent and selective Sirt1 inhibitor( EX 527),and the mRNA and protein expression levels of Sirt1 and FXR were detected by RT-PCR and Western blot. GPA significantly decreased the levels of ALT,AST,γ-GGT,ALP,TB,TBA in serum( P<0.01) and improved the pathological damage of liver tissues in ANIT-induced cholestasis rats; significantly increased the mRNA and protein expression levels of Sirt1,FXR,MRP2,BSEP,NTCP in liver and ASBT,IBABP in ileum( P< 0.01). In vitro primary hepatocytes experiment indicated that the gene and protein expression levels of FXR and Sirt1 were noticeably improved by GPA in primary hepatocytes inhibited by EX-527( P<0.01). It was found that the improvement of GPA was in a dose-dependent manner. GPA could improve bile acid hepatointestinal circulation and play a liver protection and cholagogu role in cholestasis rats induced by ANIT.The mechanism may be that GPA activated FXR by regulating Sirt1,a key regulator of oxidative stress injury,and then the activated FXR could regulate protein of bile acid hepato-enteric circulation.
Animals ; Cholestasis ; Iridoid Glucosides ; Liver ; Rats ; Rats, Sprague-Dawley ; Receptors, Cytoplasmic and Nuclear ; Signal Transduction ; Sirtuin 1

To screen the active ingredients of Gardenia jasminoides and potential targets,and investigate the mechanisms against cholestasis based on network pharmacology technology. Twenty-one active components of G. jasminoides were retrieved and the target sites were screened by using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform( TCMSP). Cytoscape3. 2. 1 was used to construct the component-target network. Two hundred and eight targets related to cholestasis were searched and screened through Dis Ge NET,KEGG and OMIM databases. The key targets of G. jasminoides components and cholestasis were integrated and screened,and the component-target-disease network was constructed with Cytoscape 3. 2. 1 software to screen out the core network whose freedom degree was greater than the average value. The Clue GO plug-in of Cytoscape 3. 2. 1 software was used to analyze the biological processes and pathway enrichment of G. jasminoides in regulation of cholestasis. GO biological process analysis revealed 17 biological processes,involving 3 signaling biological processes related to cholestasis,i.e. acute inflammatory response,positive regulation of reactive oxygen species metabolic process,and nitric oxide biosynthetic process. KEGG-KEEG-305 terms and REACTOME pathways analysis revealed 17 regulatory pathways,involving 4 signaling pathways related to cholestasis,i.e. metabolism of xenobiotics by cytochrome P450,nuclear receptor transcription pathway,GPVI-mediated activation cascade and platelet activation. It was found that aqueous extract of G. jasminoides could improve serum biochemical abnormalities in ANIT-induced cholestasis rats. Aqueous extract of G. jasminoides could decrease the protein and mRNA expression levels of ESR1 in liver tissues,and increase the protein and mRNA expression levels of PPARG,NOS2,F2 R,NOS3,and NR3 C1. To sum up,the possible mechanisms of G. jasminoides against cholestasis may be related with the above three processes and four pathways.
Animals ; Cholestasis ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Gardenia ; chemistry ; Medicine, Chinese Traditional ; Plant Extracts ; pharmacology ; Rats ; Signal Transduction

OBJECTIVE: To elucidate the action mechanism of Xingnaojing Injection (, XNJI) for sepsis, and to target screen the potential bioactive ingredients. METHODS: An integrated protocol that combines in silico target screen (molecular docking) and database mapping was employed to find the potential inhibitors from XNJI for the sepsis-related targets and to establish the compound-target (C-T) interaction network. The XNJI's bioactive components database was investigated and the sepsis-associated targets were comprehensively constructed; the 3D structure of adenosine receptor A2a and 5-lipoxygenase proteins were established and evaluated with homology modeling method; system network pharmacology for sepsis treatment was studied between the bioactive ingredients and the sepsis targets using computational biology methods to distinguish inhibitors from non inhibitors for the selected sepsis-related targets and C-T network construction. RESULTS: Multiple bioactive compounds in the XNJI were found to interact with multiple sepsis targets. The 32 bioactive ingredients were generated from XNJI in pharmacological system, and 21 potential targets were predicted to the sepsis disease; the biological activities for some potential inhibitors had been experimentally confirmed, highlighting the reliability of in silico target screen. Further integrated C-T network showed that these bioactive components together probably display synergistic action for sepsis treatment. CONCLUSIONS The uncovered mechanism may offer a superior insight for understanding the theory of the Chinese herbal medicine for combating sepsis. Moreover, the potential inhibitors for the sepsis-related targets may provide a good source to find new lead compounds against sepsis disease.
Arachidonate 5-Lipoxygenase ; metabolism ; Computer Simulation ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Humans ; Injections ; Phytochemicals ; therapeutic use ; Receptor, Adenosine A2A ; metabolism ; Reproducibility of Results ; Sepsis ; drug therapy ; metabolism

OBJECTIVETo speed up seedling production of pasqueflower (Puzlsatilla chinenses) and their modernization in pasqueflower.

METHODWith tissue culture method, primary culture of different explants, culture of cluster buds and their rooting culture were conducted on medium of treatment combinations of adding different hormones.

RESULTThe appropriate medium for different culture stages were MS + 6-BA 1.0-3.0 mg x L(-1) + NAA 0-0.05 mg x L(-1) + Sucrose 30 g x L(-1) in primary culture, MS + 6-BA 0.2 mg x L(-1) + NAA 0.02 mg x L(-1) + BR 0.00001 mg x L(-1) + Sucrose 30 g x L(-1) in differentiation and subculture of cluster buds, 1/2 MS + NAA 0.4 mg x L(-1) + Sucrose 20 g x L(-1) in rooting.

CONCLUSIONApplying stem tip and flower buds as explants, high frequency propagation of seedlings can be achieved with plant tissue culture in Pasqueflower.

Flowers ; growth & development ; Plant Growth Regulators ; pharmacology ; Plant Roots ; growth & development ; Plant Stems ; growth & development ; Plants, Medicinal ; growth & development ; Pulsatilla ; growth & development ; Seedlings ; growth & development ; Tissue Culture Techniques ; methods

The present study was designed to target fish for potential bioactive components contained in a Huang Lian Jie Du decoction (HLJDD) and identify the underlying mechanisms of action for the treatment of sepsis at the molecular level. he bioactive components database of HLJDD was constructed and the sepsis-associated targets were comprehensively investigated. The 3D structures of the PAFR and TXA2R proteins were established using the homology modelling (HM) method, and the molecular effects for sepsis treatment were analysed by comparing the bioactive components database and the sepsis targets using computational biology methods. The results of the screening were validated with biological testing against the human oral epidermal carcinoma cell line KB in vitro. We found that multiple bioactive compounds contained in the HLJDD interacted with multiple targets. We also predicted the promising compound leads for sepsis treatment, and the first 28 compounds were characterized. Several compounds, such as berberine, berberrubine and epiberberine, dose-dependently inhibited PGE2 production in human KB cells, and the effects were similar in the presence or absence of TPA. This study demonstrates a novel approach to identifying natural chemical compounds as new leads for the treatment of sepsis.
Anti-Inflammatory Agents, Non-Steroidal ; pharmacokinetics ; Berberine ; analogs & derivatives ; pharmacokinetics ; Dinoprostone ; biosynthesis ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Humans ; KB Cells ; Platelet Membrane Glycoproteins ; drug effects ; Protein Transport ; Receptors, G-Protein-Coupled ; drug effects ; Receptors, Thromboxane A2, Prostaglandin H2 ; drug effects ; Sepsis ; drug therapy ; metabolism ; Tetradecanoylphorbol Acetate ; pharmacokinetics

Objective:To study the learning curve in laparoscopic left lateral hepatic sectionectomy.Methods:The clinical data of 62 consecutive patients who underwent left lateral hepatic sectionectomy by a single operator from February 2015 to May 2022 in General Hospital of Northern Theater Command were retrospectively analyzed. There were 22 males and 40 females, with mean ±s.d. of (50.7±11.7) years. The learning curve was depicted and evaluated by using the cumulative summation test. The general information, operation and postoperative indicators of the growth level group and the master level group were compared.Results:The average operation time of the 62 consecutive subjects was (172.9±70.1) minutes. Intraoperative blood loss was 100 (50, 200) ml. Two patients were converted to open hepatectomy. Clavien-Dindo grade I postoperative complications occurred in 20 patients (32.3%), with grade Ⅱ in 1 patient (1.6%) and grade Ⅲb in another patient (1.6%). The learning curve reached its highest point on the 20th patient by using the cumulative summation test. The study subjects were then assigned into the growth level group (patient 1-20) and the master level group (patient 21-62). The master level group had a significantly wider spread of patient age [(52.9±11.0) years vs (46.1±11.9) years], decreased operation time [(146.8±55.6) min vs (227.9±66.7) min], shortened drainage tube removal time [4(3, 5) d vs 6(4, 7) d] and decreased postoperative hospital stay [5(5, 7) d vs 6.5(4, 9) d] (all P<0.05) when compared with the growth level group. Conclusion:Left lateral hepatic sectionectomy was safe and feasible, and a single operator went through a learning curve of 20 patients before he/she could master the operation more proficiently.

OBJECTIVETo investigate the effect of clopidogrel on the binding rate of ginsenosides with rat serum proteins (RSA).

METHODSEquilibrium dialysis and liquid chromatography-mass spectrometry were employed to quantify the concentration of ginsenoside Rg1 and Rb1. The protein-binding rates of Rg1 and Rb1 in the presence or absence of clopidogrel (1.0 mg/L) were determined. A molecular simulation model (consisting of homology modeling and molecular docking interaction) was used to reveal the target protein-compound interactions.

RESULTSThe binding rates of ginsenosides Rg1 (0.4, 1.0, and 2.0 mg/L) with RSA were (30.16∓2.82)%, (33.42∓4.21)%, and (34.61∓3.42)%, and those of and Rb1 were (50.13∓2.34)%, (51.23∓3.23)%, and (53.11∓3.26)%, respectively. In the presence of clopidogrel, the binding rates of Rg1 decreased to (22.13∓2.72)%, (21.42∓3.22)%, and (25.45∓3.52)%, and those of Rb1 to (40.13∓3.24)%, (41.25∓4.15)%, and (43.11∓3.31)%, receptively. The molecular docking suggested that these compounds competed to bind with RSA.

CONCLUSIONClopidogrel can competitively bind to RSA with ginsenosides to lower the plasma protein binding rates of ginsenosides.

Objective To evaluate the application and effect of signature verification technology in children's vaccination clinics (CVC) of Jiangsu Province in 2020. Methods The signature verification data were derived from the Jiangsu Provincial Vaccination Integrated Service Management Information System, and the inquiry and registration, informed consent, vaccine traceability code scanning and observation information of children's vaccination clinics in different regions were analyzed. 210 doses of vaccination information were randomly selected from CVCs in each county, and the length of vaccination services in different regions was compared. Results During 2020, all of CVCs in Jiangsu were equipped with signature verification technology, and the signature verification rate of each vaccination sector was more than 99.90%. The length of outpatient vaccination service and overall length of stay in southern Jiangsu were slightly shorter than those in other regions. Conclusion The introduction of electronic signature verification technology in CVCs can effectively standardize the vaccination. It is necessary to expand the functions of electronic signature verification equipment, strengthen data analysis and utilization, and guide vaccination scientifically.