0.05).Conclusions RFA treatment of PHC patients does not lead to hematogenous tumor dissemination.

Transforming growth factor-?_1 (TGF-?_) mRNA or protein expression in renal cortex of diabetic rats was assessed by real-time quantitative RT-PCR with SYBR Green,immunohistochemistry or Western blot.After melatonin treatment,the expressions of TGF-?_1 mRNA and protein were decreased,suggesting that beneficial effect of melatonin may result from its antioxidative property and inhibiting TGF-?_1 expressions.

Objective To develop a nylon membrane chip for rapid and systematic detection of the diabetes-associated 45 mutant loci in mitochondrial DNA(mtDNA).Methods The mutant-and wild-type probes were designed for detection of 45 mutant loci in mtDNA with Primer Premier 5.0 and NCBI BLAST softwares and the 90 probes with 8 poly T were immobilized on the Hybond N~+ nylon membranes which were treated with 5?SSC Buffer by UV-crosslinking;Then asymmetric PCR was employed to obtain the target single strand DNA(ssDNA).The PCR products were labeled with biotin after purification.NBT/BCIP was used as substrate that yields a very intense purple signal followed by AP-avidin,and the signals were observed in 24 samples with known sequences to evaluate the chips,each sample was repeatedly measured three times.Results The specific target fragments of 45 loci can be amplified under the same condition with nine sets of primers.The annealing temperatures of the wild-type [(59.01?1.42)℃] and mutant-type [(59.34?1.29)℃ ] probes are so close(t=1.046,P =0.301)that hybridization can be performed at the same temperature.The spots on the membrane chip are distinct,regular and well-distributed.The results of positive-and negative-control are perfect.The signals of negative probes and the background are similar.The results of chip were nearly concordant with that of DNA sequences(?~2=113.132,Kappa value =0.888,P = 0.000)except for T16189C mutant.Conclusions We have successfully developed a nylon membrane chip for rapid and systematic detection of the diabetes-associated 44 mutant loci in mtDNA.It could be used for screening for diabetic patients and high-risk people.

OBJECTIVETo analyze the influence of included angle between the anterior aspects of S2 and S vertebral bodies on pelvic inlet imaging in the pelvic midline sagittal plane.

METHODSTotally 58 axial pelvic CT scans were chosen as study objects including 43 males and 15 females,with an average age of 40.7 years old (ranged,18 to 68 years old). The angles between the anterior aspects of S2 and S1, vertebral bodies and the horizontal plane on midline sagittal CT reconstruction were measured to simulate the optimal S2 and S1 inlet angles. The included angle between the anterior aspects of S2 and S1 vertebral bodies was calculated by subtrocting the S1,inlet angle from the S2 inlet angle defined as a base number. Then, the impact of the calculated included angles on the pelvic inlet imaging was analyzed. Results:The S2 inlet angles averaged (30.5±6.5) degrees; the S inlet angles averaged (25.7±5.9) degrees. The difference between them was significant (t=3.35, P=0.001). Ten patients had zero angle between the anterior aspects of S2 and S1 vertebral bodies; 14 patients had negative angle, averaged-(8.9±8.1) degrees; 34 patients had positive angle,averaged (11.8+6.4) degrees.

CONCLUSIONThe difference of included angle between the anterior aspects of S2 and S1 vertebral bodies leads to the difference between S1 inlet view and S2 inlet view in most cases, complicating the pelvic inlet imaging,and affecting the reliability of the application of pelvic inlet view. Utilizing the angles measured on the preoperative midlihe sagittal CT reconstruction to obatin the patient-customized S1 and S2 inlet views could accurately guide the S1 and S2 iliosacral screw insertion.

Adolescent ; Adult ; Aged ; Animals ; Bone Screws ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Image Processing, Computer-Assisted ; Male ; Middle Aged ; Pelvis ; anatomy & histology ; injuries ; Spine ; anatomy & histology ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVESTo introduce a classification system of upper sacral segment and its significance based on the continuous pelvic axial computed tomography scan.

METHODSThe whole pelvis 2.0 mm thick axial scan images of 127 cases were observed, the sacroiliac screw channel of S1 were measured, according to the size of the transverse screw channel the upper sacral segment were classified. Such as transverse screw channel existed and in at least 4 layer scan images its width was > 7.3 mm, it was defined as sacral segment of the normal type. Such as transverse screw channel existed and its maximum width was 7.3 mm or less on scanning level, it was defined as a transitional. Such as transverse channel did not exist, or its width on all scanning level was 0 mm or less, it was defined as dysplastic. Various cases,percentage, and the average of the transverse screw channel were calculated.

RESULTSThere were 58 normal (45.7%),42 transitional (33.1%), and 27 dysplastic (21.2%) upper sacral segments with an averaged width of the tansverse screw channel of 13.9 mm, 5.2 mm, and 0.9 mm, respectively. Each specimen could be defined as one of the three types of upper sacral segment without exceptions.

CONCLUSIONIt is possible to insert a transverse iliosacral screw into a normal upper sacral segment when indicated because of the capacious transverse screw channel. The transverse iliosacral screw placement into the transitional and dysplastic upper sacral segments was contraindicated because of the limited or none transverse screw channel. The transitional upper sacral segment was superior to the dysplastic segment due to its starting point location restriction on the true lateral sacral view.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Density ; Bone Screws ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Pelvic Bones ; diagnostic imaging ; surgery ; Sacrum ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVESTo study whether Chinese Medicine Yiqihuoxuetang(YQHXT) could inhibit antisperm antibodies in infertile men, and to explore the therapeutical mechanism of YQHXT.

METHODSThirty infertile men with antisperm antibodies took YQHXT continuously for 60 days. Indirect immuno-fluorescence technique (IFT) was used to detect the levels of CD3, CD4, CD8 and CD4/CD8 ratio before and after treatment.

RESULTSCD4 value and CD4/CD8 ratio after treatment were significantly lower than before treatment (P < 0.05); CD8 value became significantly higher(P < 0.05).

CONCLUSIONSThe results indicated that YQHXT could inhibit antisperm antibodies by keeping the balance of T-lymphocyte subpopulation in immunoinfertile men.

Adult ; Autoantibodies ; immunology ; CD4-CD8 Ratio ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Infertility, Male ; immunology ; pathology ; Male ; T-Lymphocyte Subsets ; drug effects

OBJECTIVETo develop a 96-microwell plate DNA diagnostic chip for simultaneous detection of 9 major foodborne bacteria.

METHODSType-specific PCR primers labeled with biotin and oligonucleotide probes were designed according to the conservative genes of 9 major foodborne bacteria Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7 (Stx1 and Stx2), Shigella spp., Listeria monocytogenes, Bacillus cereus, Yersinia enterocolitica, Vibrio cholerae and Vibrio parahaemolyticus. A one-tube multiplex PCR system for simultaneous amplification of these bacteria was established, and the DNA probes were spotted and immobilized in the wells of the plate in 5x5 array format. Stable hybridization system between PCR products and oligonucleotide probes in the microwell was established after condition optimization. Alkaline phosphatase-conjugated streptavidin and NBT/BCIP were used to detect the hybridized PCR products.

RESULTSTwenty standard bacteria strains were used to validate the 96 microwell plate DNA diagnostic chip and highly specific and stable experiment results were obtained. Using this chip assay, the causal pathogen Staphylococcus aureus was identified within 12 h after the sampling from an incident of food poisoning, and the result was consistent with that obtained using conventional bacterial culture and biochemical identification.

CONCLUSIONThe novel 96 microwell plate DNA diagnostic chip allows rapid, accurate, automated and high-throughput bacterial detection and is especially valuable for quick response to such public health emergencies as food poisoning.

Bacteria ; classification ; genetics ; isolation & purification ; DNA, Bacterial ; analysis ; Food Contamination ; analysis ; Food Microbiology ; methods ; Foodborne Diseases ; microbiology ; Humans ; Oligonucleotide Array Sequence Analysis ; methods

OBJECTIVETo initially evaluate the application of artificial vertebra of n-HA/PA66 in anterior reconstruction of lower cervical spine fracture and dislocation.

METHODSIn this study, 84 patients with lower cervical spine fracture and dislocation received anterior cervical discectomy, spinal canal decompression or subtotal corpectomy, spinal canal decompression and reconstruction by n-HA/PA66 composite artificial vertebral body combined with plate instrumentation. Neurological function was followed up by improvement rate of Frankel and situations of the supporting body was observed by X ray and 3D-CT in 3, 12, 24 months postoperatively. The intervertebral height, physical arc (reflected by Cobb angle) and the locations and fusion rate of the supporting body were assessed in order to evaluate the stability of the cervical spine and alignment improvements.

RESULTSAll the patients underwent operation successfully and were followed up for 6 to 24 months with an average of 12 months. The preoperative symptoms were improved to varying degrees. Imaging studies showed that in all cases graft fusion were achieved, and cervical alignments, intervertebral height, cervical spine stability and the locations of the artificial vertebral body were well maintained. No displacement and subsidence of the artificial vertebral body occurred. Postoperative immediate intervertebral height (2.4 ± 0.2) cm, preoperative intervertebral height (1.9 ± 0.1) cm, comparisons of the two groups was statistically significant (q = 2.48, P < 0.001). The immediate, 3 month, 1 year, 2 year period follow-up group intervertebral height was not statistically significant (P > 0.05). Preoperative Cobb angle was 9.8° ± 1.2°, postoperative immediate Cobb angle was 16.6° ± 1.2°, comparisons of the two groups was statistically significant (q = 14.25, P < 0.001). The immediate, 3 month, 1 year, 2 year period follow-up group Cobb angle was not statistically significant (P > 0.05).

CONCLUSIONSn-HA/PA66 artificial vertebral body can provide early cervical spine support and stability and effectively maintain the biological alignment and cervical intervertebral height. It has high rate of graft fusion and is convenient to observe by X-ray. Therefore, n-HA/PA66 can be taken as an ideal graft for anterior lower cervical spine fracture and dislocation operation, but further follow-up study is still required to evaluate the long-term effects.

Adolescent ; Adult ; Aged ; Bone Substitutes ; Cervical Vertebrae ; injuries ; surgery ; Decompression, Surgical ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Humans ; Hydroxyapatites ; Joint Dislocations ; complications ; surgery ; Male ; Middle Aged ; Nanostructures ; Nylons ; Spinal Fractures ; complications ; surgery ; Spinal Fusion ; instrumentation ; Young Adult

OBJECTIVETo developed a multiplex RT-PCR assay for simultaneous screening of type A, B and novel A (H1N1) influenza viruses.

METHODSTwo pairs of universal primers in were designed for amplifying the M gene and NS gene of type A and B influenza viruses, respectively. A pair of specific primers of HA gene was designed to detect novel A (H1N1) influenza virus. A one-step method was used to establish the multiplex RT-PCR system. A blinded experiment was carried out to validate the accuracy of this assay in comparison with the results of real-time fluorescence RT-PCR. The clinical practicability and efficacy of this assay was also evaluated.

RESULTSThe RT-PCR products were analyzed using agarose gel electrophoresis, which yielded distinct bands of the target fragments without non-specific reactions, suggesting the high efficiency and specificity of the multiplex RT-PCR. Blinded study of 50 samples demonstrated a concordance rate of 100%.

CONCLUSIONThis multiplex RT-PCR assay allows one-step simultaneous detection of type A, B and novel A (H1N1) influenza viruses rapidly and accurately, and provides a valuable low-cost screening technique for influenza epidemic monitoring and early diagnosis.

Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza B virus ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Time Factors ; Viral Matrix Proteins ; genetics ; Viral Nonstructural Proteins ; genetics

OBJECTIVETo determine the self-compatibility of Platycodon grandiflorum and the location of microspore germination on stigma.

METHODThe microspore germination of self-pollination, self-plant-pollination and cross-pollination and the pollination microspore germination in and outside of stigma were observed with fluorescence microscopy.

RESULTMost pollens of self-pollination, self-plant-pollination and cross-pollination can germinate on the stigma, and after 24 hours, pollen tube entered into the ovary successfully. Pollinated on the outer-surface of stigma, microspores could not germinate, but on the inner-surface of the stigma when it dehisced most microspores can germinate.

CONCLUSIONThe compatibility of self-plant-pollination of Platycodon grandiflorum is high. The microspore germination loci is on the inner-surface of the dehisced stigma.

Flowers ; growth & development ; Microscopy, Fluorescence ; methods ; Plants, Medicinal ; growth & development ; Platycodon ; growth & development ; Pollen Tube ; growth & development ; Pollination ; physiology