Recombination plasmid pMM085 possessed both immunogens heat-labile enterotoxin(LT) and fimbriae antigen K88 of enterotoxigenic Escherichia coli (ETEC). Althouth vaccine strain MM-3 carrying pMM085 had good effect to protect piglets against diarrhea due to ETEC infections,it was not ideal live vaccine for pMM085 bringing chloramphenicol resistance gene (cat). To solve the problem,the host-plasmid balanced lethal system was introduced which including the replacement of cat gene by asd gene and transformation the new plasmid to the strain X6097 which asd gene was knocked out in its chromosome. Considering pMM085 was a big plasmid (23kb) and traditional genetic manipulations was not easy to carry on,?-Red recombination system was adopt in this work to realize the replacement of cat gene by asd gene. The results indicated that ?-Red recombination system was convenient and efficient to reconstruct big plasmid.

OBJECTIVETo study the color reappearance effect of metal-ceram restoration and foundry-ceram restoration using Crystaleye spectrophotometer.

METHODS58 metal-ceram restorations and 58 foundry-ceram restorations according to the result of the Crystaleye spectrophotometer were made respectively. The deltaE between restorations and natural teeth as referenced were analyzed. And satisfaction of dentists and patients were evaluated.

RESULTSThe deltaE between metal-ceram restorations and natural teeth was 7.13 +/- 0.74. The deltaE between foundry-ceram restorations and teeth was 1.47 +/- 0.84. There were statistical differences between the deltaE (P < 0.01). The satisnatural faction of dentists and patients to the foundry-ceram restorations were higher than to the metal-ceram restorations (P < 0.05).

CONCLUSIONCrystaleye spectrophotometer can provide accurate reference for foundry-ceram restoration, but for metal-ceram restoration it is not accurate.

Color ; Humans ; Metals ; Spectrophotometry ; Tooth

As one of the main water-soluble composites of Radix Salviae, salvianolic acid B is a phenolic acid ingredient of the Chinese drug, which is rich content in the herb and has strong pharmaceutical activity. It is used to treat cardiocerebral vascular diseases, antagonize hepatic/renal fibrosis, prevent cancer, and promote stem cell proliferation and differentiation. In the researches of its acting mechanisms, rather deepened studies have been carried out for its application on cardiocerebral vascular diseases, but that for others are rather fewer.
Benzofurans ; pharmacology ; therapeutic use ; Biomedical Research ; trends ; Disease ; Humans ; Medicine, Chinese Traditional ; trends ; Stem Cells ; drug effects ; Ventricular Remodeling ; drug effects

OBJECTIVETo discuss the options and clinical outcomes of surgical treatment of cervical ossification of the posterior longitudinal ligament (OPLL).

METHODSFrom January 2006 to December 2010, 63 patients of cervical OPLL treated surgically by same team were retrospectively analyzed. There were 49 males, 14 females, with a mean age of 56.2 years (from 38 to 76 years). The duration of symptoms ranged from 2 months to 20 years (mean 3.2 years). The ossified ligament was classified via sagittal and coronal images on CT scan. The cervical curvature was measured. Choice of surgical methods was determined according to the type of ossified ligament and cervical curvature. All complications occurred was recorded. The Japanese Orthopedic Association (JOA) scoring system was used to evaluate the neurological status. The severity of cervical pain was assessed with visual analogue scale (VAS) method. The results of preoperative, postoperative and follow-up were statistically compared with student t test.

RESULTSThe JOA scores of 35 patients performed with anterior approach improved from 9.8 ± 2.3 preoperatively to 11.7 ± 2.1 at final follow-up (t = 3.28, P < 0.05). The VAS sore was significantly decreased at final follow-up compared with preoperative (t = 3.15, P < 0.05). While the JOA scores improved from 10.4 ± 2.7 preoperatively to 12.5 ± 2.3 at final follow-up (t = 3.81, P < 0.05) in 12 patients with laminectomy and from 9.7 ± 2.5 to 11.6 ± 2.6 in 14 patients with laminoplasty (t = 3.56, P < 0.05). The VAS score either in laminectomy or in laminoplasty has not significantly difference between preoperative, 3 month postoperative and final follow-up (P > 0.05). Two patients underwent a combined anterior-posterior approach procedure. Complications in patients included 1 patient of postoperative neurologic deterioration and 2 cases of cerebrospinal fluid leakage in patients with anterior approach, and 2 cases of C(5) nerve palsy in patients with laminectomy.

CONCLUSIONSSurgical options of cervical OPLL should be determined by detailed study of type and range of ossified ligament, as well as the cervical curvature of patients. Good neurological function can be expected in case of appropriate choosing the method for treating the cervical OPLL.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Ossification of Posterior Longitudinal Ligament ; surgery ; Retrospective Studies ; Treatment Outcome

OBJECTIVESTo investigate the effects of newborn bull serum(NBS), vitamin C and vitamin E on cryopreservation of mouse seminiferous epithelial cells.

METHODSThe seminiferous epithelial cells from 7-day-old mice were cryopreserved in different freezing solutions. The cell recoveries were examined by Trypan blue exclusive staining after thawing. The freezing solutions composed of DMEM, 10% dimethylsulphoxide(DMSO), and 0, 5%, 10%, or 20% NBS, respectively, or composed of DMEM, 10% DMSO, 10% NBS, and 150 micrograms/ml vitamin C or 50 micrograms/ml vitamin E, respectively.

RESULTSThe cell recoveries in freezing solution containing 0, 5%, 10%, or 20% NBS were 83.4%, 84.7%, 85.7% and 83.6%, respectively. There were no significant differences between them. The cell recoveries in freezing solution containing vitamin C or vitamin E were 88.0% and 82.9%, respectively. There was no significant differences compared with that in freezing solution containing 10% DMSO and 10% NBS.

CONCLUSIONSNBS, vitamin C and vitamin E have no significant protecting effects on mouse seminiferous epithelial cells, and can not significantly improve the cell recoveries.

Animals ; Ascorbic Acid ; pharmacology ; Cattle ; Cryopreservation ; Epithelial Cells ; physiology ; Fetal Blood ; physiology ; Male ; Mice ; Seminiferous Epithelium ; cytology ; Vitamin K ; pharmacology

BACKGROUNDNon-cement femoral stems are recognized in clinical use, but there are still some problems. The aim of this research was to make non-cement femoral stems to be press-fit with the medullary cavity.

METHODSTwenty-four healthy adult mongrel dogs were randomly divided into experimental and control groups. In the right hip joint, an artificial femoral bone replacement surgery was conducted. For the experimental group, the replacement surgery of hydroxyapatite (HA)-coated femoral stems was done, while autogeneous morselized bone was implanted into the medullary cavity. For the control group, morselized bone was not implanted. At postoperative 1, 3, 6 months, a test for interfacial shear characteristics was conducted in the MTS810 Tester. The comparison between the two groups' bone-prostheses in shear strength for their interface from shearing destruction was made. A histological observation to check prosthesis-bone interface contact ratios and bone growth was carried out.

RESULTSFor the experimental group, shear strength was 0.317 MPa in 1 month, 1.447 MPa in 3 months, and 1.621 MPa in 6 months. For the control group, shear strength was 0.195 MPa in 1 month, 1.023 MPa in 3 months, and 1.483 MPa in 6 months. The difference was statistically significant. Stereomicroscope-based observation showed that the number of trabecular bones in the experimental group was larger than that of the control group, and bone growth of the former group was better than that of the latter group. Inverted microscopic observation showed that the binding degree between the prosthesis and trabecular bone of the experimental group was higher than that of the control group. Comparatively, the experimental group's trabecular bone had more stromal cells.

CONCLUSIONSThe morselized bones can effectively improve the biological bonding strength and bone-contact ratios in the short term for the HA-coated femoral stem and accelerate the bonding process. The use of morselized autogenous bones played a good role in bone in-growth of the femoral bone stem surface.

Animals ; Biomechanical Phenomena ; Coated Materials, Biocompatible ; chemistry ; Dogs ; Durapatite ; chemistry ; Female ; Femur ; pathology ; surgery ; Male ; Osseointegration ; Random Allocation ; Shear Strength

OBJECTIVETo explore the clinical effects of Stoppa approach and posterior percutaneous plate in treating type C pelvic fracture.

METHODSFrom June 2009 to July 2011,16 patients with type C pelvic fracture were treated with reconstruction plate fixation in Stoppa approach for anterior lesions combined with posterior percutaneous plate fixation for posterior lesions.There were 11 males and 5 females,with an average age of 38.8 years (ranged, 22 to 59 years). According to the Tile classification,10 cases belonged to C1,4 belonged to C2,2 belonged to C3. Tometta and Majeed score standards were used to evaluate clinical results.

RESULTSSixteen patients were followed up from 4 to 13 months with an average of 7.3 months. Operative time was from 80 to 140 min with an average of 100 min;blood loss volume was from 200 to 500 ml with an average of 280 ml; and the healing time of fracture was from 12 to 16 months with an average of 14 months. According to the Tometta score classification, 9 cases got excellent results, 6 good, 1 fair. According to the Majeed score classification, 9 cases obtained excellent results, 5 good, 2 fair.

CONCLUSIONReconstruction plate fixation in Stoppa approach for anterior lesions combined with posterior percutaneous plate fixation for posterior lesions is an ideal minimally invasive operation in treaing type C pelvic fracture. It can early exercise and has the advandages of small trauma, safe operaton,less complication, stable fixation.

Adult ; Bone Plates ; Female ; Fracture Fixation, Internal ; adverse effects ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; injuries ; surgery

OBJECTIVETo explore the synergistic protective effect of co-transplanted testicular cells expressing FasL and CsA on survival of islet allografts.

METHODSThe allogeneic islets and testicular cells were co-transplanted into the renal subcapsular space of the diabetic recipients with or without CsA after operation. Allografts survival period and the testicular cells or islets function were analyzed.

RESULTSThe mean survival period of control group was 4.6 +/- 1.1 days. When CsA was administered after transplantation, the mean survival period of islet allografts, (21.8 +/- 4.7) days, was significantly longer than that of control group (P < 0.01). When islets were co-transplanted together with 1 x 10(7) testicular cells (group A), a significant prolongation of graft survival was found (more than 57.5 +/- 4.0 days; P < 0.01 vs. control). But if 1 x 10(7) testicular cells expressing FasL were cultured with FasL-mAb for 30 minutes before co-transplantation (group B), the mean survival period of islet allografts (5.8 +/- 2.6 days), was similar to that in control group, but significantly shorter than that in group A (P < 0.01). When islets and 1 x 10(5) testicular cells were co-transplanted separately into the bilateral renal subcapsular space with CsA (group C), the survival of islet allografts was significantly prolonged in comparison with control group (more than 55.0 +/- 6.5 days; P < 0.01 vs. control), and similar to islets co-transplanted together with 1 x 10(7) testicular cells (group A). When islets were co-transplanted separately with 1 x 10(6) testicular cells without CsA (group D), the mean survival period (11.5 +/- 3.1 days) was shorter than that in group C, but prolonged in comparison to control group (P < 0.05).

CONCLUSIONThe co-transplanted testicular cells expressing FasL with administering CsA post-transplantation can jointly inhibit immune rejection of islet allografts by different mechanism and play a systemic and synergistic protective role to islet allografts.

Animals ; Cyclosporine ; therapeutic use ; Fas Ligand Protein ; Graft Survival ; Immunohistochemistry ; Insulin ; blood ; Islets of Langerhans ; pathology ; ultrastructure ; Islets of Langerhans Transplantation ; Male ; Membrane Glycoproteins ; analysis ; genetics ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Sertoli Cells ; metabolism ; transplantation ; Transplantation, Homologous

BACKGROUND & OBJECTIVE: Postoperative tissue adherence, scarring and radiotherapy often lead to extrinsic compression and stricture in the distal ureter of the patients who had history of pelvic malignancies. Our aim was to evaluate the efficacy and safety of endourologic techniques in treating this kind of ureteral obstruction. METHODS: From Jan. 1998 to Mar. 2007, 46 patients with obstruction in the distal ureter and had history of pelvic malignancies underwent endoscopic treatments at the Third Affiliated Hospital of Sun Yat-sen University for relief of the obstruction. Perioperative and follow-up data were analyzed. RESULTS: Of the 46 patients, 25 underwent laparoscopic ureterolysis and ureteroneocystostomy, 18 underwent placement of ureter stent under ureteroscope, 3 underwent percutaneous nephrostomy. No severe complication was recorded. The mean operating time was 82.5 min (range, 30-140 min). The mean blood loss was 45.5 ml (range, 5-180 ml). No blood transfusion was needed. The median follow-up time was 18.2 months (range, 3 months to 6.5 years). Three months after operation, B-ultrasonography and intravenous urography (IVU) showed that 39 (84.8%) patients had recovered normal renal function, the other 7 (15.2%) had hydronephrosis relief and renal function improvement. Nuclear renal scanning showed that the mean postoperative glomerular filtration rate (GFR) in the obstructive kidney was higher than the preoperative level (37.6 ml/min vs. 21.3 ml/min, P＜0.05). No stricture in the uretero-bladder anastomotic stoma was recorded. CONCLUSION: Endoscopic operation is an effective and feasible option for managing some selected kinds of distal ureteral obstruction caused by postoperative tissue adherence and radiotherapy in the patients with history of pelvic malignancies.

Objective To develop a simultaneous determination method of risperidone,paliperidone,olanzapine and quetiapine in human serum for therapeutic drug monitoring.Methods Clinical serum samples were prepared by protein precipitation and analytes were determined using HPLC-MS/MS.An Eclipse XDB C1s column (4.6 mm ×50 mm,1.8μm) was combined with mobile phase consisted of methanol,water and acetrile (37.5:12.5:50,plus 2.5 mmol · L-1 ammonium formate).The flow rate was 0.5 mL · min-1 The column temperature was 35 ℃ and the injection volume was 2 μL.MS detection was carried out using an electrospray ionization source operated in multiple reaction monitoring (MRM) mode.The ion transitions were m/z 427.3 → m/z 207.2(paliperidone),m/z 431.3--m/z 211.2 (paliperidone-D4),m/z 411→m/z 191 (risperidone),m/z 415.4→m/z195.2 (risperidone-D4),m/z 384.3→m/z 253.1 (quetiapine),m/z 392.3→m/z 258 (quetiapine-D8),m/z313→m/z 256 (olanzapine),m/z 316→m/z 256 (olanzapine-D3),respectively.Results The calibration curves of risperidone and paliperidone were y =0.67x +4.94 × 10-4 (R2 =0.999 4) and y =0.68x +0.14 × 10-2 (R2 =0.999 6),with the linear ranges of 1-100 ng · mL-1.The linear ranges were 2-200 ng · mL-1(calibration curve:y =1.33x-0.65 × 10-2,R2 =0.998 6) for olanzapine and 8-800 ng· mL-1 (calibration curve:y =1.80x-2.08 × 10-2,R2 =0.999 4) for quetiapine.The inter and intra-batch precisions of risperidone,paliperidone,olanzapine and quetiapine were all within 15 ％ in lower limit of quantitation (LLOQ),low,middle and high levels of quality control samples.The extraction efficiency was more than 90％ for four analytes.The concentration of analytes in quality control samples were less than ± 15 ％ bias compared with nominal concentration after storage at room temperature for 24 h,-70 ℃ for 34 d or after freeze-thaw for 3 cycles.Conclusion The validated method was simple,fast and sensitive for the simultaneous determination of risperidone,paliperidone,olanzapine and quetiapine in human serum.The method was applied to the therapeutic drug monitoring for patients administered with the above drugs.The stability of mixed quality control samples is reliable and can be used for therapeutic drug monitoring.