Traditional Chinese Medicine(TCM) makes great contributions to the prosperous growth and people's health.But understanding deviation and imperfect evaluation system of TCM affect the healthy development of TCM.Clinic practice is the motive power of TCM,and curative effect is the key of TCM researches,and the scientific evaluation system is the safeguard for a healthy development of TCM.So we should focus on clinical researches of stubborn diseases and emergency cases to satisfy social demand and upgrade the position of TCM in the medical system.At the same time,functional disease must be explored to show the advantage of TCM.Our mission is to establish a scientific objective evaluation system to accurately understand TCM and take it as the turning point to give an impetus to theoretical breakthrough of the basic studies to promote an overall and healthy development of TCM.

OBJECTIVETo explore methods of treating middle and distal tibia nonunion with the treatment of advanced bone graft combined with locking compression plate.

METHODSFrom January 2011 to December 2012, 12 patients with middle and distal tibia nonunion were treated with advanced bone graft combined with locking compression plate. Among patients, there were 8 males and 4 females aged from 20 to 69 with an average of 47 years old. The time from first injuries to bone nonunion was from 9 months to 5 years, avergaed 19 months. Four cases were treated with external fixation, 6 cases were treated with plate fixation, 2 cases of 12 patients occurred broken of plate and nail. Eleven patients were non-infective bone nonunion and 1 patient was infective bone nonunion. Preoperative X-ray and CT showed all patients had sequestration and formation of ossified bone with different degrees. Operative time, blood loss, wound healing were observed, fracture healing time was evaluated by postoperative X-ray. Johner-Wruhs scoring standards was used to evaluate ankle joint function after operation at 10 months.

RESULTSOperative time ranged from 90 to 185 min with an average of (125.00±20.15) min; blood loss ranged from 225 to 750 ml with an average of (415.00±120.00) ml. All patients were followed up from 10 months to 2.5 years with an average of 1.5 years. Postoperative X-ray showed bone union was formed around fracture after operation at 4 months in all patients, 3 cases obtained bone healing within 6 months after operation, 9 cases obtained from 8 to 12 months. No infection, injury of nerve and vessles, and broken of plate and nail were ocurred. According to Johner-Wruhs scoring at 10 months after operation, 10 cases obtained excellent results, 1 good and 1 moderate.

CONCLUSIONAdvanced bone graft combined with locking compression plate, which can build fracture multi-point supporting based on full compression of bone nonunion to get effective fixation, is an effective method in treating middle and distal tibia nonunion.

Adult ; Aged ; Bone Plates ; Bone Transplantation ; Female ; Fracture Healing ; Fractures, Ununited ; surgery ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery

In this paper, Gaussian pixelate Chinese character processing software is designed, and HMD is used to realize the recognition experiment for pixelate Chinese characters based on simulated prosthetic vision. The structure of recognition system, software design and the experiment for determining Gaussian width (sigma) are presented. It is shown that when sigma is 0.235, the recognition program is the best.
Equipment Design ; Image Processing, Computer-Assisted ; Language ; Prostheses and Implants ; Software Design ; Visual Perception

In this article, the implementation of eye movement tracking system includes three procedures: hardware acquisition, data extraction and overall analysis. The system is based on Camshift algorithm with an eye tracking module added, developed on VC++ 6.0. The system can track the eye movement effectively in simulated phosphene evaluation experiment based on prosthetic vision.
Algorithms ; Analysis of Variance ; Eye Movements ; physiology ; Prosthesis Design

OBJECTIVETo explore morphological character and clinical significance of superior-posterior acetabular wall by anatomically measuring and quantitatively analyzing thickness of posterior acetabular wall, then provide a theoretical reference for clinical treatment of acetabular fracture.

METHODSFifteen adult formalin-preserved cadaveric pelvises (8 males and 7 females) were used for this investigation. Excess soft tissue was removed and the whole acetabular posterior walls were marked with "angle" sector method and the thickness was measured with caliper in different levels of the different split points. The measurement results were validated and analyzed statistically.

RESULTSAt 5 mm away from acetabular rim, the average thickness of superior-posterior acetablar wall fluctuated between (6.47±0.61) mm and (7.43±0.71) mm; the average thickness of inferior-posterior acetabuluar wall fluctuated between (5.62±0.51) mm and (6.33±0.61) mm; the average thickness of acetabular roof fluctuated between (7.71±0.74) mm and (8.27±0.99) mm. There was no statistical difference between average thickness of superior-posterior wall of acetabulum and inferior-posterior wall of acetabulum (P>0.05), but the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P<0.05). At 10 mm away from the acetabular rim, the average thickness of superior-posterior acetabular wall fluctuated between (8.81±0.67) mm and (13.35±0.89)mm; the average thickness of inferior-posterior acetabular wall fluctuated between (7.02±0.63) mm and (7.66±0.69) mm; the average thickness of acetabular roof fluctuated between (14.46±0.97) mm and (17.05±1.35) mm. Comparatively, the average thickness of superior-posterior acetabular wall was significantly larger than inferior-posterior wall of acetabulum (P<0.05), and the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P<0.01). At 15 mm away from the acetabular rim, the average thickness of superior-posterior acetabular wall fluctuated between (12.08±0.78) mm and (19.84±1.03) mm; the average thickness of inferior-posterior acetabular wall fluctuated between (10.17±0.76) mm and (11.12± 0.77) mm; the average thickness of acetabular roof fluctuated between (23.23±1.12) mm and (26.01±1.53) mm. Comparatively, the average thickness of superior-posterior wall of acetabulum was significantly larger than inferior-posterior acetabular wall (P<0.01), and the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P< 0.01).

CONCLUSIONThe thickness of entire acetabular posterior edge revealed an increasing tendency from inferior-posterior wall to the superior-posterior wall to acetabular roof. And this trend became more obvious with increasing distance away from acetabular rim. Therefore, the superior-posterior acetabular wall could not only maintain the stability of hip joint but also bear loading.

Acetabulum ; anatomy & histology ; injuries ; surgery ; Female ; Humans ; Male

Objective To establish and optimize a sensitive and specific quantitative realtime polymerase chain reaction(PCR)method for detection of hepatitis B virus covalently closed circular DNA(HBV cccDNA)in liver tissue. Methods Specific primers and probes were designed to detect HBV DNA(tDNA)and cccDNA. A series of plasmids(3.44 × 100-3.44 × 109 copies/μl)containing a full double-stranded copies of HBV genome(genotype C)were used to establish the standard curve of real-time PCR. Liver samples of 33 patients with HBV related hepatocellular carcinoma(HCC), 13 Chronic hepatitis B patients(CHB)and 10 non-HBV patients were collected to verify the sensitivity and specificity of the assay. A fraction of extracted DNA was digested with a Plasmid-Safe ATP-dependent Dnase(PSAD)for HBV cccDNA detection and the remaining was used for tDNA and β-globin detection. The amount(copies/cell)of HBV cccDNA and tDNA were measured by a real-time PCR, using β-globin housekeeping gene as a quantitation standard. Results The standard curves of real-time PCR with a linear range of 3.44 × 100 to 3.44 × 109 copies/μl were established for detecting HBV cccDNA and tDNA, and both of the lowest detection limits of HBV cccDNA and tDNA were 3.44 × 100 copies/μl. The lowest quantitation levels of HBV cccDNA in liver tissues tested in 33 HBV related HCC patients and 13 CHB patients were 0.003 copies/cell and 0.031copies/cell, respectively. HBV cccDNA and tDNA in liver tissue of 10 non-HBV patient appeared to be negative. The true positive rate was increasing through the digestion of HBV DNA by PSAD, and the analytic specificity of cccDNA detection improved by 7.24 × 102 times. Liver tissues of 2 patients were retested 5 times in the PCR for detecting cccDNA and the coefficience of variations on cycle threshold (Ct)were between 0.224%-0.609%. Conclusion A highly sensitive and specific quantitative real time PCR method for the detection of HBV cccDNA in liver tissue was established and could be used for clinical and epidemiological studies.

OBJECTIVETo compare different expression profiles of all known chemokine receptors in human hepatocellular carcinoma (HCC) cell lines with different metastasis potentials.

METHODSEighteen pairs of chemokine receptor primers were designed using Premier software. Expression profiles of the 18 chemokine receptors on four HCC cell lines of lower to higher potentials of metastasis (SMMC-7721, MHCC97-L, MHCC97-H and HCCLM6) were analyzed by RT-PCR. Expression of CXCR4 was detected by RT-PCR.

RESULTSExpression profiles of chemokine receptors on four HCC cell lines with different metastatic potentials had significant differences (P < 0.01), in which CCR10, CXCR4 and CXCR6 expressions decreased gradually as the metastatic potential of the cell lines increased. The expressions of CCR3, CCR4, CCR10, CCR12 and XCR1 on HCCLM6 were significantly reduced compared with SMMC-7721 (P < 0.01), whereas the expressions of CXCR1 (P = 0.006) and CXCR5 (P = 0.003) exceeded that of SMMC-7721. Except for CXCR2, CXCR6 and XCR1, most of chemokine receptors on MHCC97-H were expressed differently compared with MHCC97-L (P < 0.05), in which expressions of CCR1 (P = 0.002), CCR2 (P = 0.004) and CCR5 (P = 0.046) exceeded MHCC97-L. CXCR4 was detected only on the positive controls and SMMC-7721 when the template of total RNA was reduced one-half in RT-PCR.

CONCLUSIONChemokine receptors are expressed very differently at mRNA level on HCC cell lines with different metastatic potentials. The different profiles of chemokine receptors in tumor microenvironment and the function of CXCR4 in HCC should be further studied. Our findings have important implications in understanding the relationship between chemokine receptors and the metastatic potential of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Receptors, Chemokine ; metabolism

OBJECTIVETo establish a quick and reliable method to identify Streptococcus oligofermentans, a new species of oral streptococci.

METHODSWith two-step PCR, a pair of the 16S rDNA-specific primers of Streptococcus oligofermentans and a pair of primers of lactate oxidase gene (lox) were used to amplify the gene fragments from the genomic DNAs of 11 strains consisting of 9 species of the pure culture of oral streptococci. Pooled plaque samples from 9 caries-free volunteers were cultured on a selective medium of MSA with erythromycin and tentative strains of Streptococcus oligofermentans were isolated. The isolates were further identified by the two-step PCR and finally confirmed by 16S rDNA sequence analysis.

RESULTSWith the two-step PCR, the two gene fragments were only amplified from the three identified strains of Streptococcus oligofermentans, but not the rest of 8 strains of oral streptococci. Isolates from the dental plaque of caries-free volunteers were identified as Streptococcus oligofermentans by PCR and then further confirmed by 16S rDNA sequence homology analysis.

CONCLUSIONSStreptococcus oligofermentans could be identified by the two-step PCR approach with the specific 16S rDNA primers and lactate oxidase gene primers.

Bacterial Typing Techniques ; methods ; DNA Primers ; genetics ; DNA, Bacterial ; genetics ; DNA, Ribosomal ; genetics ; Mixed Function Oxygenases ; genetics ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; genetics ; Sequence Analysis, DNA ; Streptococcus ; classification ; genetics ; isolation & purification

OBJECTIVETo investigate the construction of the green fluorescent protein (GFP) labeled recombinant adenovirus containing human stem cell leukemia (hSCL) and its distribution and efficiency in mice with interstitial cells of Cajal (ICC) loss.

METHODSThe recombinant adenovirus Ad-GFP/SCL was constructed by Ad-Easy system based on the homologous recombination in bacteria, then 1.6 x 10(9) PFU of recombinant adenoviruses were injected into Balb/c mice with ICC loss via the tail vein. In vivo distribution and efficiency of recombinant adenoviruses mediated hSCL were observed by GFP under the fluorescent microscope at different phases. The expression of SCL gene was measured by RT-PCR method. The damages were observed in different organs by HE staining.

RESULTSThe recombinant adenovirus containing hSCL was quickly constructed by homologous recombination in bacteria using Ad-Easy system. Under the fluorescent microscope, GFP was revealed in heart, lung, liver, spleen, kidney, small intestine and large intestine of mice with ICC loss at different phases. No obvious damages were observed in various visceral organs by HE staining. RT-PCR showed SCL mRNA expression in various visceral organs at different levels.

CONCLUSIONSConstruction of adenovirus vector by the homologous recombination in bacteria is an efficient and time saving method, and a high titer of adenovirus is able to mediate the safe and stable expression of SCL gene in mice with ICC loss. This finding will make further gene therapy in mice with STC possible.

Adenoviridae ; genetics ; metabolism ; Animals ; Constipation ; therapy ; Female ; Genetic Therapy ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Interstitial Cells of Cajal ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; Mice ; Mice, Inbred BALB C ; Recombination, Genetic ; Transduction, Genetic

OBJECTIVETo study the effect of hBcl-2 gene transfer on rat liver against ischemia-reperfusion injury, and explore the feasibility of this approach to reduce ischemia-reperfusion injury in liver transplantation.

METHODSWe constructed the replication-deficient recombinant adenoviruses Adv-EGFP and Adv-Bcl-2 and transfected them into 293 cells and packaged into adenovirus particles for amplification and purification. The empty plasmid vector virus was constructed similarly. Male SD rats were randomized into Adv-Bcl-2-transfected group, Adv-EGFP-transfected group, ischemia-reperfusion group, and sham-operated group, and liver allograft transplantation model was established by sleeve method. In the transfected groups, the recombinant viruses were administered by perfusion through the portal vein, and the ischemia-reperfusion and sham-operated groups received no treatment. Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expressions of bcl-2 in the liver tissue of each group, and at 0, 60 and 180 min after reperfusion, serum AST, LDH, and MDA levels were measured. Histological changes of the liver cells were evaluated by HE staining.

RESULTSBcl-2 mRNA and protein expressions in Adv-Bcl-2-transfected group, as compared with those in Adv-EGFP-transfected group and control group, were significantly increased (P<0.01); the serum levels of AST, LDH and MDA in Adv-Bcl-2-transfected group were significantly lower than those of Adv-EGFP-transfected group and ischemia-reperfusion group (P<0.05 or 0.01). Compared with the sham-operated group, Adv-Bcl-2 treatment group showed lessened edema and vacuolar degeneration of the liver cells without patches or spots of necrosis. In ischemia-reperfusion and Adv-EGFP group, HE staining revealed hepatic lobular destruction and extensive liver cell swelling, enlargement, vacuolar degeneration, edema and occasional focal necrosis.

CONCLUSIONAdv-Bcl-2 transfection can induce the expression of bcl-2 gene to reduce ischemia-reperfusion injury of the liver graft in rats.

Animals ; Genes, bcl-2 ; Liver ; blood supply ; Liver Transplantation ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; pathology ; prevention & control ; Transfection