This study explored the effect of the Mycobacterium tuberculosis(Mtb)PE/PPE family protein PE_PGRS37 on the growth of Mycobacterium smegmatis(Ms)and macrophage autophagy during Mtb infection.The pe_pgrs37 gene was amplified from Mtb genome through PCR,and the recombinant vector pAIN-PE_PGRS37 was successfully constructed through homologous recombi-nation.pAIN-PE_PGRS37 and pAIN were integrated into Ms through electroshock to construct pAIN-PGRS37/Ms and pAIN/Ms re-combinant bacteria.Western blotting indicated that the PE_PGRS37 protein was correctly expressed in pAIN-PE_PGRS37/Ms.The re-combinant bacteria were inoculated in 7H9/7H10 medium,and their colony morphology and growth curves were observed.No signifi-cant difference in colony morphology was observed between pAIN-PE_PGRS37/Ms and pAIN/Ms.The growth rate significantly in-creased between 10 and 16 h,and a plateau was reached at 26 h.After infection of U937 cells with pAIN-PE_PGRS37/Ms and pAIN/Ms,macrophage autophagy flow was detected with western blotting and immunofluorescence.In the pAIN-PE_PGRS37/Ms-infected group,compared with the pAIN/Ms-infected group,macrophage LC3-II and p62 protein expression was significantly up-regulated(P<0.001)and inhibited autophagosome and lysosome fusion.The intracellular survival of the recombinant bacteria was detected through colony counting,and pAIN-PE_PGRS37/Ms showed significantly greater survival in macrophages at 12 h,24 h,and 48 h than pAIN/Ms(P<0.05).Our results suggested that PE_PGRS37 protein promotes Mycobacterium survival in macrophages by blocking macro-phage autophagy flow,thus inhibiting macrophage autophagy.

This study explored the effect of the Mycobacterium tuberculosis(Mtb)PE/PPE family protein PE_PGRS37 on the growth of Mycobacterium smegmatis(Ms)and macrophage autophagy during Mtb infection.The pe_pgrs37 gene was amplified from Mtb genome through PCR,and the recombinant vector pAIN-PE_PGRS37 was successfully constructed through homologous recombi-nation.pAIN-PE_PGRS37 and pAIN were integrated into Ms through electroshock to construct pAIN-PGRS37/Ms and pAIN/Ms re-combinant bacteria.Western blotting indicated that the PE_PGRS37 protein was correctly expressed in pAIN-PE_PGRS37/Ms.The re-combinant bacteria were inoculated in 7H9/7H10 medium,and their colony morphology and growth curves were observed.No signifi-cant difference in colony morphology was observed between pAIN-PE_PGRS37/Ms and pAIN/Ms.The growth rate significantly in-creased between 10 and 16 h,and a plateau was reached at 26 h.After infection of U937 cells with pAIN-PE_PGRS37/Ms and pAIN/Ms,macrophage autophagy flow was detected with western blotting and immunofluorescence.In the pAIN-PE_PGRS37/Ms-infected group,compared with the pAIN/Ms-infected group,macrophage LC3-II and p62 protein expression was significantly up-regulated(P<0.001)and inhibited autophagosome and lysosome fusion.The intracellular survival of the recombinant bacteria was detected through colony counting,and pAIN-PE_PGRS37/Ms showed significantly greater survival in macrophages at 12 h,24 h,and 48 h than pAIN/Ms(P<0.05).Our results suggested that PE_PGRS37 protein promotes Mycobacterium survival in macrophages by blocking macro-phage autophagy flow,thus inhibiting macrophage autophagy.

Objective To discuss the application of combined probiotics pretreatment and late triple therapy with helicobacter pylori infection in children.Methods 300 children with helicobacter pylori infection were randomly divided into three groups according to the way of random table.Group A was treated with standard triple therapy, omeprazole,amoxicillin and clarithromycin treatment,and 10 d was 1 treatment course.B group was given probiotic pretreatment,prior to the triple therapy using compound lactobacillus acidophilus piece 1 piece /time,cold water to take after meals,taking 2 weeks,follow -up treatment with triple therapy for 10d.In group C,triple therapy before using compound lactobacillus acidophilus piece 1 piece /time,cold water to take after meals,taking 2 weeks,follow -up treatment with triple therapy for 10d.To take one week after the completion of compound lactobacillus acidophilus piece 1 piece /time,cold water after meals.The clinical therapeutic effects were recorded.Results The helicobacter pylori clearance rate of group A was 55.00%,that of group B was 86.00%,that in group C was 89.00%,the helico-bacter pylori clearance rate of group B and group C was significantly higher than that in group A,the difference was statistically significant (χ2 =23.103 7,28.670 6,all P <0.05).The treatment effect between group B and group C had no statistically significant difference (χ2 =0.411 4,P >0.05 ).In A group,nausea and vomiting occurred in 5 cases,2 cases of diarrhea,abdominal distension abdominal pain in 4 cases,skin rash in 5 cases.In group B,nausea and vomiting occurred in 2 cases,0 cases of diarrhea,abdominal distension abdominal pain in 1 case,skin rash in 1 case.In group C,nausea and vomiting occurred in 1 case,0 cases of diarrhea,abdominal distension abdominal pain 0 cases,skin rashes in 1 case.The incidence rate of adverse reactions in group B and group C was lower than that in group A,the difference was statistically significant (χ2 =11.965 8,8.000 0,all P <0.05).The incidence rate of ad-verse reactions between group B and group C had no statistically significant difference (χ2 =0.687 3,P >0.05). Conclusion Joint probiotics pretreatment and late triple therapy application in helicobacter pylori in children can promote helicobacter pylori clearance,reduce the triple therapy drug adverse reactions,it is worthy of popularization and application in clinic.

Objective To investigate the predictive value of umbilical cord blood bilirubin for pathological jaundice in healthy term newborns. Methods Two ml navel string vein blood of baby were collected after giving birth in the normal newborn, and the hemobilirubin was detected by accidentally oxidation method. After birth, the infant's bilirubin level was tested on the forehead by the transcutaneous bilirubinometer at 8:00 -9:00 every morning until discharging from hospital. The ration of pathological jaundice of newborn and its treatment were analyzed in different levels of cord blood hemobilirubin. Results Fifty-nine cases ( 22.96% ) with pathological jaundice were diagnosed in 257 newboms.The concentration of cord blood hemobilirubin in baby with pathological jaundice [(39.68 ±8.10) μmol/L] was significantly higher than that of the normal newborn [(30.05 ±5.51) μmol/L](P＜0.01). As the concentration of cord blood hemobilirubin was increased, the incidence of pathological jaundice was raised (P＜ 0.01), and the cases that needed to intervention treatment was increased(P＜ 0.01). Conclusion The detection of the level of cord blood hemobilirubin is not only very worthy to estimate the occurrence of pathological jaundice of newborn, but also offer reliable evidence for clinical early diagnosis and treatment.