The anisotropic diffusion equations-Perona &Malik(P&M)equations are resorted to to decrease the Rician noise introduced into the diffusion weighted(DW)images.To evaluate the efficiency of the P&M equations(with different functions)in accounting for the Rician noise,many experiments are designed.The results acquired from both Monte Carlo simulation experiments and real data based experiments proves the better performance of the reciprocal function based P&M equation in denoising the DTI images.

Objective To detect Chlamydia trachomatis phage Vp1 gene in clinical swab specimens and anti-Vp1 antibodies in serum specimens.MethodsCervical and urethral swab as well as serum specimens were collected from attendees to the sexually transmitted disease(STD) clinic in the Tianjin Institute of STD,during March 2008 to March 2011.PCR was conducted to detect chlamydial phage Vp1 gene in swab samples,enzyme linked immunosorbent assay(ELISA) and Western blot to detect anti-Vp1 antibody in sera.The swab specimens positive for Vp1 gene were subjected to cell culture followed by the detection of Vp1 protein with an immunofluorescence-based method.ResultsTotally,36 out of 1542 swab specimens turned out to be positive for Vp1 gene,and 23 out of 453 serum specimens for anti-Vp1 antibody.No positive results were obtained in the Vp1 gene-positive swab specimens by cell culture and immunofluorescence-based assay.ConclusionThe Vp1 gene of Chlamydial trachomatis phage and anti-Vp1 antibody are successfully detected from clinical swab and serum specimens respectively.

BACKGROUND: There are few reports about vascularization in the repairing of bone defect by heterogeneous deproteinized bone.OBJECTIVE: To verify the vascularization characteristics of heterogeneous deproteinized bone, tissue engineering scaffold material, in the repairing of large segmental bone defect.DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed between March 2005 and February 2007 at the Third Military Medical University, Chongqing, China.MATERIALS: Twenty-four 10 to 12 months old goats, weighing (22.5±2.5)kg, were obtained from the Animal Center of the Third Military Medical University, Chongqing, China. Segmental bone defects of 20 percent right tibia middle and inferior diaphysis of the 24 goats were made.METHODS: The 24 goats were divided into test group (n=16) and control group (n=8) randomly. Goats in test group were implanted with deproteinized bone+autologous MSCs+recombinant human bone morphogenetic protein-2 (rhBMP-2), goats in control group were implanted with autograft bone, and all fixed with half-ring sulcated external fixator. Every 4 weeks, 3 goats were killed after ink perfusing through femoral artery. A thick slice of new bone tissue was made to observe the vascularization.MAIN OUTCOME MEASURES: Vascularization of new bone observed through gross anatomy and imaging; vascular network of new bone observed through thick section, blood vessel amount and area measured by Image-proplus really image analysis software.RESULTS: No goat was infected or dead. Animal soft tissue was dyed black, blood vessels'size, ditribution and network structure were observed in subcutaneous tissue, fascia and periosteum. At 4 weeks postoperation implant margin became crude in the defect area; at 8 weeks postoperation transparent bone absorbing area of different size and irregular shape appeared; after 12 weeks postoperation high-dense calcification shadow appeared at the ends of defect bone and new bone connected with the ends completely. On 4 to 24 weeks postoperation, the blood vessel amount became large, alignment became regular, and their size and distribution became uniform. It showed no significant difference in blood vessel amount and area between 2 groups (P>0.05).CONCLUSION: Composite of heterogeneous deproteinized bone+autologous MSCs+rhBMP-2 has the same vascularization degree. of autogeneous bone graft in repairing of large segmental tibia defect.

Objective To observe the changes of CD4+CD25+ regulatory T cells, Foxp3 mRNA and soluble interlukin 2 receptor (sIL-2R) in the peripheral blood of kidney transplantation recipients and to evaluate their effect on the diagnosis of acute rejection. Methods Forty-two renal transplant recipients and 30 healthy controls were enrolled in this study. CD4+CD25+ regulatory T cells proportion, Foxp3 mRNA and sIL-2R of pre-transplantation and those of day 7,14, 28, 56 of post-transplantation were measured by flow cytometer, fluorescent quantization PCR and enzyme-linked immunosorbent assay (ELISA), respectively. Biochemistry appliance was used to detect serum creatinine. The diagnosis of acute rejection in transplanted kidney was based on the clinical symptoms, the laboratory examinations, Doppler ultrasound and biopsy. Results (1)At day 7, 14, 28, 56 of post-transplantation, CD4+CD25+ regulatory T ceils proportion, Foxp3 mRNA level in acute rejection group were significantly decreased compared with those in non-acute rejection group. (2) There were significant differences of peripheral blood CD4+CD25+ regulatory Tcells[(9.22±3.53)% vs (6.09±1.99)%, P<0.01], Foxp3 mRNA[(0.82±0.36)×10-3 vs (0.50±0.28)×10-3, P<0.01] and sIL-2R levels [(856.30±108,24) U/ml vs (247.35±11.24) U/ml, P<0.01]between patients of pre-transplantation and healthy control group. (3)Plasma CD4+CD25+ regulatory T cells [(16.53±4.14)%] and the expression of Foxp3 mRNA [(4.97±1.94)×10-3] was significantly increased, but sIL-2R level [(463.72±31.23) U/ml] was significantly decreased as the transplanted renal function was restored (all P<0.01). (4) Plasma CD4+CD25+regulatory T cells [(12.18~2.86)%] and the expression of Foxp3 mRNA [(3.15±1.22)×10-3] was significantly decreased (P<0.01), and sIL-2R level [(748.36±115.41) U/ml] was significantly increased (P<0.01) when acute rejection occurred. The above changes had an earlier onset than the change of Scr. (5)The percentage of CD4+CD25+ regulatory T cells was positively correlated with the Foxp3 mRNA level (P<0.01), but was not correlated with sIL-2R level in all the patients. Conclusion The measurement of these markers in peripheral blood may be an important guideline to the diagnosis and prognosis of acute rejection in renal transplant recipients.

BACKGROUND:Studies have demonstrated that tetrandrine has protection on acute spinal cord injury,but the specific mechanism remains poorly understood.OBJECTIVE:To study the protection of tetrandrine on rat acute spinal cord injury and to study its mechanism from apoptosis pathway.METHODS:A total of 100 rats were randomly divided into 4 groups.All rats were prepared for spinal cord injury models using modified Allen method except that in the sham-surgery group.Methylprednisolone and tetrandrine was injected into rats in the methylprednisolone and tetrandrine groups by tail intravenous injection prior to and at 24,48 hours after model preparation.The same volume of physiological saline was injected in the sham-surgery and model groups.Basso-BeatUe-Bresnahan(BBB score)was recorded at 8 hours,1,3,7 and 14 days after model preparation.The morphological changes of spinal cord injury sites were observed by hematoxylin-eosin staining and the expressions of bcl-2 and bax were determined by immunohistochemistry.RESULTS AND CONCLUSION:The BBB score of methylpradnisolone and tetrandrine groups were significantly higher than that model group at 7 and 14 days(P<0.05),but there were no significant difference between the methylprednisolone group and tetrandrine group(P>0.05).Hematoxylin-eosin staining showed that the spinal cord injured severely at 3-7 days,the injury degree in the methylpradnisolone group and tetrandrine group was slighter than that of the model group,with smaller bax expression and greater bcl-2 expression(P<0.01).The findings demonstrated that,tetrandrine is able to protect neurons from apoptosis and promote the nerve function recovery by inhibiting the expression of Bax and promoting the expression of Bcl-2.Its effect is not inferior to methylprednisolone.

ObjectiveTo explore the clinical outcome of one stage posteroanterior decompression and bone implant in the treatment of severe lower cervical spinal bony canal stenosis. Methods The study involved 29 patients with severe lower cervical spinal bony canal stenosis treated with one stage posteroanterior decompression and bone implant from April 2006 to March 2009. There were 11 patients with old fractures, seven with posterior longitudinal ligament ossification and 11 with cervical disc calcification. The course of disease ranged from 2 months to 3.2 years, average 1.4 years. The nerve function was rated as grade B in two patients, grade C in 19 and grade D in eight according to Frankel scale. The average Japanese Orthopaedic Association (JOA) score was 9.8. ResultsAll patients were followed up for 7-28 months (average 15.2 months), which showed bony fusion five months after operation, with fusion rate of 100％. The Frankel grade was increased for average 1.2 grades and the nervous symptoms alleviated remarkably. Mean postoperative JOA score was 13.8 and increased for mean 4.0, with mean amehoration rate of 55.6％. ConclusionsOne stage posteroanterior decompression and bone implant is a safe and effective method for treatment of lower cervical spinal bony canal stenosis, when the intraoperative electrophysiological monitoring can assure the operative safety.

Objective To clone, express, and identify chlamydial GPIC capsid Vpl gene and pro- tein. Methods The complete sequence of Vp1 gene from?CPG1 phage was amplified. The amplicor was cut by restriction endonuclease, linked to plasmid vector, and transformed into E. coli.The expressed pro- tein of recombinant Vp1 was purified and identified. Results The recombinant 1 661 bp gene was se- quenced and proved to be?CPG1 Vp1 by searching Genebank. A 62 kDa capsid protein was expressed and confirmed by SDS-PAGE and Western blot. Conclusion The recombinant Vp1 seems to be a highly con- served and specific marker for chlamydial phage.

OBJECTIVETo generate an HPV16 prophylactic vaccine candidate for cervical cancer.

METHODSHPV16 major capsid protein L1 gene and minor capsid protein L2 gene were amplified using PCR. These genes were mutated by PCR site-directed mutagenesis for removal of sequence motifs (TTTTTNT) which would cause transcription termination when expressed from a vaccinia virus early promoter, then inserted into a vaccinia virus expression vector. A strain replication-deficient recombinant vaccinia virus containing the mutant sequences was obtained through a homologous recombination and identified.

RESULTSThe nucleotide sequence remained the correct amino acid sequence of the L1 and L2 proteins after mutated. Full-length L1 and L2 proteins were generated in cells infected with the recombinant virus. The virus strain propagated at very low titer or could not reproduce in some kinds of cell derived from different human tissues.

CONCLUSIONSThe authors have generated a strain replication-deficient recombinant vaccinia virus expressing HPV16 L1 plus L2 proteins as an HPV16 prophylactic vaccine candidate for cervical cancer.

Capsid ; Capsid Proteins ; genetics ; Cell Line ; Cloning, Molecular ; Female ; Gene Expression ; Genetic Vectors ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; physiology ; Papillomavirus Infections ; prevention & control ; Transfection ; Tumor Virus Infections ; prevention & control ; Uterine Cervical Neoplasms ; virology ; Vaccinia virus ; genetics ; physiology ; Virus Replication

OBJECTIVETo generate a candidate HPV16 vaccine for experimental and therapeutical use for cervical cancer.

METHODSThe mutants of HPV16 early E6 and E7 genes were inserted into a vaccinia virus expression vector. A strain of recombinant vaccinia virus was constructed through homologous recombination.

RESULTSShowed that the mutant E6 and E7 genes were located at TK gene region of vaccinia virus Tiantan strain in a head to head orientation under the control of early/late promoters, H6 and 7.5K respectively. Studies in mice indicated that VmE6E7 could elicit specific antibodies against E6 and E7, and retarded or prevented tumor development in a proportion of C57 BL/6 mice challenged by syngeneic HPV16E6 and E7 transformed tumor cells.

CONCLUSIONSThe success in constructing VmE6E7 provides a basis for the further development of HPV16 therapeutic vaccine.

Animals ; Female ; Genes, Viral ; genetics ; Genetic Vectors ; genetics ; immunology ; Mice ; Mice, Inbred C57BL ; Mutation ; Neoplasms, Experimental ; prevention & control ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Recombination, Genetic ; Repressor Proteins ; Transfection ; Vaccinia virus ; genetics ; immunology

Objective To investigate the safety and efficacy of LVIS stent combined with coil embolization of ruptured wide-necked intracranial aneurysms during the acute phase.Methods From May 2014 to August 2017,the clinical and imaging data of 56 patients with ruptured wide-necked intracranial aneurysm treated with LVIS stents for acute phase assisted embolization at the Department of Neurosurgery,the Affiliated Hospital of Southwest Medical University were analyzed retrospectively.All patients were treated with LVIS stent combined with coil embolization.Immediate postoperative angiography,six months after procedure,and follow-up imaging were evaluated by Raymond grade (RS grade).The clinical follow-up results were evaluated by the modified Rankin Scale (mRS) score.Results LVIS stent combined with coil embolization was performed in 56 patients with 60 aneurysms in this group.The success rate of stent release was 100％.Immediate angiography after procedure showed that the complete embolization rate of aneurysms was 80.0％ (48/60),the near complete embolization rate was 13.3 ％ (8/60),and the incomplete embolization rate was 6.7％ (4/60).Postoperative follow-up angiography at 6 monthrevealed that the complete embolization rate of aneurysms was 87.8％ (36/41),nearly complete embolization rate was 7.3％ (3/41),incomplete embolization rate was 4.9％ (2/41).Postoperative follow-up angiography at 12 months revealed that the complete embolization rate of aneurysms was 83.0％ (39/47),and near complete embolization rate was 12.8％ (6/47),and incomplete embolization rate was 4.3％ (2/47).Of the 56 patients,49 were followed up clinically and 7 were lost to follow up.The average follow-up time was 13 ± 4 months.The clinical follow-up showed that the good prognosis (mRS score 0-2) rate was 87.8％ (43/49).Intraoperative complications occurred in 7 cases,5 were intraoperative parent artery thrombosis and 2 were intraoperative aneurysm rupture.Conclusions LVIS stent combined with coil embolization of ruptured wide-necked intracranial aneurysms during the acute phase has good efficacy and safety.Its long-term efficacy remains to be confirmed by long-term follow-up.