AIM:To investigate the effects of lactylation at the K197 site of peroxiredoxin 1(PRDX1)on the proliferation and migration of glioblastoma cells.METHODS:(1)Immunofluorescence and lactylation pan-antibody techniques were adopted to compare the differences in PRDX1 lactylation modification level between glioblastoma tissues and adjacent normal tissues.High-throughput mass spectrometry and modificomics analysis were utilized to select PRDX1 protein and its K197 site as the focus.(2)Cell experiments were conducted using lactate(5,10 and 15 mmol/L),glu-cose(5,10 and 25 mmol/L)and glycolysis inhibitor 2-deoxy-D-glucose(2-DG;1,5,10 and 15 mmol/L)to treat human glioblastoma U87MG and LN229 cells.Cell proliferation was detected by EdU proliferation staining,and PRDX1 expres-sion was detected in U87MG,LN229 and glial cells via immunoprecipitation and Western blot.The PRDX1 expression and lactylation levels were further examined in 10 mmol/L lactic acid-treated and untreated cells using immunoprecipita-tion and Western blot.(3)The U87MG and LN229 cells were transfected with constructed lactate dehydrogenase A(LDHA)siRNA(si-LDHA)plasmids and negative control(si-Con)plasmids,and the lactylation level of PRDX1 was as-sessed by immunoprecipitation and Western blot.(4)Similarly,the U87MG and LN229 cells were transfected with PRDX1 shRNA(sh-PRDX1)plasmids and negative control(sh-Con)plasmids,and PRDX1 expression was determined by Western blot.(5)The PRDX1 K197R mutant and PRDX1 wild-type(WT)plasmids were constructed and transfected into U87MG and LN229 cells.The PRDX1 expression and lactylation levels were determined by immunoprecipitation and Western blot.The CCK-8 and EdU assays were used to measure cell viability and proliferation,and Transwell assay was performed to assess the migration of U87MG and LN229 cells transfected with PRDX1 K197R mutant and PRDX1 WT plasmids.(6)A tumor formation model in nude mice was established.The LN229 cells with or without PRDX1 K197R mutation were used in the tumor formation experiment with 6 nude mice per group.After 18 d,the nude mice were eutha-nized,and tumor tissues were harvested.Histological changes were observed by HE staining,the lactylation modification leve was detected by immunofluorescence,and immunohistochemistry method was adopted for checking Ki67,a prolifera-tion marker,in tumor tissues.RESULTS:The PRDX1 level in glioblastoma tissues was significantly higher than that in adjacent tissues(P＜0.05).In cell experiments,the addition of lactate and glucose significantly promoted the proliferation and migration of glioblastoma cells and increased the lactylation level of PRDX1(P＜0.05).In contrast,the glycolysis in-hibitor 2-DG inhibited these effects.The si-LDHA transfection experiment showed that knockdown of LDHA reduced the lactylation level of PRDX1(P＜0.05).Importantly,the K197R point mutation in PRDX1 significantly decreased the lacty-lation level of PRDX1 and inhibited the proliferation and migration of glioblastoma cells(P＜0.05).Nude mouse tumori-genesis experiments further confirmed that tumor growth in PRDX1 K197R group was significantly reduced,and the Ki67 proliferation index and lactylation level were decreased(P＜0.05).CONCLUSION:Lactoylation at the K197 site of PRDX1 promotes the proliferation and migration of glioblastoma cells.

AIM:To investigate the effects of lactylation at the K197 site of peroxiredoxin 1(PRDX1)on the proliferation and migration of glioblastoma cells.METHODS:(1)Immunofluorescence and lactylation pan-antibody techniques were adopted to compare the differences in PRDX1 lactylation modification level between glioblastoma tissues and adjacent normal tissues.High-throughput mass spectrometry and modificomics analysis were utilized to select PRDX1 protein and its K197 site as the focus.(2)Cell experiments were conducted using lactate(5,10 and 15 mmol/L),glu-cose(5,10 and 25 mmol/L)and glycolysis inhibitor 2-deoxy-D-glucose(2-DG;1,5,10 and 15 mmol/L)to treat human glioblastoma U87MG and LN229 cells.Cell proliferation was detected by EdU proliferation staining,and PRDX1 expres-sion was detected in U87MG,LN229 and glial cells via immunoprecipitation and Western blot.The PRDX1 expression and lactylation levels were further examined in 10 mmol/L lactic acid-treated and untreated cells using immunoprecipita-tion and Western blot.(3)The U87MG and LN229 cells were transfected with constructed lactate dehydrogenase A(LDHA)siRNA(si-LDHA)plasmids and negative control(si-Con)plasmids,and the lactylation level of PRDX1 was as-sessed by immunoprecipitation and Western blot.(4)Similarly,the U87MG and LN229 cells were transfected with PRDX1 shRNA(sh-PRDX1)plasmids and negative control(sh-Con)plasmids,and PRDX1 expression was determined by Western blot.(5)The PRDX1 K197R mutant and PRDX1 wild-type(WT)plasmids were constructed and transfected into U87MG and LN229 cells.The PRDX1 expression and lactylation levels were determined by immunoprecipitation and Western blot.The CCK-8 and EdU assays were used to measure cell viability and proliferation,and Transwell assay was performed to assess the migration of U87MG and LN229 cells transfected with PRDX1 K197R mutant and PRDX1 WT plasmids.(6)A tumor formation model in nude mice was established.The LN229 cells with or without PRDX1 K197R mutation were used in the tumor formation experiment with 6 nude mice per group.After 18 d,the nude mice were eutha-nized,and tumor tissues were harvested.Histological changes were observed by HE staining,the lactylation modification leve was detected by immunofluorescence,and immunohistochemistry method was adopted for checking Ki67,a prolifera-tion marker,in tumor tissues.RESULTS:The PRDX1 level in glioblastoma tissues was significantly higher than that in adjacent tissues(P＜0.05).In cell experiments,the addition of lactate and glucose significantly promoted the proliferation and migration of glioblastoma cells and increased the lactylation level of PRDX1(P＜0.05).In contrast,the glycolysis in-hibitor 2-DG inhibited these effects.The si-LDHA transfection experiment showed that knockdown of LDHA reduced the lactylation level of PRDX1(P＜0.05).Importantly,the K197R point mutation in PRDX1 significantly decreased the lacty-lation level of PRDX1 and inhibited the proliferation and migration of glioblastoma cells(P＜0.05).Nude mouse tumori-genesis experiments further confirmed that tumor growth in PRDX1 K197R group was significantly reduced,and the Ki67 proliferation index and lactylation level were decreased(P＜0.05).CONCLUSION:Lactoylation at the K197 site of PRDX1 promotes the proliferation and migration of glioblastoma cells.

Under the background of high-intensity fattening,probiotics have gradually become wide-ly used feed additives.It can inhibit the proliferation of pathogenic bacteria,maintain the microeco-logical balance in the digestive tract,and improve the host immune function.In order to screen lac-tic acid bacteria strains with potential probiotic effects,23 acid-producing strains were isolated from rumen fluid of healthy Duolang sheep by CaCO3-MRS solid medium.Safe strains were re-screened through primary screening and hemolysis tests.Acid tolerance tests and bile salt tests were used to screen the strains that were relatively adapted to the rumen environment.The survival rate of the strain M2 was 93.80％under pH3.0 and 59.72％under bile salt concentration of 0.30％and the survival rate is higher than other strains under high temperature conditions.Subsequently,morphological observation,molecular biological identification,testing of pathogen antagonism,antibiotic tolerance analysis,growth characteristics detection and other methods were used to fur-ther explore the characteristics of the strain.The results showed that the strain to be tested was Gram-positive bacillus.After 16S rRNA gene sequence comparison,the strain was identified as Lactobac illus salivus,which could be used as a candidate strain to develop probiotics.The research results laid a foundation for its application in Duolang sheep breeding.

This paper was aimed to study the effects of Huang-Lian Jie-Du decoction (HJD) on contents of oxidized low density lipoprotein (Ox-LDL),monocyte chemoattractant protein (MCP-1),vascular cell adhesion molecule (VCAM-1),malondialdehyde (MDA),and superoxide dismutase (SOD) of atherosclerosis (AS) rats' liver homogenate.Male SD rats were randomly divided into six groups,which were the normal control group,model group,positive control group (lovastatin),and the low-,middle-,high-dose HJD groups,with ten rats in each group.Except for the normal control group fed with normal diet,others were fed with high-fat diet,and regularly injected the vitamin D3.All groups were gavaged once daily from the 3rd week for 8 weeks until they were sacrificed.The results showed that compared with the normal control group,contents of MCP-1,VCAM-1 and Ox-LDL in the liver homogenate of the model group had significantly increased (p ＜ 0.05),after drug administration,all indexes mentioned above in the positive control group,the low-and middle-dose HJD group were decreased significantly (p ＜ 0.05);contents of MCP-1 and VCAM-1 of the high-dose HJD group decreased obviously (p ＜ 0.01);MDA content of liver homogenate increased significantly,after drug administration,contents of the positive control group and the low-dose HJD group decreased significantly (p ＜ 0.05);SOD content decreased significantly,after drug administration,contents of both the positive control group and the high-dose group increased significantly (p ＜ 0.05).It was concluded that HJD may play a role in AS intervention.Its mechanism may be related to its anti-inflammatory and antioxidant reaction.

BACKGROUND:Mechanical strain certainly has an effect on physiological activities of osteoblasts. Runx-2 is a target of bone morphogenic protein signal and is an important factor for regulation of osteoblastic differentiation. Bone morphogenic protein signal transduction pathway is involved in physiological response of osteoblast to stimulation of mechanical centrifugal force.
OBJECTIVE:To observe the effect of mechanical centrifugal force on bone morphogenetic protein signal pathway under different time period and speed.
METHODS:MC3T1-E1 cells were pre-treated in DMEM medium containing 10%fetal bovine serum for 24 hours, and then divided into control group, 90 r/min group, 180 r/min group and 250 r/min group. Each group was then subdivided into 6 hours, 12 hours and 24 hours centrifugation subgroups. Experiments were repeated for three times for different centrifugal speed and different time period. Except centrifugation, the control group was under the same environment. Total RNA was extracted and reversely transcribed into cDNA. Runx-2 gene expression was determined by real time fluorescent quantitative PCR.
RESULTS AND CONCLUSION:The expression of Runx-2 mRNA was increased with extension of time, showing a positive correlation between the two. The mRNA expression at 180 r/min was significantly higher than that at 90 r/min and 250 r/min (P<0.01);at 90 r/min and 180 r/min, the Runx-2 mRNA expression was higher than that in the control group (P=0.039), both of them showed significant difference along with the time. The difference of centrifugal force speed and duration is associated with different physiological response of osteoblasts in bone morphogenic protein signal pathway, which plays an important role in mechanical signal transmission and cascade reaction.

The prevalence of human and animal listeriosis for nearly 11 years in China was investigated in this study . The literature information about listeriosis in China from 2002 to 2012 was collected through retrieval system to make clinical and epidemiological statistical analysis of listeriosis .Cases of listeriosis were reported in 27 (79% ) provinces of China .The re-sult showed that animal listeriosis was reported for 123 times ,among these reports ,most were from pigs (39% ) ,and the sheep was in second place .Central nervous system infection was the main clinical manifestation of listeriosis in animals (72% ) . For human listeriosis ,84 clinical cases of listeriosis were reported ,including 35% cases in non-perinatal stage and 65% cases in perinatal stage .The main clinical manifestation of listeriosis was septicemia (51% ) .According to the result of investigation about listeriosis based on literatures information ,Listeriamonocytogenes caused humans and animals listeriosis annually ,which were reported in most provinces of China .The epidemic characteristics for listeriosis suggested that it was essential to strength-en the prevention and control of listeriosis .

A series of 29 cases of esophageal stenosis or obstruction caused by chemical burn or malignancy in the cervical or upper thoracic segment were treated by resection, and its continuity was re-established by a vascularized free or pedicled jejunal graft. It was successful in 28 cases, with only one failure. The age of these cases ranged from 4 to 65 years old. Among them, 4 children were between 4 and 6. Sixteen cases of pedicled jejunal grafts had their vessels anastomosed to cervical vessels at the upper end. The lengths of the grafts varied from 40 to 60 cm. In the 13 free jejunal grafts, jejunal blood vessels were anastomosed to the right gastroepiploic vessels and cervical vessels at the lower and upper ends, respectively, to re-establish circulation for long jejunal segments, while for the short jejunal grafts anastomosis was made to the cervical vessels only. The vascularized jejunal patch graft is an effective measure for repairing anastomotic fistula or localized wall defect. A regime of monitoring blood circulation of the jejunal transplant is described.