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Objective:To investigate the correlation between insulin resistance and alterations in gut microbiota using the animal model of insulin resistance(eLtaS transgenic mice).Methods:Glucose tolerance was measured in eLtaS trans mice and wild-type (WT) mice. Faecal samples of mice were collected for metagenomics and 16S rDNA sequencing. Alterations of gut microbiota in eLtaS trans mice were further analyzed. Faeces from eLtaS trans mice were transplanted into WT mice by " dirty cage" sharing experiment, and glucose tolerance of mice was measured at different time points after transplantation. Results:Significant differences in composition and function of gut microbiota were observed between eLtaS trans mice and WT mice( P=0.028). Compared with WT mice, the diversity of gut microbiota in eLtaS trans mice increased evidently, moreover the relative abundance of Phylum Firmicutes in eLtaS trans mice significantly increased( P<0.001). However, the relative abundance of Phylum Bacteroides and Phylum Verrucomicrobia decreased visibly( P=0.042, P=0.033). The relative abundance of Akkermansia muciniphila and Parabacterides distasonis related to metabolic diseases decreased significantly in eLtaS trans mice( P=0.033, P=0.013). The gut microbiota of eLtaS trans mice was clearly different from that of WT mice in carbohydrate metabolism, lipid metabolism, biosynthesis of other secondary metabolites, metabolism of other amino acids, energy production and transformation. The glucose tolerance of WT mice was impaired at 7th, 8th and 9th week after faecal transplantation, and recovered at 1 week after cessation of faecal transplantation. Conclusion:Insulin resistance leads to obvious changes of gut microbiota in mice, meanwhile the gut microbiota of insulin resistance mice can further induce impaired glucose tolerance.
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Objective:To prepare and identify a functional antibody FNA1 targeting the neuraminidase (NA) of influenza A virus N1 subtype.Methods:According to single-chain antibody fragment (scFv) sequence, the heavy chain and light chain variable region sequences of FNA1 were synthesized, and the recombinant expression plasmid pFRT-IgG1κ-FNA1 was constructed by linking the expression vector pFRT-IgG1κ. The FNA1 antibody was expressed in ExpiCHO cells and purified using affinity purification technique. The binding ability of FNA1 to the target proteins, influenza A virus N1 subtype NA antigens, was detected by ELISA. Flow cytometry was performed to analyze the binding ability of FNA1 to the NA antigens expressed on the surface of cell membrane. The in vitro activity of FNA1 against NA was evaluated by infecting 293T cells with pseudovirus. Results:Protein electrophoresis showed that FNA1 with high purity was obtained. FNA1 specifically recognized and bound to N1 subtype NA antigens in a concentration-dependent manner. FNA1 could effectively block NA activity by binding to N1 subtype NA protein expressed on the surface of cell membrane, thus inhibiting the release of packaged pseudovirus from cell surface and further inhibiting target cell infection.Conclusions:An antibody FNA1 targeting influenza A virus N1 subtype NA with in vitro functional activity was obtained.
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Objective:To prepare and identify a broad-spectrum antibody FHA3 targeting influenza A virus hemagglutinin (HA).Methods:According to the single-chain antibody fragment (scFv) sequence, the heavy chain (VH) and light chain (VL) variable regions of FHA3 were amplified by PCR and a recombinant plasmid pFRT-IgG1κ-FHA3 was constructed by linking the expression vector pFRT-IgG1κ. FHA3 was expressed in the ExpiCHO system and purified by affinity purification. The binding activity of FHA3 to influenza A virus HA was detected by ELISA. The neutralizing activity of FHA3 was detected in vitro by infecting host cells with pseudovirus. Results:SDS-PAGE showed that high-purity FHA3 was obtained. FHA3 could bind to H1N1 HA, H2N2 HA, H3N2 HA, H5N1 HA, H7N9 HA and H9N2 HA in a concentration-dependent manner. FHA3 had good neutralizing activity in vitro that was it could effectively block the invasion of H5N1 and H7N9 pseudoviruses into target cells at a low concentration of 5 μg/ml and H1N1 pseudovirus at 0.012 5 μg/ml. Conclusions:A broad spectrum antibody targeting HA protein of influenza A virus with neutralizing activity in vitro was obtained.
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Objective To explore the four tumor markers of alpha-fetoprotein ( AFP ), α-L-fucosidase( AFU), carbohydrate antigen 199 ( CA199) and carcinoembryonic antigen ( CEA) and their combined use for primary hepatocellular carcinoma (HCC) diagnosis and treatment value. Methods From February 2016 to August 2018,92 patients with primary hepatocellular carcinoma (HCC group),79 patients with benign liver disease (chronic hepatitis and cirrhosis group) and 99 healthy adults (control group) were selected as subjects. The serum levels of four tumor markers in different populations were compared. Results The serum levels of four tumor markers ( AFP ( 192. 4 ± 89. 3) μg／L、AFU( 78. 6 ± 25. 8) U／L、CA199 (107. 2 ± 59. 5) U／mL、 CEA ( 37. 9 ± 14. 9) μg／L) were significantly higher than those of benign liver disease group(AFP( 17. 4 ± 6. 3) μg／L、AFU( 35. 4 ± 17. 2) U／L、CA199( 29. 3± 15. 2) U／mL、CEA( 4. 9 ±1. 7) μg／L) and normal people( AFP(4. 8±1. 1) μg／L、AFU(12. 2±3. 6) U／L、CA199( 6. 4± 2. 3) U／mL、CEA(1. 8±0. 4) μg／L) . There differences had significant ( all P＜0. 05) . The abnormal rate of single factor in hepatocellular carcinoma group ( AFP 84. 8%, AFU 52. 2%, CA199 41. 3%, CEA35. 9%) was significantly higher than that in benign liver disease group ( AFP 15. 2%, AFU 19. 0%, CA19916. 5%, CEA13. 9%) and normal group (AFP 4. 0%,AFU 5. 0%,CA199 3. 0%,CEA 6. 0% ug／L),the difference was statistically significant ( all P＜0. 05) . The highest sensitivity was AFP ( 84. 8%) and the highest specificity was AFP and CA199 (91. 0%). The sensitivity of combined detection was 94. 6% higher than that of single index ( AFP 84. 8%, AFU52. 2%, CA199 41. 3%, CEA35. 9%) . Conclusion The combined detection of AFP,AFU,CA199 and CEA can increase the sensitivity of diagnosis of hepatocellular carcinoma and reduce the rate of missed diagnosis, which will be beneficial to the early diagnosis and treatment of hepatocellular carcinoma.
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Objective To investigate the clinical significance and changes of serum complement in lipid metabolism disorder in patients with fatty liver disease.Methods One hundred and forty patients with FLD from October 2015 to May 2017 were included in the study,in addition,120 patients with hyperlipidemia(the hyperlipidemia group)and 130 healthy subjects(the control group)in the same period were enrolled as controls.The differences in serum lipid,liver function enzymology,immunoglobulin,serum complement among the three groups were compared.Results There were significant differences in the levels of TC,TG,HDL-C,LDL-C,apoA and apoB among the three groups(TC:(5.7±1.6)mmol/L vs.(4.2±1.0)mmol/L vs.(3.5±1.1) mmol/L,F=105.01,P<0.05;TG:(2.8± 0.6)mmol/L vs.(1.5 ± 0.3)mmol/L vs.(1.1 ± 0.2)mmol/L,F=628.46,P<0.05;HDL-C:(1.2±0.3)mmol/L vs.(1.5±0.3)mmol/L vs.(1.8±0.4)mmol/L,F=107.10, P<0.05;LDL-C:(3.6±0.9)mmol/L vs.(3.0±0.8)mmol/L vs.(2.2±0.6)mmol/L,F=109.07,P<0.05;apoA:(1.0±0.2)g/L vs.(1.2±0.2)g/L vs.(1.4±0.3)g/L,F=95.20,P<0.05;apoB:(1.1±0.2)g/L vs.(0.9±0.2)g/L vs.(0.8±0.2)g/L,F=79.04,P<0.05).The levels of TC,TG,LDL-C and apoB in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of HDL-C and apoA in the FLD groups were significantly lower than those in the hyperlipidemia group and control group.The levels of TC,TG,LDL-C and apoB in the hyperlipidemia group were significantly higher than those in the control group.The levels of HDL-C and apoA in the hyperlipidemia group were significantly lower than those in the control group.There were significant differences in the levels of GGT,ALT,AST,IgG and IgM among the three groups(GGT:(77.4±15.3)U/L vs.(43.3±10.6)U/L vs.(25.5±8.2)U/L,F=668.12,P<0.05;ALT:(61.5±18.8)U/L vs.(35.7±11.2)U/L vs.(18.9±5.4)U/L,F=355.67,P<0.05;AST:(55.3±12.2)U/L vs.(32.4±12.5)U/L vs.(14.4±4.7)U/L,F=521.80,P<0.05;IgG:(15.7±3.9)g/L vs.(11.6±3.2)g/vs.,(8.5±2.6)g/L,F=162.34,P<0.05;IgM:(1.9±0.6)g/L vs.(1.2±0.4)g/L vs.(0.8±0.3)g/L,F=201.38,P<0.05).The levels of GGT,ALT,AST,IgG and IgM in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of GGT,ALT,AST,IgG and IgM in the hyperlipidemia group were significantly higher than those in the control group.There were significant differences in the levels of C3,C5,ASP and BF among the mild,moderate and severe fatty liver patients(C3:(2.1±0.4) g/L vs.(1.8±0.3)g/L vs.(1.0±0.2)g/L,F=436.37,P<0.05;C5:(92.3±10.7)mg/L vs.(71.8±8.8) mg/L vs.(58.9±6.5)mg/L,F=486.09,P<0.05; ASP:(51.4±6.8)nmol/L vs.(42.5±4.4)nmol/L vs.(32.8±5.2)nmol/L,F=369.29,P<0.05;BF:(0.48±0.13)g/L vs.(0.34±0.09)g/L vs.(0.23±0.04) g/L,F=233.39,P<0.05).The levels of C3,C5,ASP and BF in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of C3,C5,ASP and BF in the hyperlipidemia group were significantly higher than those in the control group.There were significant differences in the levels of C3,C5,ASP and BF among patients with mild,moderate and severe fatty liver disease(C3:(1.8±0.3)g/L vs.(2.1±0.4)g/L vs.(2.5±0.4)g/L,F=30.85,P<0.05;C5:(80.5±9.6)mg/L vs.(92.3±10.5)mg/L vs.(100.7±8.)mg/L,F=39.39,P<0.05; ASP:(42.4±6.3)nmol/L vs.(52.8±5.7)nmol/L vs.(61.9±5.6) nmol/L,F=98.19,P<0.05;BF:(0.33±0.12)g/L vs.(0.45±0.11)g/L vs.(0.57±0.09)g/L,F=41.26,P<0.05).The levels of C3,C5,ASP and BF in the mild FLD patients were significantly lower than those in moderate and severe FLD patients.The levels of C3,C5,ASP and BF in the moderate FLD patients were significantly lower than those in severe FLD patients.Conclusion The detection of C3,C5,ASP and BF levels based on routine testes has important clinical value for the assessment of the condition,the treatment and the prognosis of FLD patients.
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Objective To investigate the clinical value of joint detection of six tumor markers in patients with colorectal cancer. Methods Eighty?six patients with colorectal cancer were included in the study group,86 healthy subjects were selected as the control group at the same period. The difference of tumor markers in different groups,tumor stages and prognosis were compared. Results The levels of carcinoembryonic antigen (CEA),carbohydrate antigen 19?9 (CA19?9),carbohydrate antigen 242 (CA242),carbohydrate antigen 72?4 ( CA72?4) , carbohydrate antigen 125 ( CA125 ) and carbohydrate antigen 50 ( CA50 ) in study group were significantly higher than those in the control group (CEA: (22. 5±6. 2)μg/L vs. (2. 2±1. 0)μg/L;CA19?9:(95. 7±27. 3) U/ml vs. (17. 1±9. 5) U/ml;CA242:(29. 5±8. 3) U/ml vs. (6. 0±2. 7) U/ml;CA72?4:(21. 6 ±5. 1) U/ml vs. (3. 6±1. 2) U/ml;CA125:(95. 4±32. 8) U/ml vs. (18. 9±8. 4) U/ml;CA50:(51. 8±20. 6)μg/L vs. (8. 3±3. 7)μg/L,t=29. 98,25. 22,24. 97,31. 86,20. 95,19. 27,P<0. 05). Among the single index detections,the sensitivity and negative predictive value of CA72?4 were the highest ( 61. 6%, 68. 3%) , the specificity of CA19?9 was the highest( 91. 9%) ,the positive predictive value of CEA was the highest ( 80. 4%) . The sensitivity,positive predictive value and negative predictive value of the joint detection were all higher than those in each single index detection (80. 3%,87. 3%,74. 1%). The levels of CEA,CA19?9,CA242,CA72?4, CA125 and CA50 in patients with stage III and IV were significantly higher than those in patients with stageⅠandⅡ(CEA:(32. 7±7. 1)μg/L vs. (15. 9±4. 4)μg/L;CA19?9:(127. 8±33. 7) U/ml vs. (52. 5±13. 8) U/ml;CA242:(40. 3±12. 7) U/ml vs. (23. 5±8. 6) U/ml;CA72?4:(37. 6±10. 2) U/ml vs. (13. 6±4. 1) U/ml;CA125:(128. 9±38. 4) U/ml vs. (59. 7±12. 8) U/ml;CA50:(88. 3±23. 7)μg/L vs. (41. 8±15. 6)μg/L,t=13. 04,13. 32,7. 11,14. 06,10. 99,10. 64,P<0. 05) . The levels of CEA,CA19?9,CA242,CA72?4,CA125 and CA50 in the recurrent metastasis group were significantly higher than those in the non?recurrent metastasis group ( CEA:( 37. 7 ± 8. 6 ) μg/L vs. ( 3. 8 ± 1. 7 ) μg/L;CA19?9:( 110. 5 ± 29. 4 ) U/ml vs. ( 25. 5 ± 13. 8 ) U/ml;CA242:( 33. 6 ± 10. 3 ) U/ml vs. ( 15. 5 ± 6. 6 ) U/ml;CA72?4:( 33. 1 ± 15. 3 ) U/ml vs. ( 9. 3 ± 3. 0 ) U/ml;CA125:(113. 4±31. 7) U/ml vs. (28. 7±7. 8) U/ml;CA50:(55. 4±14. 6)μg/L vs. (16. 8±9. 6)μg/L,t=29. 04,18. 31,9. 86,11. 47,19. 28,14. 65,P<0. 05) . Conclusion The joint detection of six markers can further improve the sensitivity, positive predictive value and negative predictive value of diagnosis, and can provide a more reliable basis for the auxiliary diagnosis of colorectal cancer.
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Objective To investigate the clinical significance and changes of serum complement in lipid metabolism disorder in patients with fatty liver disease.Methods One hundred and forty patients with FLD from October 2015 to May 2017 were included in the study,in addition,120 patients with hyperlipidemia(the hyperlipidemia group)and 130 healthy subjects(the control group)in the same period were enrolled as controls.The differences in serum lipid,liver function enzymology,immunoglobulin,serum complement among the three groups were compared.Results There were significant differences in the levels of TC,TG,HDL-C,LDL-C,apoA and apoB among the three groups(TC:(5.7±1.6)mmol/L vs.(4.2±1.0)mmol/L vs.(3.5±1.1) mmol/L,F=105.01,P<0.05;TG:(2.8± 0.6)mmol/L vs.(1.5 ± 0.3)mmol/L vs.(1.1 ± 0.2)mmol/L,F=628.46,P<0.05;HDL-C:(1.2±0.3)mmol/L vs.(1.5±0.3)mmol/L vs.(1.8±0.4)mmol/L,F=107.10, P<0.05;LDL-C:(3.6±0.9)mmol/L vs.(3.0±0.8)mmol/L vs.(2.2±0.6)mmol/L,F=109.07,P<0.05;apoA:(1.0±0.2)g/L vs.(1.2±0.2)g/L vs.(1.4±0.3)g/L,F=95.20,P<0.05;apoB:(1.1±0.2)g/L vs.(0.9±0.2)g/L vs.(0.8±0.2)g/L,F=79.04,P<0.05).The levels of TC,TG,LDL-C and apoB in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of HDL-C and apoA in the FLD groups were significantly lower than those in the hyperlipidemia group and control group.The levels of TC,TG,LDL-C and apoB in the hyperlipidemia group were significantly higher than those in the control group.The levels of HDL-C and apoA in the hyperlipidemia group were significantly lower than those in the control group.There were significant differences in the levels of GGT,ALT,AST,IgG and IgM among the three groups(GGT:(77.4±15.3)U/L vs.(43.3±10.6)U/L vs.(25.5±8.2)U/L,F=668.12,P<0.05;ALT:(61.5±18.8)U/L vs.(35.7±11.2)U/L vs.(18.9±5.4)U/L,F=355.67,P<0.05;AST:(55.3±12.2)U/L vs.(32.4±12.5)U/L vs.(14.4±4.7)U/L,F=521.80,P<0.05;IgG:(15.7±3.9)g/L vs.(11.6±3.2)g/vs.,(8.5±2.6)g/L,F=162.34,P<0.05;IgM:(1.9±0.6)g/L vs.(1.2±0.4)g/L vs.(0.8±0.3)g/L,F=201.38,P<0.05).The levels of GGT,ALT,AST,IgG and IgM in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of GGT,ALT,AST,IgG and IgM in the hyperlipidemia group were significantly higher than those in the control group.There were significant differences in the levels of C3,C5,ASP and BF among the mild,moderate and severe fatty liver patients(C3:(2.1±0.4) g/L vs.(1.8±0.3)g/L vs.(1.0±0.2)g/L,F=436.37,P<0.05;C5:(92.3±10.7)mg/L vs.(71.8±8.8) mg/L vs.(58.9±6.5)mg/L,F=486.09,P<0.05; ASP:(51.4±6.8)nmol/L vs.(42.5±4.4)nmol/L vs.(32.8±5.2)nmol/L,F=369.29,P<0.05;BF:(0.48±0.13)g/L vs.(0.34±0.09)g/L vs.(0.23±0.04) g/L,F=233.39,P<0.05).The levels of C3,C5,ASP and BF in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of C3,C5,ASP and BF in the hyperlipidemia group were significantly higher than those in the control group.There were significant differences in the levels of C3,C5,ASP and BF among patients with mild,moderate and severe fatty liver disease(C3:(1.8±0.3)g/L vs.(2.1±0.4)g/L vs.(2.5±0.4)g/L,F=30.85,P<0.05;C5:(80.5±9.6)mg/L vs.(92.3±10.5)mg/L vs.(100.7±8.)mg/L,F=39.39,P<0.05; ASP:(42.4±6.3)nmol/L vs.(52.8±5.7)nmol/L vs.(61.9±5.6) nmol/L,F=98.19,P<0.05;BF:(0.33±0.12)g/L vs.(0.45±0.11)g/L vs.(0.57±0.09)g/L,F=41.26,P<0.05).The levels of C3,C5,ASP and BF in the mild FLD patients were significantly lower than those in moderate and severe FLD patients.The levels of C3,C5,ASP and BF in the moderate FLD patients were significantly lower than those in severe FLD patients.Conclusion The detection of C3,C5,ASP and BF levels based on routine testes has important clinical value for the assessment of the condition,the treatment and the prognosis of FLD patients.
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Objective To clone and express the alkaline protease AprA , one important virulence factor secreted by Pseudomonas aeruginosa(PAE)in Escherichia coli, to clone and express the inhibitor of AprA (AprI) and its substrate flagellin , and to detect the function of AprA and the inhibitory function of AprI .Methods The genes encoding AprA ,AprI and flagellin gene were amplified respectively by PCR using PAE PAO 1 genome DNA as the template .The expression vec-tors (pET-28a-AprA, pET-28a-AprI and pET-28a-Flagellin) were constructed and transformed into E.coli BL21(DE3) respectively.The recombinant AprA protein was expressed by IPTG induction and purified via denaturing and renaturation. The recombinant AprI and flagellin were expressed and purified by Ni 2+affinity chromatography .The cleavage activities of AprA on flagellin were detected by SDS-PAGE.Results Recombinant AprA , AprI and flagellin protein were expressed and purified .It was demonstrated that AprA cleaved flagellin , which was blocked by AprI .Conclusion Recombinant AprA could cleave its substrates as an alkaline protease , and its inhibitor AprI inhibits the activities of AprA .This study will contribute to further investigations on the role of AprA in the pathogenesis of PAE .
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Objective To express the beta hemolysin ( Hlb), an important toxin secreted by Staphylococcus aureus ( S.aureus) and the mutant protein Hlb H-149-N , to detect the hemolytic activity of Hlb and Hlb H-149-N on sheep erythrocytes , and to prepare the specific antibodies against Hlb which can inhibit the hemolytic activity of Hlb .Methods Hlb gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as template.The expression vector pET-28a-hlb was constructed and transformed into E.coli BL21(DE3).The expression vector pET28a-hlbH-149-Nwas constructed through point mutation.The recombinant Hlb and Hlb H-149-N protein were expressed and purified by Ni 2+affinity chromatography .The hemolytic activity of Hlb and Hlb H-149-N was measured by sheep erythrocyte lysis assay .Results Recombinant Hlb protein and the mutant were obtained .Further investigations showed that Hlb could significantly induce the lysis of SRBC while HlbH-149-N could not.The specific polyclonal antibodies against Hlb (anti-Hlb) were prepared.It was found that anti-Hlb recognized Hlb and Hlb H-149-N .Moreover , it was found that anti-Hlb blocked the hemolytic activity of Hlb .Conclusion The recombinant Hlb protein with high hemolytic activity and Hlb H-149-N without hemolytic activity are obtained while its neutralized antibody is pepared .Hlb from S.aureus has different hemolytic effects on erythrocytes from various species .Our findings will facilitate the investigation on the role of Hlb in the pathogenesis of S.aureus.
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OBJECTIVE To discuss the risk factors and countermeasures of hospital infection in department of laboratory. METHODS The risk factors existing in department of laboratory for hospital infection were imestigated and corresponding measures and strategies were set up. RESULTS By adopting corresponding measures and strategies, hospital infection in department of laboratory was controlled and prevented. CONCLUSIONS The occurrence of hospital infection in the department of laboratory can be effectively prevented, through the establishment of complete system, strict operation of the rules and corresponding control measures.
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0.05). The incidence of restenosis and major adverse cardiovascular events were significantly less in the hyperbaric oxygen group than that in the control group (6.67% vs 22.58% for restenosis, and 8.82% vs 38.24% for major adverse cardiovascular events; P0.05). No severe adverse effect was found during hyperbaric oxygen therapy in the hyperbaric oxygen group. Conclusion Hyperbaric oxygen therapy is effective and safe in preventing restenosis after intracoronary stenting.
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Objective To discuss the emergency interventional therapy for ureter invasion by CACX, obstructive bilateral hydronephrosis and uremia complicated with hemorrhoea. Methods 5 cases with ureter invasion by CACX, obstructive bilateral hydronephrosis and uremia suffered from hemorrhoea during hemodialysis, who were performed with emergency interventional therapy. Results After therapy, hemostasis were realized in all cases, and all symptoms were alleviated, such as vaginal fluid and fall-swell in pelvis. The short-term total effective rate was 100% . Conclusion Interventional chemoembolization can be used in the treatment of CACX with acute hemorrhoea.
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Objective To analyze the reasons of CT misdiagnosis of solid-pseudopapillary tumor of pancreas (SPTP), including wrong orientation, mislocation. Methods The literatures on SPTP were discussed. Results Lesions of SPTP could appear at any position of the pancreas. Conclusion The result is very significant for guiding treatment and judging prognosis.
