Effect of different abutment materials on the expression of genes and proteins related to hemidesmosome adhesion in human gingival epithelial cells / 中华口腔医学杂志

Objective: To investigate the effects of polyetheretherketone, zirconium dioxide, and titanium abutment materials on the expression of genes and proteins related to hemidesmosome adhesion in human gingival epithelial cells, in order to screen out abutment materials that are easier for epithelial adhesion. Methods: Forty-eight specimens were prepared in each of the three materials, polyetheretherketone, zirconium oxide, and pure titanium specimens. The surface morphology of each group of specimens was observed by scanning electron microscopy, the surface roughness was measured by the white light interferometer, and the contact angle was measured by optical contact angle measuring instrument. The early adhesion status of human gingival epithelial cells on the surface of each group of specimens was observed by scanning electron microscopy, and the proliferation ability of human gingival epithelial cells on the surface of each group of specimens was assessed by using a cell counting kit, and the expression levels of genes and proteins related to the adhesion of human gingival epithelial cells on the surface of each group of specimens were detected by real-time fluorescence quantitative PCR and Western blotting, respectively. Results: The surface morphology of the three groups of specimens was flat and smooth. The mean roughness (Ra value) of the polyetheretherketone, zirconia, and pure titanium groups were (95.63±2.06), (37.93±3.56), and (134.2±4.62) nm (F=368.16, P<0.001), respectively, and the mean maximum height (Rz value) was (2.42±0.22), (0.87±0.10) and (3.77±0.28) nm (F=91.95, P<0.001), with statistical significance (P<0.05). The contact angles were 81.23°±0.91°, 82.08°±2.10°, and 80.47°±1.85°, respectively, with no statistically significant overall difference (F=0.45, P=0.658). Human gingival epithelial cells showed irregular shapes such as flattened and extended polygons and polygons on the surface of the three groups of specimens, exhibiting a typical paving stone pattern. The differences in cell proliferation among the three groups at 1 and 3 d of culture were not statistically significant (P>0.05). Cell proliferation in the polyetheretherketone group was significantly greater those those in the zirconia and pure titanium groups at 5 and 7 d of culture (P<0.05). The mRNA expression levels and protein expression levels of laminin α3, integrin β4, and collagen ⅩⅦ in the polyetheretheretherketone group at 3 and 7 d of incubation were significantly greater than those in the zirconium oxide and pure titanium groups at the same time points (P<0.05). Conclusions: Polyetheretherketone is more conducive to the adhesion of hemidesmosome in human gingival epithelial cells than zirconium dioxide and pure titanium abutment materials.

Effect of type 2 diabetes mellitus on mandibular bone regeneration and the expression of T helper cell 17/regulat-ory T cell-related factors in mice / 华西口腔医学杂志

OBJECTIVES@#To observe the effect of type 2 diabetes mellitus (T2DM) on mandibular bone regeneration and the expression of factors related to T helper cell 17 (Th17 cell) and regulatory T cell (Treg cell) in mice.@*METHODS@#Thirty-six 6-week-old C57BL/6J male mice were randomly divided into normal control (NC) and T2DM groups. Fasting blood glucose levels were detected 0 d, 7 d, 14 d, and 28 d after surgery for mandibular defects. Hematoxylin-eosin (HE) staining was used in observing the bone after 7 d, 14 d, and 28 d of the healing process. Immunohistochemical staining was used in observing the expression of alkaline phosphatase (ALP), Runt-related transcription factor 2 (RUNX2), forkhead box protein P3 (Foxp3), retinoic acid related orphan receptor gamma T (RORγt), and protein tyrosine phosphatase non-receptor type 2 (PTPN2) after 7 d, 14 d, and 28 d of healing.@*RESULTS@#HE staining showed that the area with new bones in the T2DM group was significantly smaller than that in the NC group. Immunohistochemical staining showed that the expression of osteogenesis related proteins ALP and RUNX2 were significantly reduced in the T2DM group. In addition, the number of RORγt positive cells increased, whereas the number of Foxp3 positive cells and the expression PTPN2 decreased significantly in the mandibular bone defect in mice with T2DM.@*CONCLUSIONS@#T2DM significantly inhibit mandibular bone regeneration in mice. Decline in PTPN2 expression and the transition of Treg and Th17 may be the underlying molecular mechanisms.

Rapid detection of Chinese herbal medicine dyed by auramine O, amaranth and sunset yellow by ion mobility spectrometry / 中国药学杂志

OBJECTIVE: To develop a fast detecting method for three dyes (auramine O, amaranth and sunset yellow) illegally added into Chinese herbal medicine by ion mobility spectrometry (IMS). METHODS: The analysis was performed on ion mobility spectrometry, with source voltage 1 800 V for negative source and 2 300 V for positive source, drift tube voltage 7 500 V, gas inlet temperature 180℃, drift tube temperature 180℃, gate voltage 45 V, gate pulse width 120 μs, drift gas flow 1.2 L·min-1, exhaust pump 0.8 L·min-1, run time 30 s and spectrum length 25 ms. The samples were extracted by methanol, and then injected into the IMS system. The judgement of whether dye was added or not was made by comparing the migration time of the test samples with that of the reference substances. RESULTS: The auramine O, amaranth and sunset yellow could be rapidly identified. The minimum detection concentration of each compound was determined according to the signal to noise ratio (S/N) of 3. CONCLUSION: The IMS method for detecting auramine O, amaranth and sunset yellow illegally added into Chinese herbal medicine is simple, rapid and expected to be used as an initial screening method in drug rapid detecting system.

A study on the influence of montelukast on chromogranin A protein and mRNA expressions at lung tissue of asthmatic rat / 预防医学

Objective To investigate the potential roles of chromogranin A in pathogenesis of asthmatic inflammation,and to assess the regulation of montelukast on chromogranin A expression.Methods The rat asthma model was established with ovalbumin,and they were allocated to three groups,named asthma group,control group and montelukast group.The expressions of chromogranin A protein and mRNA at lung tissue were detected by immunohistochemisty or real-time PCR methods,and positive expression intensity of chromogranin A protein was assayed by optical density.The correlation between chromogranin A protein and mRNA was also analyzed.Results The expression levels of chromogranin A protein in asthma group(0.34 ±0.05 optical density)was significantly higher than that in control group (0.21 ±0.06 optical density)(P<0.01 ).The expression levels of chromogranin A mRNA in asthma group (4.02 ±0.95 relative quantity value)was significantly higher than that in control group(P<0.01 ).The expression levels of chromogranin A protein in montelukast group(0.28 ±0.04 optical density)was dramatically lower than that in asthma group (0.34 ±0.05 optical density)(P<0.05),while there were no statistical significance of chromogranin A mRNA(3.67 ±0.78 relative quantity value)between those two groups(P>0.05 ).But levels of mRNA was positively correlated with protein of chromogranin A (r=0.635,P<0.01).Conclusion Expressions of chromogranin A protein and mRNA at lung tissue were increased in asthmatic rats,and the results demonstrated that chromogranin A perhaps participated in the pathogenesis of asthma inflammation,but this function of chromogranin A protein could be down regulated by montelukast.

Expression of Serum HMGB-1 in Patients with Secondary Hemo-phagocytic Lymphohistiocytosis and Its Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression levels and clinical significance of serum high mobility group box 1 (HMGB-1) in patients with secondary hemophagocytic lymphohistiocytosis (sHLH).</p><p><b>METHODS</b>Serum HMGB-1 levels were determined by using enzyme linked immunosorbent assays (ELISA) in 51 sHLH patients and 15 healthy contrlols. Other laboratory data, including soluble interleukin-2 receptor (sCD25), white blood cells (WBC), hemoglobin (Hb), platelet (Plt), fibrinogen (FIB), lactate dehydrogenase (LDH), triglyceride (TG), alanine transaminase (ALT), aspartate aminotransferase (AST), serum ferritin (SF), C reactive protein (CRP), and blood sedimentation rate (ESR) were also collected.</p><p><b>RESULTS</b>Serum HMGB-1 levels in the newly diagnosed group were significantly higher than that in the control group (P<0.01). Serum HMGB-1 levels in the newly diagnosed lymphoma-associated HLH (LHLH) group were significantly higher than that in non-HLH group, including infection-associated HLH (IHLH) and autoimmune-associated HLH (AHLH) group (P<0.05); The serum HMGB-1 levels in the clinical remission group were significantly lower than that in the newly diagnosed group (P<0.05), however, serum HMGB-1 was not decreased significantly in the progression/relapsed group, compared with the newly diagnosed group (P>0.05). Serum HMGB-1 levels in newly diagnosed sHLH patients positively correlated with sCD25 (r=0.62, P<0.01) and ESR (r=0.55, P<0.05). The receiver operating characteristic curves (ROC) for serum HMGB-1 levels of sHLH patients and healthy controls produced a cutoff value at 15.3 µg/L, with its 90% sensitivity and 99% specificity, respectively. In addition, an optimal cutoff value for HMGB-1 was 27.4 µg/L in the patients LHLH and non-HLH (AHLH+IHLH) with 96% sensitivity and 81% specificity, separately.</p><p><b>CONCLUSION</b>Serum HMGB-1 levels possesses an important clinically significance for disease diagnosis, differential diagnosis, evaluation of nosographic activity and treatment efficacy in the patients with sHLH.</p>

Oxidative damage of NIH/3T3 cells induced by nickel smelting fumes / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the cytotoxicity and oxidative damage on NIH/3T3 cells induced by nickel smelting fume.</p><p><b>METHODS</b>NIH/3T3 cells were treated with nickel smelting fume collected from a nickel smelting factory in China with doses of 0, 6.25, 12.50, 25.00, 50.00, and 100.00 µg/ml for 6 h. Dose-dependent cytotoxicity in cells were assessed by Cell Counting Kit-8 (CCK-8), natural red uptake assay, and lactate dehydrogenase (LDH) leakage assay, and the level of oxidative damage was assessed based on the activity of catalase (CAT), percentage inhibition of superoxide dismutase (SOD), and content of malonaldehyde (MDA).</p><p><b>RESULTS</b>The relative survival of NIH/3T3 cells decreased with the increase in the dose of nickel smelting fume. In the CCK-8 assay, the group with 100 µg/ml nickel smelting fume showed a cell growth inhibition rate of 86%, with a significant difference compared with the control group (P < 0.05). LDH activity increased with increasing dose of nickel smelting fume: the groups of 12.50, 25, 50, and 100 µg/ml nickel smelting fume all showed increased LDH activities as compared with the control group (P < 0.05). The activities of CAT were significantly reduced in groups of 25, 50, and 100 µg/ml nickel smelting fume as compared with that of the control group (P < 0.05). As the dose of nickel smelting fume increased, the percentage inhibition of SOD and the content of MDA increased, with significant differences compared with the control group (P < 0.05).</p><p><b>CONCLUSION</b>Oxidative damage may be induced in NIH/3T3 cells after 6 h of exposure to nickel smelting fume, which leads to cell death.</p>

Electroacupuncture at Zusanli (ST 36) for treatment of nausea and vomiting caused by the chemotherapy of the malignant tumor: a multicentral randomized controlled trial / 中国针灸

<p><b>OBJECTIVE</b>To compare the clinical effects between electroacupuncture at Zusanli (ST 36) combined with intravenous drip of Granisetron and intravenous drip of Granisetron only for treatment of nausea and vomiting caused by the chemotherapy of the malignant tumor.</p><p><b>METHODS</b>The methods of multicentral, randomized controlled trial were used, the observation group (127 cases) was treated with electroacupuncture at Zusanli (ST 36) combined with intravenous drip of Granisetron, and the control group (119 cases) was treated with intravenous drip of Granisetron only.</p><p><b>RESULTS</b>The total effective rate of 90.5% in observation group was superior to that of 84.0% in control group (P < 0.01); the nausea and vomiting scores of two groups were obviously decreased after treatment (both P < 0.001), and the decreased degree of the observation group was superior to that of control group (P < 0.001).</p><p><b>CONCLUSION</b>Electroacupuncture at Zusanli (ST 36) can significantly alleviate the symptoms such as nausea and vomiting caused by the chemotherapy of the patients.</p>

Prokaryotic expression of trigeminy artificial fusion gene of Leptospira interrogans and the immunogenicity of its products / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct lipL32/1-lipL21-OmpL1/2 fusion gene of Leptospira interrogans and its prokaryotic expression system, and to identify the immunogenicity of its products.</p><p><b>METHODS</b>PCR using linking primers was applied to construct lipL32/1-lipL21-OmpL1/2 fusion gene and a prokaryotic expression system of the fusion gene was then established using routine genetic engineering technique. SDS-PAGE was used to examine output of the target recombinant protein rLipL32/1-LipL21-OmpL1/2. Double immunodiffusion and Western Blot assay were applied to identify immunogenicity of rLipL32/1-LipL21-OmpL1/2.</p><p><b>RESULT</b>lipL32/1-lipL21-OmpL1/2 fusion gene with correct sequence and its prokaryotic expression system E.coli BL21DE3pET42a-lipL32/1-lipL21-ompL1/2 was obtained in this study. The output of rLipL32/1-LipL21- OmpL1/2 after optimisation was 37.78 mg/L. The immunodiffusion titer of rabbit antiserum against rLipL32/1-LipL21-OmpL1/2 was 1:4. The rLipL32/1-LipL21-OmpL1/2 antiserum was able to recognize rLipL32/1-LipL21-OmpL1/2, rLipL32/1, rLipL21 and rOmpL1/2. Positive Western hybridization signals were found among rLipL32/1-LipL21-OmpL1/2 and rabbit antiserum against whole cell of strain 56601 and serum from patients infected with L.interrogans serogroups Icterohaemorrhagiae, Grippotyphosa, Autumnalis and Pomona.</p><p><b>CONCLUSION</b>The fusion gene lipL32/1-lipL21-OmpL1/2 and its prokaryotic expression system were successfully constructed in this study. The expressed fusion protein can be used as the antigen for developing universal genetic engineering vaccine and universal serological tests of leptospirosis.</p>

Reconstruction of Leptospira interrogans lipL21 gene and characteristics of its expression product / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To reconstruct the nucleotide sequence of Leptospira interrogans lipL21 gene for increasing the output of prokaryotic expression and to understand the changes on immunogenicity of the expression products before and after reconstruction, and to determine the position of envelope lipoprotein LipL21 on the surface of leptospiral body.</p><p><b>METHODS</b>According to the preferred codons of E.coli, the nucleotide sequence of lipL21 gene was designed and synthesized, and then its prokaryotic expression system was constructed. By using SDS-PAGE plus BioRad agarose image analysor, the expression level changes of lipL21 genes before and after reconstruction were measured. A Western blot assay using rabbit anti-TR/Patoc I serum as the first antibody was performed to identify the immunoreactivity of the two target recombinant proteins rLipL21s before and after reconstruction. The changes of cross agglutination titers of antisera against two rLipL21s before and after reconstruction to the different leptospiral serogroups were demonstrated using microscope agglutination test (MAT). Immuno-electronmicroscopy was applied to confirm the location of LipL21s.</p><p><b>RESULT</b>The expression outputs of original and reconstructed lipL21 genes were 8.5 % and 46.5 % of the total bacterial proteins, respectively. Both the two rLipL21s could take place immune conjugation reaction with TR/Patoc I antiserum. After immunization with each of the two rLipL21s in rabbits, the animals could produce specific antibody. Similar MAT titers with 1:80 - 1:320 of the two antisera against rLipL21s were present. LipL21 was confirmed to locate on the surface of leptospiral envelope.</p><p><b>CONCLUSION</b>LipL21 is a superficial antigen of Leptospira interrogans. The expression output of the reconstructed lipL21 gene is remarkably increased. The expression rLipL21 maintains fine antigenicity and immunoreactivity and its antibody still shows an extensive cross immunoagglutination activity. The high expression of the reconstructed lipL21 gene will offer a favorable condition to use its product for further developing a novel universal vaccine as well as detection kit of leptospirosis.</p>

Study on the immunogenicity of major leptospiral genus-specific protein antigens and the distribution of antigens in different serogroups of Leptospira interrogans / 中华流行病学杂志

<p><b>OBJECTIVE</b>The determination of antigenicity and immunogenicity of Leptospira interrogans genus-specific outer envelope proteins (OEPs) will offer evidence for developing universal leptospiral genetic engineering vaccine and detection kit.</p><p><b>METHODS</b>In this study, Ni-NTA affinity chromatography is used to purify the recombinant products rLipL21, rOmpL1/1, rOmpL1/2, rLipL32/1, rLipL32/2, rLipL41/1 and rLipL41/2 expressed by the major genotypes of four leptospiral OEPs of 15 serogroups. SDS-PAGE is applied to examine the expression and purity of the recombinant proteins. Rabbits are intracutaneously immunized with the recombinant proteins to obtain antisera. Microscope agglutination test (MAT) is used to measure the cross inmmunoagglutination titers of antisera. The OMPs of the reference standard strains belonging to 15 serogroups of L. interrogans in China and L. biflexa strain Patoc I are prepared using salt-denature method. By each of the antisera as the first antibody, Western blot assay is established to detect the natural expressions and immunoreactivity of the four OEPs.</p><p><b>RESULTS</b>The outputs of rLipL21, rLipL32/1, rLipL32/2, rLipL41/1l, rLipL41/2, rOmpL1/1 and rOmpL1/2 are 10%, 40%, 35%, 15%, 10%, 30% and 15%, respectively. Each the purified recombinant proteins shows a single fragment after SDS-PAGE. Each the rabbit antisera displays extensive cross immunoreactivity between the products expressed by different genotypes of the same gene and the MAT titers ranging from 1:2-1:128. All the four OEPs can be detectable in the OEPs preparations. However, LipL21 is found to exist only in L. interrrogans.</p><p><b>CONCLUSION</b>The results of this study indicate that all the four OEPs are superficial genus-specific antigens of Leptospira which can be used as the candidate antigens of leptospiral universal vaccine and detection kit.</p>

The cardioprotective effect and mechanism of lumbrokinase / 药学学报

<p><b>AIM</b>To investigate the protective effect of lumbrokinase against myocardial ischemia and to further explore its underlying mechanisms.</p><p><b>METHODS</b>The effect of lumbrokinase on myocardial ischemia was observed by a model of acute myocardial infarction due to permanent ligation of the left anterior descending coronary artery in rats. Patch-clamp technique and laser scanning confocal microscopy were utilized to study the action of lumbrokinase on L-type calcium current (ICa-L) and intracellular calcium concentration ([Ca2+]i).</p><p><b>RESULTS</b>Lumbrokinase decreased the infarct size of myocardium in a dose-dependent manner. The inhibitory rate of lumbrokinase at the dose of 20, 40 and 80 mg x kg(-1) was 7.7%, 34.6% and 46.2%, respectively. The electrophysiological studies displayed that, at + 10 mV, the ICa-L was markedly reduced from (-14.42 +/- 1.53) pA/pF to (-11.33 +/- 1.40) pA/pF (decreased by 21.4%, P <0.01) and (-9.92 +/- 1.31) pA/pF (decreased by 36.5%, P <0.01) by lumbrokinase (10 and 50 micromol x L(-1)), respectively. Confocal experiments showed that 10 micromol x L(-1) lumbrokinase showed no obvious effects on [Ca2+]i at resting states (P > 0.05). However, the increase of [Ca2+]i induced by 60 mmol x L(-1) KCl was distinctly limited by 10 micromol x L(-1) lumbrokinase (P <0.01). Within 240 s, the no obvious peak value of fluorescent intensity (FI) was shown.</p><p><b>CONCLUSION</b>Lumbrokinase showed protective action against myocardial infarction in rats. The possible mechanisms of anti-ischemia could be attributed to decreasing ICa-L and [Ca2+] of ventricular myocytes in rats.</p>

M3-R/IK(M3)--a new target of antiarrhythmic agents / 药学学报

<p><b>AIM</b>To investigate the relationship between M3-R/IK(M3) and arrhythmia in order to find a new target for antiarrhythmic agents.</p><p><b>METHODS</b>Using the acute ischemic model of rats and patch-clamp techniques, the effects of the M3 receptor on the occurrence of arrhythmias and its possible mechanisms were studied.</p><p><b>RESULTS</b>In acute ischemic model of rats, the M3 receptor antagonist 4-diphenylacetoxy-N-methylpiperidine-methiodide (4DAMP) increased the occurrence of arrhythmias, and the M3 receptor agonist choline suppressed the onset and the development of arrhythmias (P < 0. 01). No change was observed after treatment with other receptor antagonists (M1, M2, and M4). With patch-clamp techniques, it was found that choline induced K+ current could be inhibited by 4DAMP. Antagonists toward M1, M2, and M4 receptors all failed to alter the current.</p><p><b>CONCLUSION</b>Choline modulates the cellular electrical properties of the heart, probably by activating a K+ current via stimulation of the M3 receptor. M3-R/IK(M3) may act as a new target for antiarrhythmic agents.</p>

Establishment of a novel arrhythmia model in rats / 药学学报

<p><b>AIM</b>To establish a novel arrhythmia model in rats.</p><p><b>METHODS</b>Coronary artery occlusion was produced in hyperlipidemic rats after the animals were fed a high fat and cholesterol chow for 15 days. The incidence, duration and score of arrhythmias were determined 1 hour after coronary occlusion.</p><p><b>RESULTS</b>The incidence, duration and score of arrhythmia induced by coronary artery occlusion increased significantly in hyperlipidemic rats compared with those in normal rats (P < 0.05). In normal rats, pretreatment with amiodarone 60 mg x kg(-1) or verapamil 25 mg x kg(-1) 3 days before coronary artery occlusion did not influence the incidence, duration and score of arrhythmia (P > 0.05). In hyperlipidemic rats, amiodarone 60 mg x kg(-1) decreased the incidence, duration and score of arrhythmia (P < 0.05), but not verapamil 25 mg x kg(-1) (P > 0.05).</p><p><b>CONCLUSION</b>The novel arrhythmia model induced by coronary artery occlusion in hyperlipidemic rats is reliable and similar to the pathophysiological state in human being.</p>

Inhibitory effects of chloride channel blockers NPPB on proliferation of human glomerular mesangial cells / 药学学报

<p><b>AIM</b>To investigate the effects of NPPB, a chloride channel blocker, on proliferation of mesangial cells.</p><p><b>METHODS</b>Cell proliferation was determined by measuring cell number and 3H-thymidine incorporation. The LDH activity released from these cells was measured as evaluation of cell viability. The phase of cell cycle was detected by flow cytometry.</p><p><b>RESULTS</b>Cell proliferation assays showed that treatment with both NPPB (50 and 25 micromol x L(-1)) and in hypertonic media (100% increased osmolarity with D-mannitol ) significantly reduced the number of human MC and 3H-thymidine incorporation in a dose-dependent manner. But the LDH activity was not significantly altered in the treatment with 50 micromol x L(-1) NPPB. Flow cytometry experiments showed that 50 and 25 micromol x L(-1) NPPB arrested (84.2 +/- 2.4) % and (80.8 +/- 2.9) % of cells at G0/G1 stage, versus (70.5 +/- 1.4) % of control cells. Conclusion NPPB suppresses cell proliferation and produces growth arrest at G0/G1 phase in human MC by a mechanism probably associated with changes in cell volume.</p>

Construction and application of prokaryotic expression system of Leptospira interrogans lipL32/1-lipL41/1 fusion gene / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct lipL32/1-lipL41/1 fusion gene and its prokaryotic expression system and to determine frequencies of carrying and expression of lipL32 and lipL41 genes in L.interrogans wild strains and specific antibody levels in sera from leptospirosis patients.</p><p><b>METHODS</b>lipL32/1-lipL41/1 fusion gene was constructed using linking primer PCR method and the prokaryotic expression system of the fusion gene done with routine techniques. SDS-PAGE was used to examine expression of the target recombinant protein rLipL32/1-rLipL41/1. Immunogenicity of rLipL32/1-rLipL41/1 was identified by Western blot. PCR and MAT were performed to detect carrying and expression of lipL32 and lipL41 genes in 97 wild L.interrogans strains. Antibodies against products of lipL32 and lipL41 genes in serum samples from 228 leptospirosis patients were detected by ELISA method.</p><p><b>RESULTS</b>The homogeneity of nucleotide and putative amino acid sequence of lipL32/1-lipL41/1 fusion gene were 99.9 % and 99.8 % in comparison with the reported sequences. Expression output of the target recombinant protein rLipL32/1-rLipL41/1, mainly present in inclusion body, accounted for 10 % of the total bacterial proteins. Both the rabbit antisera against rLipL32/1 and rLipL41/1 could combine to rLipL32/1-rLipL41/1. 97.9 % and 87.6 % of the L.interrogans wild strains had lipL32 and lipL41 genes, respectively. 95.9 % and 84.5 % of the wild strains were positive for MAT with titers of 1:4 - 1:128 using rabbit anti-rLipL32s or anti-rLipL41s sera, respectively. 94.7 % - 97.4 % of the patients'serum samples were positive for rLipL32s antibodies, while 78.5 % - 84.6 % of them were rLipL41s antibodies detectable.</p><p><b>CONCLUSION</b>lipL32/1-jlipL41/1 fusion gene and its prokaryotic expression system were successfully constructed. The expressed fusion protein had qualified immunogenicity. Both the lipL32 and lipL41 genes are extensively carried and frequently expressed by different serogroups of L.interrogans, and their expression products exhibit cross-antigenicity.</p>