OBJECTIVE： To establish a method for detecting glycols in chloramphenicol eye drops METHODS： HPLC method was used with C18 column as stationary phase and methanol－ 0 4％ phosphoric acid（ 55∶ 45） as mobile phase The detection wavelength was 245nm RESULTS： The average recovery was 98 5％ （ RSD=0 86％ ）  CONCLUSION： The method is simple， reliable and suitable for detecting glycols in chloramphenicol eye drops

[Objective]To detect the effect of constant magnetic field on metal ion Co2+,Cr3+ induced TNF-? secreted by human mononuclear cells,and to search a method for prevention and treatment of aseptic loosening. [Methods]CoCl2 powder and CrCl3 powder were dissolved in the asepsis injecting water. Mononuclear cells from human peripheral blood,were taken and cultured with Co2+,Cr3+ ions in different magnetic field of 10Gs,100 Gs,1000Gs for 12,24 and 48 hs. There were nine groups:control group,Co2+ group,Cr3+ group,Co2+andCr3+ with various intensities of constant magnetic field,respectively.ELISA method was applied to detect tumor necrosis factor (TNF-?) in serum via the absorbance (A).[Results]Co2+ and Cr3+ ions stimulated human mononuclear cell to secrete TNF-? (P

Growth hormone (GHRH) and pitutary adenylate cyclase activating peptide (PACAP) are the members of the PACAP/Glucagon superfamily,who are related in both sequence and function.Their stimulation of GH secretion and animal growth is concerned.A series of expression plasmid,pIRES1-GHRH-PACAP (P-G-P),plRESI-GHRH (P-G) and plRESI-PACAP(P-P),were constructed,extracted and purified,then transfected into CHO cell line with Lipofectamine.The expression was examined by RT-PCR,dot-ELISA and Western blotting.The biological activity of expression products was detected in rats.At 8 h after injection of transfection supematant,serum IGF-I concentrations in P-G-P group were significantly higher than that in other groups(P ＜ 0.05).PLGA encapsulating plasmid microspheres were prepared and injected intramuscularly into rabbit legs.Growth behavior and IGF-1 level were measured at day 0,15,30 and 45 after injection.Greater body weights gain and higher serum 1GF- [ levels were observed in three plasmid microsphere injection groups,compared with control group.At day 30,the body weight gain in P-G-P group was greater than saline group (81%,P＜ 0.01),P-G mierosphere group (15%,P＜ 0.05) and P-P microsphere group (7%,P＞ 0.05),serum IGF-I concentration in P-G-P microsphere group showed a 16.68% increase to P-G microsphere (P ＞ 0.05),a 17.14% increase to P-P microsphere(P ＞ 0.05) and a 50.46% increase to control (P ＜ 0.05).These results suggest that co-expression of GHRH and PACAP in one expression plasmid might exert an additive stimulation of GH secretion and growth when delivered into rabbit skeletal muscle with PLGA mierosphere.The results may provide a new approach to regulate animal growth.

This research is to construct a basic medical sciences’experimental teaching system in order to cultivate innovative talents. It is guided by cultivating innovative practical ability and post compe-tence and implements a teaching mode with “five combinations”by integrating teaching resources, strength-ening interdisciplinary combination , integrating curriculum and organ systems , and optimizing teaching modules and experiment content. A preliminary personnel training mode and experimental teaching system have been constructed for innovative talent cultivation, and correspondently a diversified experiment exami-nation system and teaching quality monitoring system have been constructed through teaching practice, which aims at continuously improving experiment teaching quality and talent training quality.

Objective To evaluate the activity of tissue plasminogen activator (tPA) gene transduced endothelial cells (EC) and the cell retention on graft vessel. MethodsKG1 EC were transduced with pseudotyped vectors carrying genes coding for either the wild type tPA or mutant tPA. The supernatants were collecteJPd and assayed for tPA activity in the presence and absence of fibrin. Polytetrafluoroethylene (PTFE) vessel grafts were seeded with EC, and then exposed to an in vitro flow system for 1 h. The number of EC on grafts were counted and the retention of EC were evaluated. ResultsWT5”BZ The tPA activity of the nontransduced EC was (1 5?1 0) IU/ml, while that of wild type tPA gene transduced EC increased to (30.0?8 0) IU/ml, mutant type tPA gene transduced EC increased to (14.1?1 0) IU/ml( P

Objective:To study the antitumor mechanism of W40,a monomer purified from Wedelia prostrate (Hook.et Arn.) Hemsl.Methods:The effects of W40 on the cell proliferative of GLC-82 cells were detected by MTT assay and colony formation assay.The migratory abilities of GLC-82 cells were observed by wound healing assay.Cell apoptosis was evaluated by Annexin V-FITC/PI staining analysis.The levels of apoptosis-relative proteins and cell proliferation-related proteins,such as Caspase-3,PARP,Stat3 and ERK,were detected by Western blotting.Results:MTF assay showed that W40 had a significant cytotoxic effect on non-small cell lung cancer GLC-82 cells.Colony formation assays showed that W40 significantly inhibited GLC-82 cells proliferation.The migration of GLC-82 cells was inhibited by W40 in a dose-dependent manner.Flow cytometry showed that the apoptotic rate increased gradually in a concentration-dependent manner.W40 down-regulated Stat3 as decreasing p-Stat3 and downstream proteins of Bcl-2 and Mcl-1.At the same time,W40 up-regulated the expression of pro-apoptotic protein Bax,and increased the cleavaged Caspase-9,Caspase-3 and PARP.W40 also down-regulated BRAF / MAPK / ERK signal pathway as decreasing p-BRAF,p-MEK and p-ERK.Conclusions:W40 induced apoptosis by inhibiting BRAF / MAPK / ERK and Stat3 signaling pathways.

Objective:To investigate the effect of human growth hormone releasing hormone receptor splice variant type 1 (GHRHR SV1) on the proliferation of human liver cancer HepG2 cells,and to clarify the proliferation effect of GHRHR SV1 on the human cancer cells.Methods:The GHRHR SV1 plasmids were transfected into the human HepG2 cells to construct the HepG2-SV1 cell line.HepG2 group(HepG2 cells),HepG2-empty group(HepG2-pcDNA3.0 cell line) and HepG2-SV1 group(HepG2-SV1 cells) were set up.PCR and Western blotting methods were used to identify the HepG2-SV1 cell line;CCK-8 method was used to detect prolifernation rate of cells;colony formation assay was used to detect the colony formation rate of cells;cell wound healing assay was used to evaluate the migration rate of cells.Results:The PCR and Western blotting results showed the HepG2-SV1 cell line expressed GHRHR SV1 steadily.The CCK-8 results showed that the proliferation rate of the HepG2-SV1 cells in HepG2-SV1 group was higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).The colony formation assay results showed that the colony formation rate of HepG2-SV1 cells in HepG2-SV1 group was 3.5 times higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).The cell wound scratch assay results showed that the migration rate of the HepG2-SV1 cells in HepG2-SV1 group was higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).Conclusion:GHRHR SV1 could increase the proliferation of HepG2 cells.

Objective To explore the molecular mechanism of glucocorticoid (GC)-induced osteoporosis (GIOP). Methods Thirty-two female SD rats after matching body weight were divided randomly into three groups: baseline group (n = 10), control group (n = 11) and GC-treated group (n = 11). The administration time was 9 weeks. Bone mineral density (BMD) was measured with dual energy X-ray absorptiometry. A high resolution micro-CT was used to quantify the densitometric and microarchitectural properties of trabeculae in the proximal metaphysis of right tibia. In situ hybridization histochemistry and immunohistochemistry were used to detect the expression of cannabinoid type 1 receptor (CBI R) in the proximal metaphysis of left tibia. Results At the end of the experiment, whole-body BMD in vivo in the control group [(0. 156±0. 008) g/cm2]was higher than that in the baseline group [(0.147±0.006)g/cm2], while the whole-body BMD in vivo [(0.147±0.006) g/cm2]and total BMD in vitro at femurs in the GC-treated group [(0.220±0.011) g/cm2]was lower than those in the control group [(0. 240±0. 024)g/cm2]. Compared with the baseline group and control group, there was a remarkable decrease in the volumetric BMD, tissue BMD, trabecular number and trabecular connectivity (P<0.05) in the GC-treated group, while there was a significant increase in trabecular separation (P < 0. 05) and trabecular thickness also increased in the proximal metaphysis of tibiae in the GC-treated group. The expression level of CB1R mRNA and protein in osteoclasts in the GC-treated group was markedly higher than that in the baseline group and control group (P < 0. 05). There was a close correlation between the expression level of CB1R mRNA, protein in osteoclasts and some microarchitectural parameters in the proximal metaphysis in the GC-treated group (P<0.05). Conclusions The administration of GC is associated with a decrease in BMD and deterioration in microarchitecture of trabecular bone in rats tibiae. Glucocorticoid may up-regulate the CB1R expression level in osteoclasts and this may be a kind of molecular mechanism of GIOP.

Objective To explore the effect of primary exchange reamed nailing (ERN) and augmentation compression plating (ACP) combined with autogenous bone grafting (ABG) on health-related quality of life in patients with dystrophic or atrophic nonunion of femoral shaft after intramedullary nailing. Methods The study used a prospective study method. Sixty- two patients with femoral shaft nonunion after intramedullary nailing from August 2010 to October 2016 were selected, and the patients were divided into ERN group (group A, 32 cases) and ACP group (group B, 30 cases) by random digits table method. In group A, isthmus nonunion was in 18 cases (56.2％), and non-isthmus nonunion in 14 cases (43.8％); in group B, isthmus nonunion was in 16 cases (53.3％), and non-isthmus nonunion in 14 cases (46.7％ ). The health- related quality of life was compared between 2 groups, including physical component summary (PCS) and mental component summary (MCS) in the- 12- item short form health survey (SF- 12), brief pain inventory- severity (BPI- S) and brief pain inventory- interference (BPI- I). Results Fifty-four patients were followed-up for more than 1 year, and the mean follow-up time was 18.3 (13 to 37) months. All patients successfully achieved bone union, and the mean time was 5.8 (4 to 8) months. Significant improvements in terms of SF-12 PCS and SF-12 MCS score were noted after operation for patients with isthmus nonunion in both groups (t=3.148, 2.156, 2.456 and 2.559; P < 0.05), but there were no significant differences before and after operation in group A with non-isthmus nonunion (P >0.05). At the last follow-up, SF-12 PCS and SF-12 MCS in group B were significantly improved compared with those in group A: (45.2 ± 5.8) scores vs. (33.6 ± 4.7) scores and (48.8 ± 6.5) scores vs. (39.4 ± 5.6) scores, and there were statistical difference (P<0.05); SF-12 BPI-S and BPI-I showed obvious relief: (4.6 ± 2.1) scores vs. (6.2 ± 2.5) scores and (5.2 ± 1.9) scores vs. (6.8 ± 2.7) scores, and there were statistical differences (P<0.05); however there were no statistical difference in SF-12 PCS, SF-12 MCS, BPI-S and BPI- I between 2 groups (P>0.05). Conclusions Compared with ERN combined with ABG, ACP combined with ABG can significantly improve the quality of life in patients with dystrophic or atrophic nonunion of femoral shaft after intramedullary nailing. It has greater advantage on the improvement of health-related quality of life, especially for patients with non-isthmus nonunion.

Objective To analyze the related risk factors of re-nonunion after primary revision for femoral shaft nonunion subsequent to failed intramedullary nailing. Methods A retrospective study was performed in 61 patients with femoral shaft nonunion subsequent to failed intramedullary nailing from June 2008 to June.All patients were divided into re-nonunion group(22 cases)and non-re-nonunion group (39 cases) according to diagnostic criteria of bone re-nonunion. Univariate analysis was used to analyze 14 factors that may lead to the occurrence of re-nonunion after revision for femoral shaft nonunion subsequent to failed intramedullary nailing including age, gender, body mass index (BMI), smoking, alcohol abuse, injury reason, fracture types, intramedullary nail types, locking screws technology for intramedullary nail, bone nonunion sites, bone nonunion time, pathological types of bone nonunion, primary revision methods and autologous bone graft or not, and multi-factor logistic regression analysis was performed on the factors showing a significant difference. Results Univariate analysis showed significant difference in smoking (χ2= 6.564, P = 0.036), BMI (χ2= 6.783, P = 0.021), bone nonunion sites(χ2=7.316,P=0.011),primary revision methods(χ2=8.069,P=0.003)and autologous bone graft or not(χ 2=6.668,P=0.027).Logistic regression analysis showed that primary revision methods(OR=1.027,95% CI 0.028-0.463,P<0.05)and autologous bone graft or not(OR=1.024,95% CI 0.006-0.363, P < 0.05) were independent risk factors for re-nonunion after revision of femoral shaft nonunion subsequent to failed intramedullary nailing. Conclusions Primary revision methods and autologous bone graft or not are independent risk factors for re-nonunion after revision of femoral shaft nonunion subsequent to failed intramedullary nailing.By strictly controlling the surgical indications and combining with autogenous bone grafting,it is possible to reduce the occurrence of nonunion after primary revision of the femoral shaft nonunion subsequent to failed intramedullary nailing.