BACKGROUND:Large bone defect caused by various reasons has been a difficult problem in clinical practice. To establish a standard experimental animal model of critical bone defects has vital significance for evaluating the efficacy of bone osteogenesis using various materials and techniques.
OBJECTIVE:To establish the rabbit model of parietal critical bone defects and to determine the diameter of the critical defects of parietal bone in limited time.
METHODS:10 New Zealand white rabbits were selected. The skul seam was treated as the boundary. Four ful-thickness round defects of bone in the parietal bone were made, with diameters of 4, 5, 6 and 7 mm, so as to establish rabbit models of parietal critical bone defects. Gross anatomical observation, X-ray and cone beam CT were used to determine the bone density in the new bone defect area. The healing of bone defects was evaluated by histological examination.
RESULTS AND CONCLUSION:After 12 weeks, the 4 mm group showed high bone healing capacity significantly, and part of the bone bridge had been connected completely. Quantitative analysis of bone mineral density revealed that gray value at defect site and trabecular bone area at the same magnification and the same vision in the 4 mm group were significantly higher than the other three groups (P<0.001). Only a smal amount of new bone in the periphery of bone defects appeared in the 5, 6 and 7 mm groups. The center of defect site was mainly fil ed by fibrous connective tissue. The results confirmed that this study successful y established rabbit models of parietal critical bone defects. During the 12 weeks of observation, bone defects with a diameter of ≥ 5 mm could not be self-healed, which was conformed to the criteria of critical defects of bone, and could be used as a reference value for critical parietal bone defects of a rabbit.

Prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), plasma fibrino-gen (FIB), platelet aggregation rate and blood clots-fibrinolytic dynamic figure were taken as indexes in the evalua-tion of anticoagulant activity in vivo of active component F2-2 from Eupolyphaga seu Steleophaga. After 5 days of hypodermic injection of adrenaline, the rat model of acute blood stasis was established. Indexes were determined af-ter the model rats were treated with an intragastric administration of F2-2 for 9 days. The results showed that com-pared with the model group, PT/APTT was prolonged, FIB content was decreased, platelet aggregation rate and the largest of blood coagulation were declined after 9 days of intragastric administration in the model group. However, there was no difference on TT. It was concluded that the anticoagulant component F2-2 separated from E. seu Steleophaga showed favorable anticoagulant activity in vivo. However, its mechanism remained unknown and request-ed further researches.

An adult patient with hypophosphatasia caused by compound heterozygous mutations in alkaline phosphatase,liver /bone /kidney(ALPL)gene was investigated through comprehensively reviewing the medical history and clinical records of the proband and her family members in order to better understand the disease.The proband and her older sister had mild decreased serum alkaline phosphatase level accompanied with frequently nontraumatic fractures at limbs and all the teeth fell off at the age of 20 and 7, respectively.Both of them carried a missense mutation c.407G>A(p.Arg136His)in exon 5 and a deletion mutation c.1318_1320delAAC(p.Asn440del)in exon 12 simultaneously.Other four family members were p.Arg136His mutation carriers and two members were p.Asn440del mutation carriers.We found that p.Asn440del mutation was associated with the oral disorders.In this family, compound heterozygous manifested more serious symptoms, while heterozygous showed relatively mild symptoms.In addition, it is necessary to differentiate it from primary osteoporosis and other diseases of disturbed bone mineralization.

Objective To compare the osteogenesis effect of platelet-rich fibrin (PRF) and platelet-rich plasma (PRP) and investigate the methods of repairing bone defect with PRF.Methods Four defects measuring 7 mm in diameter were made in the parietal bone of 16 New Zealand white rabbits.The defects named A,B,C,and D and were filled with PRF,PRF-mixed Bio-Oss (BO),PRP-mixed BO,and PRP separately.Every four rabbits were sacrificed at postoperative 2,4,8,and 12 weeks and defects were examined grossly,radiographically,and histologically.Besides,bone mineral density and bone trabecular area were determined and expressed as gray-scale values.Results Newly regenerated bone appeared at all defect areas at postoperative 2 weeks.Thereafter,more bone formations were observed over time and area B demonstrated the best bone healing followed by area C,A,and D in succession.Bone trabecular area in areas A,B,C,and D was 10.95 ± 0.58,15.45 ± 0.79,10.22 ± 0.43,and 6.58 ± 0.64 at postoperative 2 weeks with significant differences in pair comparison (F =22.869,P ＜0.01),followed by some increase at postoperative 4 and 8 weeks.Whereas,bone trabecular area in areas A,B,C,and D increased largely at postoperative 12 weeks (35.09 ± 0.58,59.44 ± 0.60,50.75 ± 1.56,and 30.94 ± 1.19) and showed significant difference when compared in a pair (F =1 002.904,P ＜ O.01).Conclusion PRF is superior to PRP in promoting bone formation,but a much better effect of PRF/BO composite is observed in bone repair.

Objective To investigate the association of estrogen receptor-? (ER-?) and vitamin D receptor (VDR) gene polymorphisms with peak bone mass in Shanghai women. Methods The ER-? PvuⅡ and XbaⅠ genotypes and VDR ApaⅠ genotypes were determined by PCR-RFLP in 515 unrelated healthy women aged 19-40 years of Han nationality in Shanghai. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry. Results Frequencies of ER-? PvuⅡ genotype PP, Pp and pp were 13.2%, 49.3% and 37.5% respectively. Frequencies of ER-? XbaⅠ genotype XX, Xx and xx were 4.7%, 40.4% and 54.9% respectively. Frequencies of VDR ApaⅠ genotype AA, Aa and aa were 5.8%, 41.9% and 52.3% respectively. Hardy-Weinberg equilibrium was evident for both ER-? and VDR gene polymorphisms. No association was found between ER-? PvuⅡ and XbaⅠ genotypes and BMD of various sites in women. Only a significant association was found between VDR ApaⅠ genotype and BMD at L 1-4(P

OBJECTIVETo study the injection of NKG5SV gene to inhibit growth and metastasis of hepatocellular carcinoma (HCC).

METHODSNKG5SV gene was inserted into retroviral vector pLXSN by normal methods. LacZ gene was used as control. LCI-D20 tumor together with saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was subcutaneously inoculated to the nude mice. Tumor formation rate and tumor size were noted 35 days after inoculation. LCI-D20 tumor was inoculated subcutaneously. Saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was intratumorally injected respectively 10 days after inoculation. Tumor growth was observed 35 days after inoculation. Liver cancer was resected 22 days after intrahepatic inoculation. Saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was respectively injected at incisal margin or intraspleen. Mice were killed 35 days after inoculation to observe tumor recurrence at incisal margin, intrahepatic metastasis and extrahepatic metastasis.

RESULTSTumor formation rate and tumor diameter(cm) were 1.76 +/- 0.11, 1.51 +/- 0.34, 0.33 +/- 0.04 in the control group, LacZ group, NKG5SV group respectively when tumor and different cDNA were inoculated together. Tumor diameter(cm) and weight(g) were 0.87 +/- 0.08, 0.83 +/- 0.05, 0.26 +/- 0.04; 0.43 +/- 0.06, 0.38 +/- 0.04, 0.08 +/- 0.06 in the control group, LacZ group, NKG5SV group respectively when different cDNA were injected into the LCI-D20 tumor. Sites with extrahepatic metastasis nidi, incisal margin recurrence tumor size(cm), intrahepatic metastasis nidi, metastasis involved hepatic lobes in the control group, LacZ group, NKG5SV group were 4.25 +/- 1.48, 4.25 +/- 1.04, 0.63 +/- 0.51; 1.51 +/- 0.27, 1.35 +/- 0.17, 0.81 +/- 0.17; 2.50 +/- 1.41, 2.38 +/- 1.06, 1.25 +/- 0.71; 2.13 +/- 0.99, 2.00 +/- 0.75, 1.38 +/- 0.74 respectively when NK cells were injected at incise margin. They were 4.38 +/- 1.85, 4.25 +/- 1.48, 1.00 +/- 0.75; 1.13 +/- 0.23, 0.97 +/- 0.29, 0.76 +/- 0.16; 2.50 +/- 1.41, 2.05 +/- 1.12, 0; 2.13 +/- 0.83, 1.75 +/- 0.88, 0 respectively when NK cell were injected intrasplenicly.

CONCLUSIONSNKG5SV gene can inhibit HCC growth and postoperative metastasis and recurrence.

Animals ; Antigens, Differentiation, T-Lymphocyte ; Cell Division ; drug effects ; Genetic Therapy ; methods ; Genetic Vectors ; administration & dosage ; genetics ; Humans ; Injections ; Liver Neoplasms, Experimental ; genetics ; pathology ; therapy ; Male ; Mice ; Mice, Nude ; Neoplasm Metastasis ; prevention & control ; Receptors, Immunologic ; genetics ; physiology ; Tumor Cells, Cultured ; Xenograft Model Antitumor Assays

Objective:To investigate the effect of deviation from the isocenter point on the quality of CT images at the same radiation dose.Methods:A 160-layer CT scanner was used to scan the phantom at isocenter and deviations of 3, 6, 9, 12 and 15 cm. CT was performed with the following parameters: 120 kVp; 400 mAs; slice thickness, 1 mm; and slice increment, 1 mm. Images were reconstructed using the filtered back projection algorithm. Noise power spectrum (NPS), task transfer function (TTF) and detectability index (d′) were measured. NPS peak was used to quantify the noise magnitude and TTF 50% was used to quantify the spatial resolution. NPS, TTF and d′ were compared using one-way ANOVA. Results:The NPS average spatial frequency, spatial resolution and d′ values gradually decreased as the offset distance increased and the amount of noise increased. NPS peak at isocenter and deviations of 3 cm, 6 cm, 9 cm, 12 cm and 15 cm were (94.31±1.48), (104.25±1.46), (131.44±1.96), (171.86±1.91), (224.05±1.37), (286.51±2.09)HU 2·mm 2, respectively ( F=37 241.91, P<0.001). And d′ values of 2 mm low-contrast lesions were 3.51±0.06, 3.31±0.04, 3.01±0.04, 2.59±0.06, 2.21±0.03, 1.88±0.03, respectively. The reduction in spatial resolution was more pronounced for high contrast, and the d′ values decreased to a similar extent for various types of lesions. The noise was increased by about 82%, the high contrast spatial resolution was decreased by about 12%, and the d′ value was decreased by about 26% at 9 cm from the isocenter point (all P<0.05). The noise was increased by about 204%, the high contrast spatial resolution was decreased by about 27%, and the d′ value was decreased by about 45% at 15 cm from the isocenter (all P<0.05). Conclusion:The CT image quality was decreased with the increase of the offset distance from the CT isocenter point. The image quality was severely compromised at offset distances greater than 9 cm.

Large cell neuroendocrine carcinoma of bladder is a rare malignant tumor with high degree of malignancy, strong invasiveness and poor prognosis. We reported a case of a 56-year-old man who underwent transurethral resection of bladder tumor because of bladder mass. Postoperative pathology revealed large cell neuroendocrine carcinoma of the bladder with urothelial carcinoma. Radical cystectomy was performed after postoperative chemotherapy, and there was no recurrence after 3 months of follow-up.

Objective To investigate the application value of the fluorescence quantitative PCR(qPCR)melting curve method in the detection of clarithromycin(23S rRNA)and quinolone(gyrA)resistant genes of Helicobacter pylori(Hp)in fecal samples.Methods A total of 1 176 untreated patients who underwent gastroscopy and were Hp positive proved by rapid urease test(RUT)were enrolled in the study.Their gastric mucosal and fecal samples were collected.The E-test method was used to analyze the clarithromycin and quinolone resistant phenotypes of Hp in gastric mucosal samples.The qPCR melting curve method was used to detect the clarithromycin and quinolone resistant genotypes of Hp in fecal samples.The consistency of the results obtained by the two methods was evaluated by the Kappa test.In addition,the nucleic acids were extracted from the fecal samples with Hp positive,and the Hp 23S rRNA and gyrA resistance mutation genes were detected by the qPCR melting curve method and Sanger sequencing,respectively.The consistency of the results obtained by the two methods was compared.Results In the study of clarithromycin resistance,a total of 934 valid samples were obtained.Among them,453 samples had positive resistance phenotype and 481 had positive resistance genotype,with a positive consis-tency rate of 93.38%(95%CI:90.70%～95.32%).In the study of quinolone resistance,a total of 909 valid samples were obtained.Among them,426 samples had positive resistance phenotype and 413 had positive resistance genotype,with a positive consistency rate of 86.85%(95%CI:83.31%-89.74%).In the comparative study,986 valid samples were detected for Hp 23S rRNA gene.Among them,514 samples were resistance positive detected by the qPCR melting curve method and 509 by Sanger sequencing,with a positive consistency rate of 96.27%(95%CI:94.24%-97.60%).Similarly,895 valid samples were detected for Hp gyrA gene.Among them,422 samples were resistance positive detected by the qPCR melting curve method and 405 by Sanger sequencing,with a positive consis-tency rate of 95.80%(95%CI:93.38%-97.36%).Conclusion The qPCR melting curve method can detect Hp 23S rRNA and gyrA in fecal samples,which has certain clinical application value for predicting the resistance of Hp.

Objective Proteus syndrome is a rare disease. The aim of the present study was to analyze the clinical characteristics and gene mutations of Proteus syndrome with a case report and relevant literature review. Methods Clinical data of the patient with Proteus syndrome were collected in detail and biochemical measurements and radiological examinations were conducted. Tissues from phalanges with lesions were obtained to extract DNA, and Sanger sequencing of AKT1 gene was carried on. The pathogenic mutation was further tested in peripheral blood samples of the patient, his parents and 250 healthy volunteers. Orthopaedic surgery was performed on the affected limbs of the patient. Results The patient was presented with progressive overgrowth of the right extremity, scoliosis, cerebral connective tissue nevus and lower extremity venous. A heterozygous mutation of AKT1 gene (c. 49G>A) was identified in DNA extracted from the affected bone tissue of the patient, but not be found in genomic DNA of peripheral blood samples from the patient, his parents and 250 healthy volunteers. Movement function of the affected limb improved significantly after the operations. Conclusions The prominent features of Proteus syndrome are overgrowth of one extremity and cerebral connective tissue nevus. A mosaic somatic mutation of AKT1 gene is one of the pathogenic mutations for Proteus syndrome, and orthopedic surgery may be a good way to improve symptoms of the disease.