Clinical characteristics and risk factors of pericardial effusion after hematopoietic stem cell transplantation in children with thalassemia major / 中华儿科杂志

Objective: To investigate the characteristics, risk factors and outcomes of thalassemia major (TM) children with pericardial effusion (PE) after allo-geneic hematopoietic stem cell transplantation (allo-HSCT). Methods: Clinical data of 446 TM children received allo-HSCT at Shenzhen Children's Hospital between January 2012 and December 2020 were analyzed retrospectively. Patients were divided into PE and non-PE group according to the occurrence of PE. Chi-square tests were used to investigate the risk factors that were associated with the development of PE. Kaplan-Meier method was used for survival analysis of the 2 groups. Results: Twenty-five out of 446 patients (5.6%) developed PE at a time of 75.0 (66.5, 112.5) days after allo-HSCT. Among these patients, 22 cases (88.0%) had PE within 6 months after allo-HSCT and 19 patients (76.0%) had PE within 100 days after allo-HSCT. The diagnoses of PE were confirmed using echocardiography. Pericardial tamponade was observed in only 1 patient, who later undergone emergency pericardiocentesis. The rest of patients received conservative managements alone. PE disappeared in all patients after treatment. Risk factors that were associated with the development of PE after allo-HSCT included the gender of patients, the type of transplantation, the number of mononuclear cells (MNC) infuse, pulmonary infection after HSCT and transplantation associated thrombotic microangiopathy (TA-TMA) (χ²=3.99, 10.20, 14.18, 36.24, 15.03, all P<0.05). In 239 patients that received haploidentical HSCT, the development of PE was associated with the gender of patients, pulmonary infection after HSCT and TA-TMA (χ²=4.48, 20.89, 12.70, all P<0.05). The overall survival rates of PE and non-PE groups were 96.0% (24/25) and 98.6% (415/421). The development of PE was not associated with the overall survival of TM children after allo-HSCT (χ²=1.73, P=0.188). Conclusions: PE mainly develop within 100 days after allo-HSCT in pediatric TM recipients. Haploidentical grafts, female gender, pulmonary infection after HSCT and TA-TMA are the main risk factors associated with PE development after transplant. However, the presence of PE don't have a significant impact on the outcomes of pediatric TM patients after allo-HSCT.

Effect of Addition and Subtraction Adjuvant Therapy of Bufeitang on Immune Inflammation After Tracheotomy in Stroke Patients with Syndrome of Deficiency of Spleen and Lung Qi / 中国实验方剂学杂志

Objective:To discuss the efficacy of addition and subtraction adjuvant therapy of Bufei decoction for pulmonary infection after tracheotomy in stroke patients (syndrome of deficiency of spleen and lung Qi) and investigate its effect on immune inflammation. Method:One hundred patients were randomly divided into control group (50 cases) and observation group (50 cases) by random number table. The patients in both groups got cefepime hydrochloride for injection, once every 12 hours, 2 g/time， at the same time, symptomatic and supportive comprehensive treatment was given. Patients in control group additionally got compound glycyrrhiza oral solution via gastric tube, 10 mL/time, 3 times/day. Patients in observation group got addition and subtraction adjuvant therapy of Bufeitang every morning and night via gastric tube, 1 dose/day. The treatment course was 14 days in both groups. At the 1st, 7th and 14th day after treatment, scores of clinical pulmonary infection scale (CPIS) and Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) were graded. The time to control pulmonary infection and the antibiotics use time were recorded. Before and after treatment, levels of T lymphocyte subsets (CD3+, CD4+，CD8+ and CD4+/CD8+), regulatory T cells of (Treg cells), immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M(IgM), procalcitonin (PCT), tumor necrosis factor-α (TNF-α), interleukin-1β, IL-6 and IL-10 were detected, and safety was evaluated. Result:At the 7th and 14th day after treatment, scores of CPIS and APACHE Ⅱ in observation group were lower than those in control group (P<0.01). The time to control pulmonary infection and antibiotics use time were shorter than those in control group (P<0.01). Levels of Treg cells, CD4+ and CD4+/CD8+ were higher than those in control group (P<0.05). Levels of CD8+, PCT, TNF-α, IL-1β, IL-6 and IL-10 were lower than that in control group (P<0.01), while levels of IgA and IgM were higher than those in control group (P<0.01). There was no adverse reaction related to Bufeitang. Conclusion:Based on comprehensive treatment of western medicine for anti-infection and symptomatic support, addition and subtraction adjuvant therapy of Bufeitang can effectively control the severity of pulmonary infection caused by tracheotomy in stroke, reduce coughing and expectoration, shorten the course of pulmonary infection and the use time of antibiotics, regulate immune function and inhibit inflammatory reaction.

Gingerol activates noxious cold ion channel TRPA1 in gastrointestinal tract / 中国天然药物

TRPA1 channels are non-selective cation channels that could be activated by plant-derived pungent products, including gingerol, a main active constituent of ginger. Ginger could improve the digestive function; however whether ginger improves the digestive function through activating TRPA1 receptor in gastrointestinal tract has not been investigated. In the present study, gingerol was used to stimulate cell lines (RIN14B or STC-1) while depletion of extracellular calcium. TRPA1 inhibitor (rethenium red) and TRPA1 gene silencing via TRPA1-specific siRNA were also used for mechanistic studies. The intracellular calcium and secretion of serotonin or cholecystokinin were measured by fura-2/AM and ELISA. Stimulation of those cells with gingerol increased intracellular calcium levels and the serotonin or cholecystokinin secretion. The gingerol-induced intracellular calcium increase and secretion (serotonin or cholecystokinin) release were completely blocked by ruthenium red, EGTA, and TRPA1-specific siRNA. In summary, our results suggested that gingerol derived from ginger might improve the digestive function through secretion releasing from endocrine cells of the gut by inducing TRPA1-mediated calcium influx.

Combination therapy with pegylated interferon alpha-2b and adefovir dipivoxil in HBeAg-positive chronic hepatitis B versus interferon alone: a prospective, randomized study / 华中科技大学学报（医学）（英德文版）

Currently available monotherapies of oral nucleoside/nucleotide analogs or interferon are unable to achieve a sustained and effective response in most of patients with chronic hepatitis B (CHB). The objective of the present study was to compare the efficacy and safety of pegylated interferon (Peg-IFN) alpha-2b plus adefovir dipivoxil combination therapy versus Peg-IFN alpha-2b alone. Sixty-one HBeAg-positive chronic hepatitis B patients were randomized to receive Peg-IFN alpha-2b alone (1.5 μg/kg once weekly) or Peg-IFN alpha-2b plus adefovir (10 mg daily) for up to 52 weeks. Efficacy and safety analyses were performed on all participants who received at least one dose of study medication. The rate of HBeAg seroconversion and undetectable HBV-DNA were evaluated after 52 weeks of therapy. At the end of treatment, 11 of 30 (36.7%) patients receiving combination therapy achieved HBeAg seroconversion versus 8 of 31 (25.8%) in the monotherapy group (P=0.36). In contrast, the percentage of patients with undetectable serum HBV DNA was significantly higher in the combination group than in the monotherapy group (76.7% vs. 29.0%, P<0.001). Thyroid dysfunction was more frequent in the combination group than in the monotherapy group (P<0.05). In HBeAg-positive CHB, combination of Peg-IFN alpha-2b and adefovir for 52 weeks resulted, at the end of treatment, in a higher virological response but without significant impact on the rate of HBeAg seroconversion and possibly an adverse effect on thyroid function.

Combination therapy with pegylated interferon alpha-2b and adefovir dipivoxil in HBeAg-positive chronic hepatitis B versus interferon alone: a prospective, randomized study / 华中科技大学学报（医学）（英德文版）

Currently available monotherapies of oral nucleoside/nucleotide analogs or interferon are unable to achieve a sustained and effective response in most of patients with chronic hepatitis B (CHB). The objective of the present study was to compare the efficacy and safety of pegylated interferon (Peg-IFN) alpha-2b plus adefovir dipivoxil combination therapy versus Peg-IFN alpha-2b alone. Sixty-one HBeAg-positive chronic hepatitis B patients were randomized to receive Peg-IFN alpha-2b alone (1.5 μg/kg once weekly) or Peg-IFN alpha-2b plus adefovir (10 mg daily) for up to 52 weeks. Efficacy and safety analyses were performed on all participants who received at least one dose of study medication. The rate of HBeAg seroconversion and undetectable HBV-DNA were evaluated after 52 weeks of therapy. At the end of treatment, 11 of 30 (36.7%) patients receiving combination therapy achieved HBeAg seroconversion versus 8 of 31 (25.8%) in the monotherapy group (P=0.36). In contrast, the percentage of patients with undetectable serum HBV DNA was significantly higher in the combination group than in the monotherapy group (76.7% vs. 29.0%, P<0.001). Thyroid dysfunction was more frequent in the combination group than in the monotherapy group (P<0.05). In HBeAg-positive CHB, combination of Peg-IFN alpha-2b and adefovir for 52 weeks resulted, at the end of treatment, in a higher virological response but without significant impact on the rate of HBeAg seroconversion and possibly an adverse effect on thyroid function.

Effects of scutellaria stem-leaf total flavonoids on cardiocyte apoptosis induced by hypoxia/reoxygenation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of Scutellaria baicalensis stem-leaf total flavonoids (SSTF) on cardiocyte apoptosis of neonatal rats induced by hypoxia/reoxygenation and its action of mechanism. METHODS Sixty one to two days old rats, male or female, were selected. Hypoxia/reoxygenation injured model was established in cultured cardiocytes of neonate rats. The cultured neonatal rat cardiocytes were divided into 5 groups, i.e., the normal control group, the hypoxia/reoxygenation injury group (as the model group, cultured cardiocytes were exposed to hypoxia 2 h and subsequently reoxygenated for 4 h), the hypoxia/reoxygenation injury plus 50 mg/L SSTF group (as the low dose SSTF group), the hypoxia/reoxygenation plus 100 mg/L SSTF group (as the middle dose SSTF group), and the hypoxia/reoxygenation plus 200 mg/L SSTF group (as the high dose SSTF group). The cell viability was detected by methyl thiazolyl tetrazolium (MTT) colorimetry. The apoptosis of cardiocytes was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and the apoptosis rate calculated. The Bcl-2 and Bax protein expressions were determined by immunohistochemistry.</p><p><b>RESULTS</b>Compared with the normal control group, the cell viability, Bcl-2 protein contents and Bcl-2/Bax decreased, the apoptosis rate and Bax protein contents increased in the model group (all P<0.01). Compared with the model group, the cell viability, Bcl-2 protein contents and Bcl-2/Bax increased, while the apoptosis rate and Bax protein contents decreased in each SSTF treated group (P<0.05, P<0.01). Compared with the low dose SSTF group, significant difference existed in each index of the high dose SSTF group (all P<0.05).</p><p><b>CONCLUSIONS</b>SSTF had protection on hypoxia/reoxygenation induced cardiocyte apoptosis. Its protective mechanism might be correlated with its up-regulation of the expression of Bcl-2 protein ahd down-regulation of the expression of Bax protein.</p>

Association of the neprilysin and apolipoprotein E gene polymorphisms with Alzheimer's disease in Xinjiang Uygur population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association between the neprilysin (NEP) gene and apolipoprotein E (ApoE) gene polymorphisms and sporadic Alzheimer's disease (SAD) in Xinjiang Uygur population.</p><p><b>METHODS</b>The polymorphisms of the NEP and ApoE gene were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 111 SAD patients and 117 healthy controls.</p><p><b>RESULTS</b>(1) The frequency of the T allele in the NEP gene was significantly higher in the SAD patients than that in the controls (Chi-square= 5.005, P< 0.05); and there was higher risk to develop SAD in the T allele carriers. (2) The frequency of the ApoE 4 epsilon 4 allele was higher in the SAD patients than in the controls (Chi-square= 4.218, P< 0.05); and the ApoE 4 epsilon 4 carriers had significantly increased risk of developing SAD. (3) No significant interaction was found between the NEP and ApoE polymorphisms in SAD patients.</p><p><b>CONCLUSION</b>The NEP and ApoE gene polymorphisms may be associated with SAD. NEP gene may be an independent genetic risk factor for SAD in Xinjiang Uygur population.</p>

Calorie control increased vaspin levels of serum and periepididymal adipose tissue in diet-induced obese rats in association with serum free fatty acid and tumor necrosis factor alpha / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Vaspin was recently identified as a novel adipokine that is predominantly secreted from adipose tissue and exerts insulin-sensitizing effects. This study was undertaken to elucidate the regulative effects of calorie control on the expression of vaspin and its potential mechanism.</p><p><b>METHODS</b>Diet-induced obese Sprague Dawley (SD) rats were adopted as experimental models and accepted interventions of various ingestions and pioglitazone. Various differentiated stages of cultured 3T3-L1 cells were dealt with pioglitazone or TNFalpha in vitro for 48 hours to further verify findings in animal experiments.</p><p><b>RESULTS</b>The rats were successfully induced into an obese experimental model with hyperinsulinemia, hyperlipidemia, and increased serum free fatty acid and TNFalpha by 12-week high-fat diet. It was found that depending on whether the rats were fed by a high-fat diet or a basal diet, there was extremely higher vaspin in the periepididymal fat pad than in subcutaneous adipose tissues by 16 weeks. Vaspin in sera and the periepididymal fat pad was much lower in rats with a high-fat diet than those with a basal diet (all P < 0.05), but vaspin in subcutaneous fat tissues was prone to increase in rats with a high-fat diet. A 4-week calorie restriction or pioglitazone on the obese rats resulted in a partial recovery of vaspin levels in sera and periepididymal adipose tissues, especially the latter revealed a more obvious superiority and increased vaspin levels of subcutaneous adipose. Surprisingly, the treatment of 4-week high-fat diet on non-obese rats did not significantly depress vaspin of sera and periepididymal adipose tissues. However, it is unknown if re-feeding generated the effect on vaspin levels of obese and non-obese rats on sera or adipose tissues. The correlation analysis showed that vaspin levels of serum and periepididymal fat tissues were negatively correlated with serum FFA, TNFalpha and insulin; meanwhile, there was a positive correlation between serum vaspin and vaspin of periepididymal fat tissues. Pioglitazone enhanced vaspin levels in cultured 3T3-L1 cells and supernatant in various differentiated stages, and this effect became more and more obvious along with the change of preadipocytes into mature fat cells. Administration of TNFalpha caused suppression on vaspin expression in differentiated stages of 3T3-L1 cells.</p><p><b>CONCLUSIONS</b>The present data indicated that a long-term high-fat diet could induce obesity metabolic syndrome in SD rats and finally lead to lower vaspin of sera and periepididymal fat, while pioglitazone and chronic calorie-control ingestion could enhance the production of vaspin. It was undoubtedly demonstrated that vaspin expression was strongly associated with insulin sensitivity, serum FFA, and TNFalpha.</p>

Controlled study of auricular point taping and pressing therapy for treatment of vascular dementia / 中国针灸

<p><b>OBJECTIVE</b>To assess the therapeutic effect of auricular point taping and pressing therapy on vascular dementia (VD).</p><p><b>METHODS</b>One hundred and eighty cases of vascular dementia were randomly divided into an auricular point taping and pressing group and a western medicine group, 90 cases in each group. The auricular point taping and pressing group was treated by auricular point taping and pressing at auricular points Shennen, Nao (brain), Shen (kidney), Zhen (pulvinal), and the western medicine group by oral administration of Nimodipine, thrice each day, 30 mg each time. They were treated for 12 weeks. The scores of mini-mental state examination (MMSE) and activities of daily living (ADL), and adverse reaction were observed.</p><p><b>RESULTS</b>The scores of MMSE before treatment and 12 weeks after treatment were 18.00 +/- 3.88 and 20.83 +/- 3.74 in the auricular point taping and pressing group, and 17.80 +/- 3.82 and 20.70 +/- 3.53 in the western medicine group, respectively, with significant increases in scores of MMSE after treatment in the two groups (both P < 0.01) and with no significant difference between the two groups (P > 0.05). The scores of ADL before treatment and 12 weeks after treatment were 44.90 +/- 14.84 and 39.60 +/- 12.45 in the auricular point taping and pressing group, and 45.70 +/- 14.86 and 39.10 +/- 13.44 in the western medicine group, respectively, with significant decreases after treatment in the two groups (both P < 0.01) and with no significant difference between the two groups (P > 0.05). In the auricular point taping and pressing group, 2 cases withdrew from the test because adhesive plaster allergic reaction induced severe itch of the auricle. In the western medicine group, one case had mild dizziness and another case had diarrhea respectively.</p><p><b>CONCLUSION</b>Auricular point taping and pressing and Nimodipine have similar therapeutic effect on vascular dementia, and have no obvious adverse reaction, except adhesive plaster allergic reaction.</p>

A novel mutation of 428delG in DAX-1 gene causing X-linked adrenal congenital hypoplasia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the clinical features and to identify the DAX-1 gene mutation in a Chinese kindred with X-linked adrenal hypoplasia congenital(AHC).</p><p><b>METHODS</b>Clinical data and peripheral blood samples were obtained from the affected individuals and their relatives. The genomic DNA was isolated from whole blood. Four pairs of primers were used to amplify the two exons of the DAX-1 gene, and PCR products were purified and sequenced directly. Sequencing results were compared to the human DAX-1 sequence in the public database.</p><p><b>RESULTS</b>A novel hemizygous frameshift mutation (428delG) in exon 1 of the DAX-1 gene was found in both patients (the index case and his cousin). Some clinical features such as the age of onset were different although these 2 patients carried the same mutation. Three females in the family, including the mothers of the 2 patients and their grandmother were carriers of this mutation. No such mutation was detected in other healthy persons in the family.</p><p><b>CONCLUSION</b>The result suggested that X-linked AHC in the kindred was caused by a novel mutation of 428delG in the DAX-1 gene, and the same mutation can give rise to variable phenotypes.</p>

Survey on central obesity among middle-aged and elderly people of Uygurs and Hans ethnicities in Xinjiang / 中华流行病学杂志

Objective To study central obesity among middle-aged and elderly residents of Xinjiang Uygur and Han ethnicities, living in rural and urban areas. Methods Multi-stage stratified random cluster sampling approaches were adopted to collect data from 6 areas in Soutbem, Eastern, Northern Xinjiang and Urumqi city community, from July of 2005 to June of 2007. Results 8284 people were investigated to have found that the crude prevalence rate and the adjusted standardized incidence were 50.11% and 55.40% respectively, on central obesity. The figures were higher than the national level, according to the 2000 census age composition of Xinjiang. The prevalence rate of central obesity was higher in males than in females (P<0.05) higher in residents of Uygur than in Han ethnicities(P<0.05). The prevalent rates of the central obesity hypertension, diabetes and dyslipidemia were higher than those of non-obese ones (P<0.05). Conclusion The standardized prevalence rates of central obesity in residents with Xinjiang Uygur and Hart ethnicities were higher than data from the national statistics. Differences were found in ethnicity, gender and age. The prevalence rates of hypertension, diabetes and dyslipidemia in people having central obesity were higher than the non-obese ones.

Association of the low density lipoprotein receptor-related protein gene 766C/T polymorphism with Alzheimer's disease in Xinjiang Uygurs and Hans / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the low density lipoprotein receptor-related protein gene (LRP) 766C/T polymorphisms and Alzheimer's disease (AD) in Xinjiang Uygurs and Hans populations.</p><p><b>METHODS</b>Those included in the study were > or = 50 years of age and of either Xinjiang Uygur or Han descents. Two hundred and nine individuals had AD and 220 were healthy controls. They were recruited according to ADRDA-NINCDS criteriaîThe polymorphisms of the LRP gene were determined by the PCR-restriction fragment length polymorphism technique. The case-control analysis was adopted to analyze the frequencies of genotypes and alleles.</p><p><b>RESULTS</b>(1) The distribution of genotypes or alleles of LRP gene had significant differences between the AD group and the control group in both the Xinjiang Uygurs and Hans populations (P < 0.05). (2) The frequencies of genotypes and alleles were significantly different between the Han AD and Han control group (P < 0.05). (3) The frequencies of genotypes and alleles in those > or = 65 years were significantly different from that in others (P < 0.05). There was a significant increase of AD in the C allele carriers (OR=1.98, P < 0.05). (4) The frequencies of the CC genotype and C allele in female AD patients were higher than that in female controls (P < 0.05), and the C allele carriers had significant increase of AD (OR=2.927, P < 0.05).</p><p><b>CONCLUSION</b>The LRP 766C/T polymorphisms were significantly different between the Chinese Xinjiang Uygur and Han populations. The LRP 766C/T polymorphisms might be associated with AD in the Han population, in females and those of > or = 65 years old.</p>

Construction of prokaryotic expression system of Salmonella paratyphi A spaO gene and immunogenicity and immunoprotection of the expressed product / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the immunogenicity and immunoprotection of the recombinant expressing product (rSpaO) of S. paratyphi A spaO gene, and to demonstrate the frequencies of spaO gene carrying and expressing in S. paratyphi A isolates.</p><p><b>METHODS</b>The spaO gene of a clinical S. paratyphi A strain JH01 was amplified and then cloned. After sequencing of the cloned spaO gene, a prokaryotic expression system of the gene was constructed. SDS-PAGE were applied to examine the rSpaO expression. Ni-NTA affinity chromatography was performed to collect rSpaO. Immunogenicity of rSpaO was determined by Western blot assay. A PCR assay and an ELISA were established to respectively detect the carrying and expressing frequencies of the spaO genes in 98 S. paratyphi A isolates. The immunoprotective effects of rSpaO in S. paratyphi A strain 50001 infected mice were observed.</p><p><b>RESULTS</b>In comparison with the reported corresponding sequences, the nucleotide and putative amino acid sequence homologies of the cloned spaO gene were 99.45%-99.89% and 99.01%-100%, respectively. The expression output of rSpaO was approximately 75% of the total bacterial proteins. S. paratyphi A antiserum could recognize as well as combine with rSpaO. rSpaO could efficiently induce rabbits to produce specific antibody. 94.9% (93/98) of the S. paratyphi A isolates had spaO gene and 91.4% (85/93) of the spaO+ strains could express SpaO. 58.3% and 50.0% of the mice that oral-taken or subcutaneous injected with 500 microg of rSpaO for immunization were survival after challenged by lethal dose of S. paratyphi A strain 50001. When co-immunized with 5 microg rLTB, the survival rates of the mice increased to 88.3% and 75.0%, respectively.</p><p><b>CONCLUSION</b>The S. paratyphi isolates had relatively high carrying and expressing frequencies of spaO gene. rSpaO showed a fine immunogenicity and a certain immunoprotective effect, which could be used as an antigen candidate for developing genetic engineering vaccine of S. paratyphi.</p>

Morphological and molecular biological peculiarities of the experimental autoimmune prostatitis rat model / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the morphological and molecular biological peculiarities of the experimental autoimmune prostatitis (EAP) rat model made by SC purified prostate protein twice with immune adjuvant.</p><p><b>METHODS</b>Male rats were intradermally immunized with a saline extract of male rat prostate glands (RPG) in Freund's complete adjuvant (FCA) and Pertussis-Diphtheria-Tetanus vaccine 0.5 ml i.p. at the 0 and 30th day, and the concentrations of the extract were respectively 5 mg/ml, 10 mg/ml and 15 mg/ml. At the 45th day, the rats were sacrificed and the morphological and molecular biological changes of the prostate specimens were observed to determine the effective concentration of RPG for a successful model.</p><p><b>RESULTS</b>The expression of inflammation genes such as TNF-alpha, IL-1beta, IL-2 and iNOS obviously increased in the high-dosage model group; LM, EM and in situ hybridization revealed appearant chronic inflammation response, but this was not the case in the other two dosage groups.</p><p><b>CONCLUSION</b>15 mg/ml RPG mixed with FCA (1:1) 1.0 ml SC with Pertussis-Diphtheria-Tetanus vaccine 0.5 ml i.p. was an effective dosage for the successful model in our experiment.</p>

Identification on the immunogenic activity of recombinant rLTB/CTB-LipL41/ 1 to Leptospira interrogans and detection of lipL41 gene with its production / 中华流行病学杂志

<p><b>OBJECTIVE</b>To construct prokaryotic expression systems of ltB/ctB-lipL41/1 fusion genes, identify immunogenic and adjuvant activities of the products as well as to understand the frequencies of lipL41 gene that carrying and expressing in L. interrogans wild strains and specific antibody levels in sera from patients with leptospirosis.</p><p><b>METHODS</b>Polymerase chain reaction (PCR) with linking primer was applied to construct the fusion genes ltB-lipL41/1 and ctB-lipL41/1. By routine molecular biological techniques, prokaryotic expression systems of the two fusion genes were constructed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to examine expression of the target recombinant proteins rLTB-rLipL41/1 and rCTB-rLipL41/1. Both western blot and Ganglioside-enzyme linked immunosorbent assay (GM-ELISA) were used while immunogenic and adjuvant activities of rLTB-rLipL41/1 and rCTB-rLipL41/1 were measured. PCR and MAT were performed to detect lipL41 gene and expression of the gene in 97 wild strains of L. interrogans, respectively. Antibodies against product of lipL41 gene in serum samples from 228 leptospirosis patients were detected by ELISA.</p><p><b>RESULTS</b>In comparison with reported corresponding sequences, the similarities of ltB-lipL41/1 and ctB-lipL41/1 fusion genes to nucleotide and putative amino acid sequence were 99.6%-99.9% and 99.8%-100%, respectively. Expression outputs of both rLTB-rLipL41/1 and rCTB-rLipL41/1, mainly presenting with inclusion body, consisting approximate 10% of the total bacterial proteins. Both rLTB-rLipL41/1 and rCTB-rLipL41/1 could combine rabbit anti-rLipL41/1 serum as well as bovine GM1, respectively. 87.6% of the L. interrogans wild strains(85/97) having lipL41 gene while 84.5% (82/97) of the wild strains with rLipL41/1 or rLipL41/2 antiserum were positive for MAT with titers of 1:4-1:128. 84.6% (193/ 228), 78.5% (179/228) from the patients' serum samples were positive for rLipL41/1 and rLipL41/2 antibodies, respectively.</p><p><b>CONCLUSION</b>ltB-lipL41/1 and ctB-lipL41/1 fusion genes and their prokaryotic expression systems were successfully constructed in this study. The two expressed fusion proteins showed qualified immunogenic and adjuvant activities. lipL41 gene was extensively distributed and frequently expressed in different serogroups of L. interrogans. rLTB-rLipL41/1 or rCTB-rLipL41/1 seemed to have had good potential to serve as an antigen in L. interrogans genus-specific vaccine.</p>