In order to amplify alpha toxin gene of Clostridium septicum HeB01 strain, one pair of primers was designed according to the GenBank sequence, and a 1323bp alpha toxin gene fragment was obstained by PCR. Sequence analysis indicated that the homology of the nucleotid sequence of HeB01 strain to those other reference strains was more than 99.5% . The expression plasmid pQE30-alpha was constructed by inserting alpha toxin gene into the prokaryotic expression vector pQE30. The plasmid expressed when the recombinant strain M15(pQE30-alpha) was induced by IPTG. The specific 48 kD protein was detected SDS-PAGE and the immunogenicity of the expressed alpha toxin was confirmed by Western blot and ELISA. The expressed alpha toxin was transformed into alpha toxoid vaccine by adding 0.3% formaldehyde into alpha toxin. The protective immune response was proved after the mice was immunized with alpha toxoid vaccine. The results showed that the recombinanted strain M15 (pQE30-alpha) could be as a candidate of alpha toxoid vaccine to provide protective immune response against clostridium septicum infection.
Animals ; Antibodies, Bacterial ; blood ; immunology ; Bacterial Toxins ; genetics ; immunology ; metabolism ; Blotting, Western ; Cloning, Molecular ; Clostridium Infections ; immunology ; microbiology ; prevention & control ; Clostridium septicum ; genetics ; immunology ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Gene Expression Regulation, Bacterial ; Mice ; Polymerase Chain Reaction ; Recombinant Proteins ; administration & dosage ; immunology ; metabolism ; Toxoids ; immunology ; Vaccination

Purpose To examine the expression of Fascin-1 and β-catenin protein and K-ras gene mutation in colorectal adenocarcinoma, and to explore their role in progression of colorectal neoplasm and their relevance. Methods Fascin-1 and β-catenin were analyzed by use of immunohistochemistry En Vision two-step. K-ras gene mutation was detected by ARMS method.Relationship between overexpression of Fascin-1, the nuclear expression of β-catenin, and the mutations of K-ras gene and clinicopathologic parameters was analyzed, the correlation between them was also analyzed. Results In 112 colorectal adenocarcinoma samples, the overexpression rate of Fascin-1 protein was 27.7％ (31/112), significantly higher than non-neoplastic mucosa (P < 0.01). The high nuclear expression rate of β-catenin was 29.5％ (33/112) in adenocarcinoma and non-neoplastic mucosa respectively with a significant difference between two groups (P < 0.01). High expression rate of Fascin-1 protein and β-catenin were correlated significantly with lymph node metastasis (P = 0.022, P = 0.027), and TNM staging (P =0.042, P = 0.019) in colorectal adenocarcinoma. The overexpression of Fascin-1 protein was correlated with tumor location (P = 0.004). The mutation rate of K-ras gene was 34.8％ (39/112), which showed no correlation with age, gender, tumor size, grade of differentiation, lymph node metastasis and TNM staging (P> 0.05). There was a correlation between the overexpressison of Fascin-1 protein, the nuclear expression of β-catenin and the mutation of K-ras gene (rs= 0.252, rs= 0.258, P < 0.05). The overexpression of Fascin-1 protein positively correlated with the nuclear expression of β-catenin (rs= 0.213, P < 0.05). Conclusion Fascin-1 protein and β-catenin protein are involved in invasion and metastasis of colorectal cancer and are associated with K-ras gene mutation. K-ras may promote the overexpression of Fascin-1 by virtue of nuclear expression ofβ-catenin, which provided a new research direction on the treatment of K-ras gene mutated colorectal adenocarcinoma.

0.05). The RBC folate level of birth defect group except the urinary defect was significantly lower compared with the control group(233-547 vs 689 nmol/L,P

Objective To evaluate the efficacy of volume therapy guided by stroke volume variabil-ity ( SVV) in the patients undergoing surgery for severe traumatic brain injury. Methods Thirty patients of both sexes with severe traumatic brain injury, aged 18-64 yr, of American Society of Anesthesiologists physical status Ⅲ, who were admitted to the hospital within 24 h after injury, with Glasgow Coma Scale ( GCS) score≤8, were divided into control group ( C group, n=15) and SVV group ( n=15) using a ran-dom number table method. In group C, conventional fluid administration was performed to maintain mean arterial pressure at 65-110 mmHg, central venous pressure at 5-12 cmH2 O and urine volume>1 ml·kg-1 ·h-1 . Fluid was given according to SVV, maintaining SVV≤13％ and mean arterial pressure at 65-110 mmHg in group SVV. Immediately after skin incision ( T0 ) , immediately after opening cerebral dura mater ( T1 ) , at 1 h after opening cerebral dura mater ( T2 ) , immediately after suturing cerebral dura mater ( T3 ) and at the end of operation ( T4 ) , blood samples were collected from the radial artery and inter-nal jugular venous bulb for blood gas analysis, the jugular venous oxygen partial pressure, jugular venous bulb oxygen saturation, blood lactate, arterial oxygen partial pressure, arterial oxygen saturation and Hbwere recorded, and the cerebral artery and arteriovenous blood O2 content difference and cerebral O2 extrac-tion rate were calculated. Blood samples were collected from the internal jugular venous bulb at T0-2 , T4 and 24 h after operation ( T5 ) for determination of S100β protein concentrations by enzyme-linked immunosor-bent assay. The intraoperative volume of fluid intake and output and consumption of vasoactive drugs were recorded. GCS scores were recorded immediately after admission to the operating room, and at 1, 3, 7 and 14 days after operation. The development of postoperative length of hospitalization and complications ( pul-monary infection and brain edema) was recorded. Glasgow Outcome Scale Score was used to assess the early postoperative quality of life. Results Compared with group C, the urine volume was significantly in-creased, the consumption of vasoactive drugs was reduced, jugular venous bulb oxygen saturation was in-creased at T2,3 , the cerebral O2 extraction rate was decreased at T2-4 , the serum S100β protein concentra-tion was decreased at T2 , and the GCS score was increased at day 3 after operation ( P<0. 05) , and no sig-nificant change was found in blood lactate, postoperative Glasgow Outcome Scale score or length of hospital-ization at each time point in group SVV ( P>0. 05) . Conclusion SVV-guided volume therapy can improve cerebral oxygen metabolism, ensure adequate tissue perfusion and reduce craniocerebral injury in the pa-tients undergoing surgery for severe traumatic brain injury.

OBJECTIVETo observe the effect of Tongsaimai (TSM) tablets in treating foot trauma of diabetic foot (DF) model rats, and discuss its potential mechanism.

METHODMale SD rats were selected to duplicate the diabetic foot ulcer model and randomly divided into the blank control group, the model group, the metformin treatment group, and TSM 12.44, 6.22, 3.11 g x kg(-1) groups (n = 10). The healing of ulcer wounds were observed on day 1, 4, 8, 13 and 18. After 18 days, a histopathologic examination was conducted for ulcer tissues. The contents of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by hydroxylamine and TBA methods. The content of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were determined with the radioimmunoassay. The immunohistochemical method was used to observe the expression of vascular endothelial growth factor (VEGF) in ulcer tissues and the number of capillary vessels.

RESULTTSM could alleviate the pathological changes of diabetic foot rats, accelerate the ulcer healing on 4, 8, 13, 18 d, reduce MDA, IL-6, TNF-alpha, VEGF content in rat serum at 18 d (after the rehabilitation period), and enhance the SOD content. Specifically, the TSM 12.44 g x kg(-1) group showed significant differences compared with the model group (P < 0.05, P < 0.01). At 18 d after the treatment (the late rehabilitation period), the VEGF expression of TSM 12.44, 6.22 g x kg(-1) groups and the number of blood capillaries of the TSM 12.44 g x kg(-1) group were significantly lower than that of the model group (P < 0.05, P < 0.01).

CONCLUSIONTSM could promote the foot wound healing of DF model rats, reduce MDA, IL-6 and TNF-alpha levels in serum, increase the SOD content and decrease the VEGF expression and the number of blood capillaries in the late rehabilitation period. Its action mechanism may be related to the inhibition of oxidative stress injury and the inflammatory cell infiltration.

Animals ; Diabetic Foot ; drug therapy ; genetics ; metabolism ; physiopathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Interleukin-6 ; genetics ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; genetics ; metabolism ; Tablets ; administration & dosage ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Wound Healing ; drug effects

Objective: To explore the association between chronic hepatitis B virus infection and diabetes among adults. Methods: The baseline data of China Kadoorie Biobank ( CKB ) study from Tongxiang of Zhejiang Province was used for analysis. Community residents were investigated in the study from August 2004 to May 2008, including questionnaire survey, physical measurement and biological sample test. Univariate and multivariate logistic regression models were used to estimate the association of chronic hepatitis B virus infection with diabetes. Results: Totally 52 888 participants were included in the final analysis. The overall prevalence of HBsAg-positive was 3.55% ( N=1 877 ). The overall prevalence of diabetes was 5.17% ( N=2 733 ). The prevalence of HBsAg-positive in diabetic patients was 3.51% ( N=96 ). Both univariate and multivariate logistic regression models indicated that there was no association between chronic hepatitis B virus infection and diabetes（ P>0.05 ）. Conclusion No significant association has been found between chronic hepatitis B virus infection and diabetes among adults.

Objective To understand the epidemiological characteristics of age distribution of measles and related policies on measles vaccines (live; MV) in infants through analyzing the antibody levels of comparison in maternal-infant pairs. Transition of immunity in infants was also studied to provide theoretic basis for measles immunization strategy and to reduce the incidence of month-old infants. Methods In cities of Ningbo, Harbin, and Jinan from Zhejiang, Heilongiiang and Shandong provinces, data was collected from 2004 to 2007 and analyzed regarding the epidemic situation of measles. Studies on maternal-transferred measles antibody were carried our sero-epidemiologically. Results Most of the measles cases were found among babies younger than 12 months,and the incidence of < 1 year olds had been increasing.The distribution was dominated by 5-8 month olds in infant measles cases. The positive rate and GRMT of measles antibody in newborns were 89.3 percent and 738.93. The positive rate of the measles antibody and GMRT of the 6-month infant were 6.9 % and 6.89, while 6.7 % and 3.69 in 8-month infant. There wasa declining trend of the positive rate of the measles antibody during the newborns to 8-month infant. The positive rate and GRMT of measles antibody in mothers were 84.3 percent and 516.94. Mother's measles antibodies mainly to be at low and moderate level, which accounted for 50.4 percent and 30.3 percent respectively, the correlation coefficient between mother and infant was 0. 840. Conclusion Maternal-transferred measles antibody decreased as the growth of infants. The positive rates of measles antibody were quite low in 6-month and 8-month olds which were the age range that needs most attention.

Bismuth modified boron doped diamond (BDD) film electrode was employed for simultaneous determination of trace ZnⅡ,CdⅡand PbⅡby anodic stripping voltammetry.BiⅢwas simultaneously in-situ deposited on bismuth modified boron doped diamond electrode with ZnⅡ,CdⅡ and PbⅡ by pre-concentration.In the presence of BiⅢ,the sensitivity for determination of ZnⅡ,CdⅡ and PbⅡ was remarkably enhanced.Influence factors such as bismuth concentration,boron doped concentrations of BDD electrode,pH,preconcentration potential were investigated and optimized.Under the optimal conditions,the stripping peak currents increased linearly with the increasing concentration of ZnⅡ,CdⅡ and PbⅡ in the range of 10-300 μg/L.The limit of detection was 0.56 μg/L for ZnⅡ,0.32 μg/L for CdⅡand 0.75 μg/L for PbⅡ (S/N=3),respectively.The interference experiments showed that common ions had little influence on the determination except CuⅡ.In addition,the developed electrode displayed a good repeatability.The method was successfully applied to determination of ZnⅡ,CdⅡ and PbⅡ in real water samples with the standard addition recoveries of 92.0%-114.0%.

Endodontic diseases are a kind of chronic infectious oral disease.Common endodontic treatment concepts are based on the removal of inflamed or necrotic pulp tissue and the replacement by gutta-percha.However,it is very essential for endodontic treatment to debride the root canal system and prevent the root canal system from bacterial reinfection after root canal therapy(RCT).Recent research,encompassing bacterial etiology and advanced imaging techniques,contributes to our understanding of the root canal system's anatomy intricacies and the technique sensitivity of RCT.Success in RCT hinges on factors like patients,infection severity,root canal anatomy,and treatment techniques.Therefore,improving disease management is a key issue to combat endodontic diseases and cure periapical lesions.The clinical difficulty assessment system of RCT is established based on patient conditions,tooth conditions,root canal configuration,and root canal needing retreatment,and emphasizes pre-treatment risk assessment for optimal outcomes.The findings suggest that the presence of risk factors may correlate with the challenge of achieving the high standard required for RCT.These insights contribute not only to improve education but also aid practitioners in treatment planning and referral decision-making within the field of endodontics.

OBJECTIVETo assess the effect and safety of Jinhua Qinggan Granule (JHG) in treating influenza patients of wind-heat affecting Fei syndrome (WHAFS).

METHODSTotally 136 influenza patients of WHAFS were randomized by stratification into 3 groups, the high dose JHG group (44 cases, 10 g each time), the low dose JHG group (45 cases, 5 g JHG + 5 g placebo each time), and the placebo control group (47 cases, 10 g placebo each time). All medication was administered three times daily for 5 days. The fever disappearance time, the fever disappearance rate, efficacy of TCM syndrome, the disappearance rate of main symptoms and physical signs of flu, the negative rate of virus nucleic acid in the pharyngeal secretion, and safety indicators were assessed.

RESULTSThe median fever disappearance time was 32.8 h (95% CI: 22.5-41.0 h) in the high dose JHG group, 26.0 h (95% CI: 14.5-36.5 h) in the low dose JHG group, 39.5 h (95% CI: 29.0-46.0 h) in the placebo control group. There was statistical difference in the median fever disappearance time between the low dose JHG group and the placebo control group (P = 0.011). Three days after treatment, the markedly effective rate of TCM symptoms in the low dose JHG group was 66.7%, higher than that of the placebo control group (38.3%), and its effective rate was superior to that of the high dose JHG group (P = 0.043). Five days after treatment, the recovery rate of the low dose JHG group (42.2%) was higher than that of the high dose JHG group (25.0%, P = 0.026) and that of the placebo control group (14.9%, P = 0.002). The markedly effective rate of the low dose JHG group (86.7%) was higher than that of the placebo control group (55.3%, P = 0.001). Similar effects were obtained in the low dose JHG group and the high dose JHG group, but slightly poor in partial indicators of the high dose JHG group. There was no statistical difference in adverse reaction among these three groups (P > 0.05).

CONCLUSIONSJHG was effective and safe in treating influenza patients of WHAFS. Routinely low dose was the optimal dosage of JHG.

Adult ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Influenza, Human ; diagnosis ; drug therapy ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Young Adult