Objective To investigate the gene therapy of hypoparathyroidism (HPT) with hemopoietic stem cell s(HSCs) integrated with parathyroid hormone (PTH) gene.Methods Human PTH gene was amplified by polymerase chain reaction from pcDNA 3.1 -PTH templates and inserted into murine stem cell virus (MSCV) vectors. The plasmids recombinated with PTH gene were transfected into PA317 packaging cells and screened for positive clones with Geneticin. The condensed recombinant retroviruses were obtained and used to infect HSCs coming from umbilical cord blood. The infected HSCs were injected into the mice suffering from HPT. The change of symptoms and serum level of PTH and calcium in each group of mice was observed.Results Human PTH gene was proved to be inserted into MSCV vectors and the titres were up to 2?10 7 colony forming unit (CFU)/ml in condensed retroviral solution. The secretion of PTH reached 15 ng per 10 6 cells per 48 h. The wild type viruses were not detected by PCR amplification. The mice suffering from HPT recovered quickly and the serum level of calcium and PTH remained normal for about 3 months after the HSCs recombinated with PTH gene were injected into them. In the groups of mice subject to the injection of the recombinant retroviruses containing PTH gene, the serum level of PTH and Calcium was significantly increased in a short time, and then declined slowly. And the mice took on the manifestation of hypoparathyroidism again. The mice only subject to the implantation of HSCs were all dead within 20 days.Conclusion A long term therapeutic effect could be obtained by infusing the HSCs which were integrated with PTH gene into the mice suffered from hypoparathyroidism.

BACKGROUND:Endothelial progenitor cells are recruited into local vascular injury under the injury-induced stimulation, and then differentiate into mature endothelial cells that are thereby involved in angiogenesis and endothelial repair. OBJECTIVE:To investigate whether endothelial progenitor cells can al eviate renal injury and improve renal function of ischemia/reperfusion injury (I/R) rats. METHODS:Peripheral blood samples extracted from Sprague-Dawley rats were used to isolate and culture endothelial progenitor cells using density gradient centrifugation. Twenty-two male Sprague-Dawley rats were randomized to three groups:I/R group, normal control group, and endothelial progenitor cells group. In the I/R and endothelial progenitor cells groups, the right kidney was removed and the renal artery and vein of the left kidney were occluded for 40 minutes to establish I/R models in the rats, and then endothelial progenitor cells (5×109/L, total y 1 mL) or solvent was transplanted via the artery of the left kidney into the left kidney. In the normal control group, the experimental procedure was same as that in the I/R group except for occlusion of the artery and vein of the left kidney. Renal and blood samples from three groups were col ected at day 1 after operation. Peripheral blood CD34 and vascular endoethelial growth factor receptor 2 expressions were determined using flow cytometry and immunofluorescence methods, serum creatinine and urea nitrogen were tested, and immunohistochemistry observation was used for CD34 observation. RESULTS AND CONCLUSION:Compared with the normal control group, serum creatinine and urea nitrogen levels were significantly increased, and tubulointerstitial CD34 expression was decreased in the I/R group (P<0.05). Endothelial progenitor cells treatment largely decreased the levels of serum creatinine and urea nitrogen, and increased CD34 expression (P<0.05). These findings indicate that transplantation of endothelial progenitor cells contributes to renal protection in I/R rats.

Objective With the improvement of technology and strategy , reduced field-of-view diffusion weighted imaging ( rFOV DWI) has provided more robust imaging .This article was intended to compare the clinical utility between rFOV DWI and con-ventional DWI in the diagnosis of prostate cancer . Methods Retrospective analysis was made on 58 prostate cancer patients under-going verification of biopsy or surgery after 3.0 T MRI rFOV DWI and regular DWI examinations in Nanjing General Hospital of Nan-jing Military Command from August 2014 to September 2015 , among whom prostate cancer and benign prostatic hyperplasia were 31 cases and 27 cases respectively .Independent grading was made by 2 radiol-ogists who were unaware of any clinical data and sequences in the as-pects of distortion on DW images , lesion conspicuity on DWI ( b=1000 ) and ADC images according to a 4-point Likert scale .rFOV and conventional DWI sequences were acquired using b =400,1000,0s/mm2.The IVIM double exponential model software was applied to generate ACD value, slow dispersive coefficient(D value) and parameterized image of volume fraction of fast dispersion (f value).Se-quence analysis was made in suspected peripheral tumor regions of prostate cancer patients and and healthy peripheral zones of patients with benign prostatic hyperplasia respectively to get ACD , D and f values .The ratio values rADC , rD and rf were attained compared with obturator internus .ROC analysis was made on the efficacy of different parameters of two sequences in prostate cancer . Results Image qualities of rFOV DWI were rated superior to conventional DWI (P<0.001,Wilcoxon Signed-Rank Test).Compared with con-ventional DWI, ADC value (0.728 ±0.094 vs 0.789 ±0.116, P<0.05)and D value(0.622 ±0.077 vs 0.651 ±0.099, P<0.05)of peripheral tumor regions in rFOV DWI were lower and the difference was statistically significant , whereas no significant difference was found in f value(P=0.932).No difference was found in area under ROC curve of f value between rFOV DWI and traditional DWI (0.853 vs 0.767, P=0.283). Conclusion The rFOV DWI sequence yielded can efficiently improve image quality , which is of important value to the clinical diagnosis of prostate cancer .

Purpose To observe the mutation of K-ras gene and expression of Fascin-1 protein in CRC tissues and their relationship with clinical pathological features, and then to analyze the correlation between mutation of K-ras and expression of Fascin-1. Methods In 86 cases of CRC tissues, K-ras mutation was detected by DNA sequencing analysis, and Fascin-1 expression was detected by im-munohistochemical method. Results In CRC tissues the mutation rate of K-ras was 34. 88%, the expression rate of Fascin-1 was 60. 47%. The mutation rate of K-ras in lymph node metastasis group was higher than that of without lymph node metastasis group, and that in distant metastasis group was higher than that of without distant metastasis group(P<0. 05). The expression rate of Fascin-1 in serosa invasion group was higher than that of without serosa invasion group, and that in lymph node metastasis group was higher than that of without lymph node metastasis group, and that in distant metastasis group was higher than that of without distant metastasis group (P<0. 01). There was a correlation between the mutation of K-ras gene and the expression of Fascin-1 in CRC tissues (rp =0. 236, P<0. 05). Conclusions The CRC tissues with mutation of K-ras are more likely to metastasize and the CRC tissues with expression of Fascin-1 are more likely to invade serosa and metastasize. The CRC tissues with mutation of K-ras are more likely to express Fascin-1.

Objective To investigate 4 variants single nucleotide polymorphisms (SNPs) of 5-lipoxygenase-activating protein(ALOX5AP) in lipoxygenase pathway and in cytochrome P450 pathway as susceptibility genes for stroke in a southeastern Chinese population,and evaluate the associations between susceptibility genes and cerebral infarction,to find whether gene-gene interactions increase the risk of cerebral infarction.Methods By case-control study,two hundred and ninety-two patients with cerebral infarction and 259 healthy control subjects were included.Eight variants in 5 candidate genes were examined for stroke risk,including the SG13S32 (rs9551963),SG13S42 (rs4769060),SG13S89 (rs4769874),and SG13Sl14 (rs10507391) variants of the ALOX5AP gene,the G860A (rs751141) variant of the soluble epoxide hydrolase (EPHX2) gene,the A1075C (rs1057910) variant of the CYP2C9 *2 gene,the C430T (rs1799853) variant of the CYP2C9* 3 gene,and the A6986G (rs776746) variant of the CYP3A5 gene.Gene-gene interactions were explored using generalized multifactor dimensionality reduction (GMDR)methods.Results There were no statistically significant differences in the frequencies of the genotypes of the 8 candidate genes.The GMDR analysis showed a significant gene-gene interaction between SG13S114 and A6986G,with scores of 10 for cross-validation consistency and 9 for the sign test (P =0.011).These genegene interactions predicted a significantly higher risk of cerebral infarction (adjusted for age,hypertension,and diabetes mellitus;OR =1.804,95％ CI 1.180-2.759,P =0.006).Conclusions A two-loci gene interaction confers significantly higher risk for cerebral infarction.The combinational analysis used in this study may be helpful in the elucidation of genetic risk factors for common and complex diseases.

PurposeTo evaluate the existence and extent of magnetic resonance(MR) artifacts caused by frequently used metallic dental materials and to compare the influence of different MRI sequences on artifacts.MethodsA total of 22 kinds and 25 metallic dental samples were tested with 1.5 T MR imager and gradient-echo sequence. Spin-echo and fast spin-echo were added to parts of these samples. Results Of all the 25 metallic dental samples, 11 including gold, amalgam, and silver point did not produce artifact. Titanium alloy and porcelain product fused in metal had mild artifacts. Whereas the remaining 12 samples such as the retention pin and pivot pin showed severe artifacts. Artifacts produced by retention pin, nickel chromium crown and so on were less severe on fast spin-echo. ConclusionsAttention should be paid to some of the metallic dental materials, which could cause severe MR artifacts and image degradation, when undergoing face,jaw and head MR examination. Artifacts can be alleviated by using proper metallic materials or choosing proper imaging sequence and parameters.

Objective Objective To analysis the clinical curative effect on elderly patients with choledocholithiasis by endoscopic retro-grade cholangiopancreatography ( ERCP) and endoscopic sphincterotomy ( EST) and discuss its clinical significance. Methods 256 patients with choledocholithiasis were collected for diagnosis and treatment by ERCP. The stone characteristics was mastered by ERCP and the pa-tients were treated with EST. Results Patients with choledocholithiasis were successful in 249 cases (97. 27%) for ERCP intubation and 239 cases (93. 36%) for stone remove. After EST,the stones were removed successfully at one time in 199 cases (83. 26%),2 times in 40 cases (16. 74%). 7 patients (2. 73%) with multiple diverticulum and papillary position poor by intubation failure were changed over to op-eration treatment. After ERCP,patients were performed endoscopic nasobiliary drainage (ENBD) 64. 44% (154/239). Pull out the drain-age tube of 90. 91% (140/154) of the patients after stones disappeared which was proved by angiography for 3~5 days. 14 cases (9. 09%) were performed second times to remove the stones due to the presence of residual stones. Postoperative complications occured in 15 cases (6. 02%) including 9 cases of acute pancreatitis and 6 cases of infection of biliary tract, and they were cured after 1 week of corresponding treatment. Transient increase of serum amylase occurred in 39 cases, and all of them recoveried after 3 days without special treatment. Con-clusion The results showed that ERCP ( or EST) were well tolerated by elderly patients with choledocholithiasis,and they were of obvious curative effect and quick recovery,which is worthy of promotion and application.

The source of seed cells has always been the major problem in cartilage tissue engineering.Bone marrow stromal cells (BMSCs) have gradually become an optimal source of seed cells for cartilage engineering due to their high proliferative potential,multi-lineage differentiation potential and easiness to be obtained with minute trauma.The great challenge is how to get abundant BMSCs with a high purity and how to induce them in vitro into chondrogenic phenotype.This review aims to discuss the various strategies that can induce BMSCs chondrogenic differentiation in vitro so as to offer beneficial reference for constructing cartilage with BMSCs as seed cells.

Articular cartilage defects are commonly found in clinics. It is always a great challenge for the repair of cartilage defects due to the limited self-regeneration after injury. Tissue engineering,a newly emerging biotechnique to regenerate cartilage with chondrogenic potential cells and biodegradable scaffolds in vivo and in vitro,provides a promising method to solve the challenge in cartilage defects repair. To regenerate cartilage with favourable structure and function,it is essential to gain a deep insight into the biochemical structure,biomechanical property and their relationship of articular cartilage. This article gives an introduction to the biochemical structure,biomechanical property and their relationship of both native and tissue-engineered cartilage.

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