Objective To clone and express translocation intimin receptor(Tir)of enterohemorrhagic Escherichia coli(EHEC)O157:H7,and to analyze the effect of different routes on the induction of immunity to the recombinant protein.Methods The tir eucoding genes were amplified from EHEC O157:H7 strain guangzhou 246 genome,and genes were cloned into the vector pET-30a(+).The pET-30a(+)tir recombinant was transformed into E.coli BL21.and expression was induced bv IPTG.The expressed product was analyzed by SDS-PAGE and purified by Ni-IDA affinity chromatography.The immunized mice sera and fecal against the recombinant protein was detected.Resuits The length of the tir is 1677 bp,with the initiation codon ATG and the termination codon TAA.Double enzyme digestion and DNA sequencing confirmed that the recombinant expression plasmid pET-30a(+)-tir was constructed.The recombinant protein was expressed in Escherichia coli expression system,and was purified by Ni-IDA affinity chromatography.The mice were able to produce a high serum IgG antibody titer after both subeutaneous and intranasal immunizations.Meanwhile,the intranasal immunization induced serum and fecal IgA antibody titer was significantly higher than that of the subcutaneous immunization group.Conclusion Tir molecule is potential vaccine candidate for preventing EHEC disease.

Background Hyaluronic acid is a mucopolysaccharide existing in extracellular matrix and having good biocompatibility.Using chemical crosslinking method can improve the physical properties of the material,so cross-linked hyaluronic has potential clinical application value.Objective The present study was to evaluate the histocompatibility and biological security of cross-linked hyaluronic acid as ophthalmic implant material.Methods Cross-linked hyaluronic acid implant material was prepared according to the criteria of Biological Evaluation of Medical Devices (GB/T16886.5-2003).Eighteen 8-week-old male New Zealand white rabbits were randomized into the experimental group and the control group.Cross-linked hyaluronic acid material with 5.0 mm diameter was implanted into corneal stroma interlaminationally in the experimental group,and only corneal stromal interlaminational pocket was made without any implanting material in the control group.Biological response of cornea was assessed in vivo from 1 week through 3 months after operation by slit lamp microscope.The corneas were obtained 2 weeks,1 month and 3 months respectively for histopathological examination.Mouse embryonic fibroblasts were cultured in cross-linked hyaluronic acid film plate,medical silicone material culture plate and regular culture plate respectively for 24 hours,and the cell growth state and morphology were observed under the inverted microscope and scan electron microscope.MTT assay was used to test the relative growth rate of the cultured cells 48 hours after cultured using extracted liquid of hyaluronic acid implant material.Results Cross-linked hyaluronic acid implant material showed a well healing to the corneas of rabbits during the observation duration,without obvious inflammatory response and neovascularization.The arrangement of stromal fibers was uniform in order,and no infiltration of inflammatory cells was seen under the light microscope.The cells grew well after cultured with cross-linked hyaluronic acid film and regular medium for 24 hours,but in the silicone culture group,fewer of adherent cells and more floating cells were found.The relative growth rate of the cells was 87.50％ 48 hours after cultured with extracted liquid of hyaluronic acid implant material.Conclusions The cross-linked hyaluronic acid material has good histocompatibility and biological security in rabbit cornea tissue.

AIM: To investigate hemodynamic alterations of retrobulbar vessels in proliferative diabetic retinopathy ( PDR) patients with anterior segment neovascularization, before and 3mo after vitrectomy combined with panretinal photocoagulation and to explore the clinical significance.
●METHODS: Color Doppler flow imaging ( CDFl ) was used for measurement of blood flow velocities and resistive indexes ( Rl ) of the ophthalmic artery ( OA ) , short posterior ciliary arteries ( sPCA ) and central retinal artery ( CRA ) in 21 eyes of 21 PDR patients with anterior segment neovascularization. CDFl parameters were obtained before and 3mo after vitrectomy combined with panretinal photocoagulation ( PRP) .
● RESULTS: Peak systolic velocity ( PSV ) and end diastolic velocity ( EVD ) of CRA were significantly increased after surgeries, Rl were decreased significantly (P<0. 05). Parameters of sPCA and OA have no change after surgeries (P>0. 05).
●CONCLUSION: Vitrectomy combined with panretinal photocoagulation might increase the velocity of CRA, decrease Rl and improve ocular blood supply postoperatively. lt may delay or prevent the process of neovascular glaucoma.

Background: Dampness-heat syndrome is a major syndrome type in patients with a sharp deterioration of chronic renal failure (CRF). Qingshen Decoction, a compound traditional Chinese herbal medicine, could relieve the clinical symptoms of CRF patients, and was considered to have a certain reversal effect on rapid deterioration of renal function. Objective: To observe the changes of serum interleukin-8 (IL-8) and IL-18 levels in CRF patients with a sharp deterioration and dampness-heat syndrome, and to explore the curative efficacy of Qingshen Granule. Design, setting, participants and interventions: Sixty CRF patients with a sharp deterioration and dampness-heat syndrome from Department of Nephrology, the First Affiliated Hospital of Anhui College of Traditional Chinese Medicine, were randomly divided into treatment group (30 cases) and control group (30 cases), with another 20 healthy individuals as normal control. The patients in the treatment and control groups were all treated with Jiedu Xiezhuo II, a compound traditional Chinese herbal medicine, given as retention enema. Qingshen Granule was additionally administered to the patients in the treatment group with 1 dosage each time and 3 times a day. The treatment course was one month. Main outcome measures: The levels of serum IL-8 and IL-18 in the normal individuals and before and after treatment in the treatment and control groups were detected. Results: The total response rates of treatment group in clinical efficacy assessment and assessment of syndrome of traditional Chinese medicine (86.67% and 86.67%) were higher than those of the control group (56.67% and 60%), and there were significant differences between the two groups (P<0.05). The levels of serum IL-8 and IL-18 in CRF patients before treatment were obviously higher than those in the healthy individuals (P<0.01), and there were no differences in the levels of IL-8 and IL-18 before treatment between the treatment group and control group (P>0.05). After one-month treatment, the levels of serum IL-8 and IL-18 were markedly decreased in the two groups (P<0.01), and the levels of serum IL-8 and IL-18 in the treatment group were markedly lower than those in the control group (P<0.05). There were differences in decreased degrees of IL-8 and IL-18 levels between the two groups (P<0.05). Conclusion: CRF patients with a sharp deterioration and dampness-heat syndrome have high serum IL-8 and IL-18 levels. Qingshen Granule can reduce the levels of serum IL-8 and IL-18, and improve the renal function and ameliorate the clinical symptoms.

Adenoviridae ; genetics ; Calcium-Binding Proteins ; genetics ; metabolism ; Cells, Cultured ; Female ; Genetic Vectors ; Humans ; Microfilament Proteins ; genetics ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; Myometrium ; cytology ; metabolism ; RNA, Small Interfering ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

Objective To observe the anti-fibrosis therapeutic effect and mechanism of Qingshen granule for treatment of patients with chronic renal failure (CRF) accompanied by damp-heat syndrome.Methods Sixty-eight patients with CRF accompanied by damp-heat syndrome were randomly divided into a control group and a observation group, and the study was completed only in 61 patients, 31 in the control group and 30 in the observation group. Thirty subjects having taken physical health examination were assigned in a healthy control group. All the patients in both treatment groups were treated with conventional western medical therapy and traditional Chinese medicine (TCM) retention enema, and for patients in observation group, Qingshen granule was given additionally, 1 bag (10 g) thrice a day taken orally. The therapeutic course was 8 weeks. The clinical therapeutic effect, the levels of serum creatinine (SCr), the glomerular filtration rate (eGFR), serum interleukin-17 (IL-17), collagen type Ⅲ (Col-Ⅲ) and nuclear factor-κB p65 (NF-κB p65) in peripheral blood mononuclear cells (PBMC) were measured before and after treatment in the two treatment groups, and the above results were compared with those in healthy control group.Results Clinically, the total effective rates of the disease and of the TCM syndrome in observation group were significantly higher than those in the control group (86.67% vs. 58.06%, 83.33% vs. 45.16%, bothP < 0.01). In the observation group, the level of SCr was obviously lower, and the level of eGFR was markedly higher after treatment, and compared with the control group, the changes in above data after treatment in observation group were more significant [SCr (μmol/L): 250.62±164.97 vs. 393.72±183.64, eGFR (mL·min-1·1.73 m-2): 33.42±17.24 vs. 39.72±23.85, bothP < 0.05]. After treatment, the levels of serum IL-17, Col-Ⅲ and NF-κB p65 in PBMC were obviously lowered in both treatment groups compared with those before treatment, the therapeutic effect in observation group being superior to that in the control group [IL-17 (ng/L): 17.47±8.87 vs. 25.51±16.69, Col-Ⅲ (μg/L): 17.06±8.76 vs. 23.77±10.44, NF-κB p65 (μg/L): 0.58±0.34 vs. 0.83±0.30, allP < 0.05].Conclusion The Qingshen granule can ameliorate the clinical symptoms, improve renal function, decrease the levels of serum IL-17, Col-Ⅲ and NF-κB p65 in PBMC, intervene renal fibrosis in patients with CRF and damp-heat syndrome, ultimately delaying the progress of CRF.

Objective To detect the impact of reactive oxygen species ( ROS) on mitochondrial morphology and distri-bution during mitosis.Methods A viral vector in which the fluorescence gene was specifically under the control of mito-chondrial promoter was constructed and confirmed through DNA sequencing and Western blotting.After transfecting HeLa s3 cell with packaged virus, the HeLa s3-COX4tp-EGFP cell line stably expressing the mitochondrial fluorescence signal was obtained.With immunofluorescent staining, the impact of ROS on the morphology and distribution of mitochondria dur-ing mitosis was inspected.Result The cell line constantly expressing mitochondrial fluorescence signals was successfully constructed.Meanwhile,it was found that H2 O2 treatment could significantly change the morphology and distribution of mi-tochondria during mitosis by confocal microscopy.Conclusion Our study demonstrates that ROS can affect the morphology and distribution of mitochondria during mitosis.This research help study the relationship between the mitochondrial function and the regulation of mitosis in the future.

Objective To measure the reconstructed cranial CT images,and to clarify the safety range of unilateral nasal transsphenoidal approach for pituitary surgery.Methods 100 normal pituitary cranial CT images were randomly selected,and the three-dimensional reconstruction was performed by using the CT images of perpendicular and parallel to the edge of the two eyes as base line, and the distance and angle in unilateral nasal transsphenoidal approach for pituitary surgery from the sagittal plane in the middle of the nasal meatus and the plane through the tip of the nose and both ends of dorsum sellae were measured,respectively. The angles and distances were compared when grouped the data by gender and age. Results Angle A1 (the angle between the tip of the nose and the tuberculum sellae and saddle back root line in the sagittal plane)in the sagittal plane of the middle nasal meatus was (11.22±1.35)°,95% confidence interval was 8.92°-13.76°degrees;the distance D1(the distance on the line between tuberculum sellae and saddle back root, and the line was formed by the plate contained the angle A1 and sellar floor)was (16.71 ± 2.07)mm,95% confidence interval was 13.11-19.93 mm.Angle A2 (the angle between the tip of the nose and the saddle back ends)which was in the plane through the tip of the nose and both ends of dorsum sellae was (8.91±1.19)°,95% confidence interval was 7.12°-10.72°;the distance D2(the distance on the line between the saddle back ends,and the line was formed by the plate contained the angle A2 and sellar floor)was (14.23±2.09)mm,95% confidence interval was 10.81-17.92 mm. The four parameter data was normally distributed,and there was no significantly statistic difference between different gender and ages (P>0.05).Conclusion The angle of the movement for unilateral nasal transsphenoidal approach for pituitary surgery operation in the sagittal plane in the middle of the nasal meatus should be less than (11.22 ± 1.35)°,and the distance of the movement should be less than (16.71±2.07)mm. The angle of the movement in the plane through the tip of the nose and both ends of dorsum sellae should be less than (8.9 1 ± 1.1 9 )°, and the distance of the movement should be less than (14.23±2.09)mm.

AIMTo study the effect of XW630 on expression of pro-oncogene c-myc in the long bones of fetal mice in vitro for postulating the mechanism by which XW630 exerts its effect on bone.

METHODSThe fetuses of pregnant mice were removed on day 16 of gestation, the long bones of the forelimbs of female fetal mice were freed of muscle and soft tissue and cultured in a specific device for 48 h in BGJb medium treated with 1 x 10(-7), 1 x 10(-8) and 1 x 10(-9) mol.L-1 XW630 in the final medium. After cultured for 48 h, the long bones were harvested and immunohistochemical analysis was performed for determination of c-Myc protein expression in epiphyseal plates. The areas of positive cells in the resting zone, proliferative zone and hypertrophic zone in epiphyseal plate were determined under image analytic system.

RESULTSWhen the concentration of XW630 in the medium was 1 x 10(-9) mol.L-1, the area of c-Myc positive cells increased in the proliferative zone compared with 1 x 10(-9) mol.L-1 in the estrone group, significant increase was also observed in the resting zone compared with the control group. When the concentration of XW630 in medium was 1 x 10(-8) or 1 x 10(-7) mol.L-1, stronger expression than that in the control group and the estrone group at the same concentration was observed in each of the three zones.

CONCLUSIONThe estrogenic effect of XW630 on bone was stronger than that of estrone. XW630 may promote proliferation and differentiation of chondroncytes by promoting c-Myc protein expression in chondroncytes. Thus, endochondral bone formation was enhanced.

Animals ; Chondrocytes ; metabolism ; Culture Techniques ; Estrone ; pharmacology ; Female ; Fetus ; Mice ; Piperazines ; pharmacology ; Pregnancy ; Proto-Oncogene Proteins c-myc ; metabolism ; Tetracyclines ; pharmacology ; Ulna ; drug effects ; metabolism ; Up-Regulation ; drug effects

OBJECTIVETo explore the operative methods and effects of treatment of old acetabular fractures.

METHODSFrom October 2001 to October 2007, 26 patients with old acetabular fractures were treated with operation including 21 males and 5 females with an average age of 34 years ranging from 18 to 65 years. On the basis of the three-dimensional computed tomography, all cases were diagnosed and classified according to Letourne-Judet classification, 9 cases were posterior wall fracture, 3 cases were lateropulsition fracture, 7 cases were lateropulsition and posterior wall fracture, 2 cases were posterior column and posterior wall fracture, 2 were T-shape fracture, 3 were dual column fracture. These patients were treated through the anterior,posterior, combined anterior-posterior approaches. The time from injured to operation was 33 to 141 days (averaged 36.4 days). All the fractures were fixed with screws and AO reconstruction plates.

RESULTSAll patients were followed up for 6 to 96 months, with an average time of 32.4 months. Evaluated according to Matta criteria, the results of scores was (5.04 +/- 1.04) on pain, (5.23 +/- 0.76) on range of motion, (4.92 +/- 1.16) on walking,and tatal (5.06 +/- 0.99) on average; The functional results of hip joints were excellent in 6 cases, good in 10 cases, fair in 6 cases, and poor in 4 cases. Sciatic nerve injury was found in 2 patients,lateral femoral cutaneus nerve injury in 3 patients, necrosis of femoral head in 1 patient,infection in 1 patient, and ectopic bone formation in 6 patients.

CONCLUSIONGood clinical results can be obtained by careful selection of operative indications of old acetabular fractures in combination with proper operative approach and correct reduction and fixation.

Acetabulum ; injuries ; surgery ; Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; Time Factors ; Young Adult