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.

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.

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.

In recent years, due to the irregular and abused use of glucocorticoid in clinical treatment, the incidence of steroid induced avascular necrosis of femoral head (SANFH) is gradually increasing. TCM for the prevention and treatment of SANFH has received much attention from many scholars. However, due to the lack of the scientific explanation of molecular biology level for its pharmacodynamics mechanism, it is difficult to achieve the purpose of standardized treatment. The discovery of OPG/RANK/RANKL signaling pathway has opened up new shortcuts for the prevention and treatment of bone metabolic diseases. OPG/RANK/RANKL signaling pathway is associated with the pathogenesis of spleen deficiency and phlegm - blood stasis caused by phlegm-blood stasis due to paralysis of SANFH. The treatment efficacy based on the phlegm theory can eventually axial control of the system through the micro-information to express. This article discussed the relevance between the phlegm in the treatment of SANFH and molecular biology mechanism of OPG/RANK/RANKL signal regulation mechanism, and combined the system of bone metabolism regulation mechanism to discuss TCM differentiation of SANFH, with a purpose to provide references for clinical and further study.

ObjectiveTo assess the effect of hemodialysis(HD),hemodiafiltration(HDF) and hemodialysis(HD) and hemoperfusion(HDP) on the quality of life,cognitive function and nutritional status in chronic renal failure(CRF) patients.MethodsAll patients were randomly divided into HD,HDF and HDP group.Mini-mental state examination(MMSE),Wisconsin card sorting test (WCST),SF-36 were measured for cognitive function,quality of life and nutrition indicators test.ResultsAfter 3,6 months treatment,MMSE ( respectively ( 22.62 ±3.85),(25.10 ±2.26),(25.35 ±2.44)and (23.87 ±4.19),(25.10 ±2.26),(27.19 ±2.23)),WCST indicators were statistically significant in the three groups (P ＜ 0.05 ) ;after 1,3,6 months treatment,the PF,RP,BP,GH,SF and nutrition indicators were significant difference too(P＜0.05 ) ; HDP and HDF,HD groups were statistically significant(P＜ 0.05 ).ConclusionHDF and HDP obviously influence on long-term cognitive function,physical health domains and nutritional status in CRF patients,and superior to HD.

Background and purpose:Ovarian cancer-associated ifbroblasts (CAF) are known to promote epithelial malignancy. The chemoattractant cytokine growth-regulated oncogene alpha (Gro-α) secreted from CAF has been reported to mediate the stroma-epithelia interaction in tumor microenvironment, leading to the development of epithelial ovarian cancer, however, the detailed mechanism is unknown.This study was to determine whether Gro-αcould promote ovarian tumorigenesis through activating NF-кB nuclear translocation and VEGF expression in stromal ifbroblasts. Methods:ELISA was used to measure the levels of Gro-αin two cancer-associated ifbroblasts (CAF) and normal ifbroblasts (NF) isolated from high-grade serous ovarian cancer or normal ovarian tissues. CAF conditioned medium (CM) or Gro-αwas used to treat NF, while PS1145, the inhibitor of NF-кB, was used as control. NF-кB subunit p65 and vascular endothelial growth factor (VEGF) were detected by Western blot in cells after treatment. Xenograft tumors from nude mice were generated by injection of CAF, NF, or OVCA429 alone or OVCA429 mixed with CAF or NF, and by injection of OVCA429 mixed with NF cells that were treated with or without CAF-CM or Gro-α, or with NF cells that were treated with CAF-CM or Gro-αplus PS1145. The tumor growth curve was measured and the blood vessel density in xenograft tumor tissues was examined by histopathological analysis. Results:The levels of Gro-αwere 5-6 folds higher in CAF than in NF. Treatment of NF with CAF-CM or Gro-αstimulated the nuclear translocation of NF-кB subunit p65, and the expression of VEGF, but suppressed the expression of thrombospondin 1, the anti-angiogenesis factor, compared with control cells. However, treatment of NF with the NF-кB inhibitor PS1145 reversed these results. The animal assay revealed that CAF stimulated tumor growth stronger than NF, and NF treated with CAF-CM or Gro-α, but not along with PS1145, enhanced xenograft tumor growth through promoting angiogenesis. Conclusion:Ovarian CAF promotes the nuclear translocation of NF-кB and the expression of VEGF through Gro-αautocrine in tumor microenvironment to facilitate angiogenesis and ovarian cancer development.

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.