Objective To study renal involvement in hepatic glycogen storage disease(GSD) in childhood. Methods One hundred and eight patients aged less than 21 years old with type Ⅰa GSD (54 cases), type Ⅲ (29 cases) and uncertain type hepatic GSD (25 cases). Urine analysis, urine albumin, urine protein of 24 h, urine ?_2-MG, BUN, creatinine, Ccr were evaluated. Results Of 108 patients with hepatic GSD, 16 patients (20.8%) had proteinuria proven by urine albumin or urine protein of 24 h, their ages first found proteinuria were 8～15 years. Two 15-year-old patients had proteinuria over 1.0g/24h. Among 72 patients, urine ?_2-MG of 51 cases (70.8%) increased (175～10 623mg/L), and the mean urine ?_2-MG of type Ⅰ a GSD was much higher than that of type Ⅲ GSD, 4138.2 and 1790.1mg/L respectively. Of 91 patients, 10 had renal insufficiency, 1/10 (15-year-old girl) had heavy proteinuria (3.5g/24h), elevated BUN (9.3mmol/L) and Scr(1061 ?mol/L). Five elder patients (11～21 years old) had hematuria with renal colic caused by renal calculus. Conclusions Persistent protenuria, increased urine ?_2-MG, decreased Ccr, and renal stones are common complications of hepatic GSD in childhood. Renal function should be thoroughly evaluated during follow-up.

Objective To study the effects of musk on the expressions of fibroblast growth factor 2 (FGF-2) and epidermal growth factor (EGF) in the rat model of skull bone defect.Methods We constructed the bone defect model by dental drilling into the full skull of 300 SD rats (150 males and 150 females).The model animals were divided with completely random method into model group and drug group,with 150 in each.The two groups were further divided according to drug administration time into 7,14 and 28 d groups,respectively,with 50 in each.The drug group received perfusion of natural musk every day (4.2 mg/100 g) while the model group received perfusion of normal saline of the same volume every day.FGF-2 mRNA and EGF mRNA expressions in skull bone defect were determined using Real-time fluorescent quantitative PCR method.Results EGF mRNA expression at 7 d and 14 d was higher in the drug group than in the model group,but with no significant difference.EGF mRNA expression at 28 d decreased to the lowest level,with a significant difference (P＜0.05).FGF-2 mRNA expression in the drug group reached the highest at 7 d,with a significant difference (P＜0.05),and decreased at 14 and 28 d without significant difference.Conclusion Musk administered at different time points can effectively promote the healing rate of the bone defect area of the rat skull,and the mechanism of this repair is mainly related to the increased FGF-2 mRNA expression and the decreased EGF mRNA expression.

BACKGROUND:Whether the knee is a risk factor for hip fracture is controversial. OBJECTIVE:To investigate the effect and mechanism of knee osteoarthritis in the development of low-energy hip fracture. METHODS:Total y 116 patients with mild traumatic femoral neck fractures and intertrochanteric fractures admitted from October 2014 to January 2015 were enrol ed and retrospectively analyzed. There were 38 males and 78 females. The mean age of onset was (69.2±14.5) years, and al the patients were diagnosed as have simple hip fractures. Anterioposterior and lateral X-ray examination for the hip and knee was performed for each patient. General information, Kel gren-Lawrence grading, scores on Hospital for Special Surgery (HSS), and Western Ontario and McMaster Universities Index of Osteoarthritis were recorded and analyzed. RESULTS AND CONCLUSION:Among the 116 cases, there were 66 cases of femoral neck fractures, 48 of intertrochanteric fractures and 2 of subtrochanteric femoral fractures. According to the Kel gren-Lawrence grading, there were 0 case of grade 0, 21 of grade I, 51 of grade II, 44 of grade III, and 0 of grade IV. Average Kel gren-Lawrence grading was 2.19±0.72 in fractured side and 1.51±0.52 in non-fractured side, and there was a significant difference between the fractured side and non-fractured side (P<0.05). Average score of HSS was 78.4±8.09 in fractured side and 80.2±8.1 in non-fractured side, with statistical difference (P<0.05). These findings indicate that knee osteoarthritis may have some adverse effects in the development of low-energy hip fracture, which can increase the incidence of femoral neck fracture and intertrochanteric fracture.

Objective To investigated the effect of procyanidin (PC) on the expression of cysteine proteinase -3 (Caspase -3) in type 2 diabetes mellitus SD rats with focal cerebral ischemia. Methods Following the random principle, 40 healthy Sprague - Dawley (SD) rats were numbered sequentially and randomly divided to normal rats with focal cerebral ischemia group,type 2 diabetes mellitus SD rats with focal cerebral ischemia group,PC low/ middle/ high -dose groups,with 8 rats in each group. The type 2 diabetes mellitus - MCAO model was set up. The doses of PC for low,middle and high - dose groups were 50 mg/ kg,100 mg/ kg,200 mg/ kg. Immunohistochemistry method was used to measure the activity of Caspase - 3. Results Compared with that in the normal rats with focal cerebral ischemia group[(11. 42 ±2. 52)],the expression of Caspase -3 increased in the type 2 diabetes with ischemia group[(15. 00 ± 2. 38)](t = 2. 17,P < 0. 01). Compared with that in the type 2 diabetes with ischemia group,the expression of Caspase - 3 decreased in the PC groups[(9. 38 ± 2. 00),(7. 71 ± 1. 55),(6. 96 ± 1. 57)](t = 2. 86,3. 13,3. 36,all P < 0. 01),whereby the middle and high - dose groups showed more significant decrease (t = 1. 92,2. 03,all P <0. 01) and with no statistically significant difference between the two groups(t = 1. 13,P > 0. 05). Conclusion PC can decrease the expression of Caspase - 3 protein in type 2 diabetes mellitus SD rats with focal cerebral ischemia, finally may inhibit the apoptosis.

Objective To construct a technical platform for scarless gene modification of Yersinia pestis and to study the functions of its specific genes.Methods The resistance fragment, including upstream and downstream homologous arms of targeted regions, was reamplified by asymmetric PCR.The amplicons were introduced into Y.pestis harboring plasmid pKD46.With the induction of L-arabinose,the recombinant related enzymes: Exo, Beta and Gam, were expressed to guide the homologous recombination.A donor plasmid, pKSI-1, which carried the desired modification fragment flanking by I-SceⅠ recognition sites, was introduced into Y.pestis as the second step of λ-Red recombination with the help of pREDTKI.Results and Conclusion Two mutant strains:△waaA and waaA(△9nt), were successfully constructed for Y.pestis strain 201.Scarless modification introduces no extra modification to the genome, and it is ideal for comprehensive functional genomic studies.

BACKGROUNDOne of mechanisms of carcinogenesis is suppression of cell apoptosis which leads to accumulation of aberrant cells. The aim of this study is to investigate cell apoptosis and COX-2 protein expression in non-small cell lung cancer (NSCLC).

METHODSCell apoptosis, expression of COX-2 and microvessel density (MVD) were detcted in 111 NSCLC samples by TdT-mediated dUTP nick end labeling (TUNEL) technique and immunohistochemical staining.

RESULTSThe positive rate of COX-2 protein expression was 67.6% (75/111), and there were 53 patients with high level cell apoptosis (47.7%). Expression of COX-2 protien was significantly related to TNM stages (P=0.025) and lymph node metastasis (P=0.018). The MVD in NSCLC tissues with positive COX-2 expression was significantly higher than that in negative expression ones (P=0.000). COX model showed that lymph node metastasis (P=0.006) and positive expression of COX-2 protein (P=0.000) were independent prognostic factors of NSCLC.

CONCLUSIONSThe expression of COX-2 protein may suppress cell apoptosis of tumor, and it may serve as a potential marker of prognosis for NSCLC.

Background and purpose: Cancer-associated fibroblasts (CAFs) are known to promote the invasion and metastasis of epithelial cancers. The cytokine IL-6 may mediate the interaction between stromal cells and epithelia in tumor microenvironment to facilitate the invasiveness and metastasis of cancer, however, such mechanism has not been fully covered yet.Methods:We used cervical cancer cell line HeLa as a model for this study. ELISA was used to measure the levels of IL-6 in CAFs and normal ifbroblasts (NFs) isolated from squamous cervical cancer or normal cervical tissues. CAFs conditioned medium or IL-6 was used to treat cervical cancer HeLa cell line. The epithelial-mesenchymal transition (EMT) markers such as N-Cadherin and Vimentin were detected by Western blot in cells before and after treatment. Scratches and transwell chambers were used to test the abilities of cell migration and invasion. Results:The levels of IL-6 were 4-5 folds higher in CAFs than in NFs. Treatment of HeLa cells with CAF conditioned medium or IL-6 upregulated N-Cadherin and Vimentin, but down-regulated E-Cadherin and cytokeratin, compared with control cells, indicating that IL-6 may stimulate HeLa cells to EMT. Further study found that Snail 1, the featured transcription factor for stem cells, was increased along with the enhanced phosphorylation of STAT3. Meanwhile, the migration and invasion of HeLa cells treated with IL-6 or CAF conditioned medium were markedly increased. Conclusion:CAF induces the EMT of cervical epithelial cancer cells through IL-6/STAT3/Snail pathway, which thereby promotes the invasiveness and metastasis cervical epithelial cancer.

Objective To evaluate the imaging characteristics of chondromalacia patellae(CMP)in low-field MR and the value of MR in diagnosis.Methods 37 cases with CMP proved by arthroscopy and/or operation were carefully examined by 0.3 Tesla permanent magnet MR unit.The results of MRI were retrospectively analyzed according to that of arthroscopy and operation to search for the MR imaging manifestations of different degrees of patellae cartilage injuries in CMP.Results According to the golden-standard by arthroscopy and operation as for CMP,the overall diagnostic accuracy of MRI was 79.4%,with 28.6% accuracy in the early phase,92.6% in the late phase of CMP.Conclusion Low-field MR imaging is accurate in diagnosing CMP in the late phase and should be suggested as a useful nontraumatic imaging modality in the assessment of CMP before arthroscopy and operation.

Objective To elucidate functional alterations of glomerular mesangial cell (GMC) contraction and its relationship with diabetic glomerular hyperfiltration. Methods Male Wistar rats were divided into groups of normal control and 2 weeks diabetes induced by STZ. GMC were cultured in the presence of normal glucose (5mM). Meanwhile in comparison GMC obstained from normal rats were cultured in the presence of 30 mM glucose for 48 h in vitro. By using laser scanning confocal microscopy (LSCM) , we measured basal calcium concentration, the changes of calcium level in response to vasoconstrictor agonist angiotensin H (Ang I ),and the effect of protein kinase C (PKC) inhibitor PMAC10 M,24 h)on cytosol calcium of GMCs. Results The creatinine clearance rate (Ccr) was increased with age in diabetic rats, reaching a peak at 2 weeks (P

Objective To study the changes of phosphatidylcholine-specific phospholipase D (PC-PLD) enzyme activity of glomerular mesangial cells (GMCs) in high glucose and the effect of PLD on cytoskeleton. Methods Rats GMCs were cultured in high glucose(30 mmol/L) for 48 hours. PLD enzyme activity was measured by enzyme coupled colorimetry. PKC activity was measured by substrate phosphorylation, and fluorescence intensity of F-actin was observed by FITC-labeled antibody and laser scan cofocal microscopy (LSCM) . Results In high glucose, PLD and PKC activities were obviously elevated, but fluorescence intensity of F-actin was decreased and its cytoskeletal pattern was disassembly. After treatment with inhibtor of PLD, PKC activity was significantly decreased, and intensity and organization of F-actin were recovered. Conclusion Elevation of PLD activity induced by high glucose may influence the cytoskeleton and contraction of CMC through PKC pathway.