Objective To evaluate the effect of human bone morphogenetic protein 2 (hBMP-2)/Bone Mesenchymal Stem Cells (BMSCs)/demineralized bone matrix(DBM) on repairing rabbits’femoral head after necrosis and to explore the new treatments for femoral head necrosis. Methods Femoral head necrosis models was established by clinical core decom?pression combined with liquid nitrogen frozen. Then, animals were randomly devided into 4 groups (n=12 per group):Group A were not implanted anything as control group, Group B were implanted with DBM. Group C were implanted with hBMP-2/DBM. Group D were implanted with hBMP-2/BMSCs/DBM. Four rabbits from each group were sacrificed at 4,8 and 12 weeks after surgery to evaluate the the repairing effect of Osteonecrosis of the femoral head (ONFH) through X-ray examina?tion, observation of the specimen and HE staining. Results X-ray revealed defect of femoral head in Group A without clear bone formation. There is a little fibrous hyperplasia and no obvious osteogenic response. By contrast, the femoral head defect areas became fuzzy in group B, group C and group D with new bone trabeculars. And the regenerate phenomenons of group D were significantly better than that of group B and group C of the same time point. As to the Lane-Sandhu X Ray scores, it is lower in group A than that in group B;It is lower in group C than that in group D(P<0.05). There is no statistical difference between Group B and Group C. General observation of the specimen revealed that the femoral head of group A collapsed with drilling holes. The femoral heads of group B and group C showed no collapse but the drilling holes existed. Femoral head in group D was not collapsed and the drilling holes disappeared. HE staining showed that bone trabeculars became ne?crotic and fragmented in Group A with a lot of air trapped cells. There were newborn immature bone trabeculars and osteo? blasts in group B and group C. Group D were of large number of bone cells, fat cells, and newborn mature bone trabeculars. The ratio of empty lacuna is higher in Group A than that in Group B;it is higher in Group C than that in Group D(P<0.05). Conclusion hBMP-2/BMSCs/DBM can induce BMSCs differentiation into osteoblasts after being implanted. It has good re?pairing effect on ONFH with good application prospect.

Objective To study blood type B antigen elimination with α-galactosidase in human liver tissue,and discuss the feasibility of blood type conversion in human liver.Methods The liver specimens from patients with blood type B in liver transplantation were collected,and an in vitro liver perfusion model was established.The in vitro livers were perfused with UW solution +/-α-galactosidase.The effect of enzyme in B antigen of human liver were analyzed by immunofluorescence.Results With UW solution containing α-galactosidase to perfuse the in vitro livers,immunohistochemistry showed the level of blood type B antigen in liver was significantly reduced after hypothermic perfusion and preservation.The B antigen level in 1 h perfusion was reduced to approximate 58％ of this figure prior to perfusion,in 2 h was 10％,and in 4 h was 4％.Among the different intervals,the blood group antigen levels showed significant differences (P ＜ 0.05).In the control group,the blood group antigen levels showed no obvious change on statistical analysis.Conclusions α-galactosidase was effective to clear blood type B antigen in isolated liver tissue.In the experimental group,Although the B antigen did not fall to a undetectable level,liver blood type conversion from B→O remains a promising potential which has been meaningful for related researches on blood type conversion of human organs.

Objective To investigate the role of interferon regulatory factor-1 (IRF-1) in liver ischemia/reperfusion (IR) injury and its underlying mechanism,and identify effective managements in alleviating liver IR injury.Methods Three groups of mice models with liver IR injury were well established,including control group (S),warm liver IR injury group (IR) and recombinant IRF-1 group (IRF-1).The levels of mRNA and protein,liver function and pathological changes of liver tissue were detected in group S and group IR.Additionally,the marker of IRF-1,p-Stat1,p-P38,PARP1 and Caspase-3 were measured and PCNA expression was determined in group IR and group IRF-1 mice with 6-hour liver IR injury.Results IRF-1 mRNA and protein and the levels expression of proteins were significantly elevated with peak occurred after 6-hour IR injury,which was statistic difference compare to the group S (t2h =-3.512,t6h =-4.247,t12h =-4.088,t24h =-3.851;P ＜ 0.05).Serum ALT and AST of mice detected in group IR were higher than group S at all endpoints (tALT =4.931,4.592,4.277,4.809;tAST =4.980,4.617,4.336,4.915;P ＜ 0.05).Furthermore,pathological damage change was more distinct compared with group S.The elevated levels of IRF-1,p-Statl,p-P38,PARP1 and Caspase-3 and decreased PCNA expression were determined in mice models with recombinant IRF-1 intervention.Conclusion IRF-1 expression could be closely correlated with liver IR injury,and its underlying mechanism may be attributed to activation of JNK MAPK protein and inhibition of PCNA expression.

Objective To observe the specific changes of content of cyclic guanosine monophosphate (cGMP) around meridian skin and specific influences of electroacupunture Neiguan (PC6) on cGMP of rats with acute myocardial ischemia (AMI). Methods With 6 male Wistar rats as normal group, 18 rats were made AMI model and divided into model group, high-frequency group and low-frequency group (n=6). The high-frequency and low-frequency group were given electroacupuncture in bilateral PC6 with 100 and 2 Hz for 20 min respectively. The content of cGMP in skin around PC6, Ximen (PC4), Tianquan (PC2), Waiguan (TE5) and Zusanli (ST36) was detected by using enzyme-linked immunosorbent method after treatment. Results In model group, cGMP content increased significantly around PC6, PC2, TE5 and ST36 compared with normal group (P <0.05,P <0.001). After the treatment with high-frequency or low-frequency electroacupunture in PC6, cGMP content decreased significantly in skin around PC6, PC2, TE5 and ST36 compared with model group (P <0.05, P <0.001). Conclusion High-frequency electroacupunture or high-frequency electroacupunture used in PC6 can protect rats from the injury of AMI. The mechanism may be related to the decrease of cGMP content around the acupoint areas in heart-related meridian.

BACKGROUND: Both lentiviral vector and adenoviral vector are considered as good vectors for gene mediation, and their differences in transferring bone morphogenetic protein 2 (BMP-2) into rabbit bone marrow mesenchymal stem cells (BMSCs) are unclear. OBJECTIVE: To compare the duration, efficiency and the deviation of exogenous gene expression after rabbit BMSCs transfection using lentiviral vector and adenoviral vector which are used to mediate enhanced green fluorescent proteins (EGFP) and BMP-2. METHODS: Rabbit BMSCs at passage 5 were exposed to Ad-EGFP-BMP-2 (group A) or Lenti-EGFP-BMP-2 (group B) with multiplicity of infection of 100, as transfection groups. And in control group (group C), the same quality of culture medium was required equivalent to the groups A and B. The expression of EGFP was observed by inverted fluorescence microscope at various time intervals. And the expression of exogenous gene BMP 2 in cells was detected and analyzed by immunohistochemical staining at 72 hours after transfection as well as by western blot at 72 hours, 1, 3 weeks after transfection. RESULTS AND CONCLUSION: The intense green fluorescence emerged under the microscope at 24-48 hours after transfection in group A, which was stronger than group B, reached the peak at 72 hours, and then decreased at 1 week until disappearance at 3 weeks. No EGFP expression was detected in group C. High expression of BMP-2 was found in group A but was dramatically downregulated after 1 week. Group B showed the high expression of EGFP/BMP-2 persisted for a longer period after transfected that even lasted for 3 weeks. Overall, the lentiviral vector and adenoviral vector can efficiently transfect rabbit BMSCs and stably express the target gene of EGFP/BMP-2. Under the same MOI, compared to the adenoviral vector, transfection of lentiviral vector to rabbit BMSCs is more effectively and expression of EGFP/BMP-2 can be persistent in a longer term.

Objective:To evaluate the application of PG-SGA,NRS 2002 and BIA in nutritional assessment and screening of patients with gynecologic cancers.Methods:118 patients were randomly selected.Nutritional status were evaluated by PG-SGA,NRS 2002 and BIA,and consistency between each tools were compared.Results:The prevalence of malnutrition or nutritional risk of patients were 64.4％ (PG-SGA),57.6％ (NRS 2002),and 33.9％ (BIA) respectively.In all patients,the consistency of PG-SGA and NRS 2002 was high (P ＜ 0.001),while there were not significantly consistent between BIA and PG-SGA,or between BIA and NRS 2002 (P ＜ 0.001).Conclusion:According to the evaluation of PG-SGA or NRS 2002 in gynecologic patients,the prevalence of malnutrition or nutritional risk is high,and these two scales are suitable for nutritional assessment and screening of gynecologic cancer patients,especially in ovarian cancer patients.In addition,BIA may be a promising tool to evaluate cervical cancer patients' nutritional status.

Objective:To study the effect of microRNA (miR)-330-3p on hepatic ischemia-reperfusion injury (IRI) in mice, meanwhile, and to determine potential molecular mechanism.Methods:Eighty male C57BL/6 mice, aged 7-8 weeks, 23-25 g, specific pathogen free, were randomly divided into 8 groups (10 mice in each group) using random number table: reperfusion 2 h group, 6 h group, 12 h group, 24 h group, sham group, miR-330-3p agomir group (preoperative injection of agonist), miR-330-3p antagomir group (preoperative injection of inhibitor) and miRNA-NC group. Except for the sham group, the hepatic IRI model were established in mice. Polymerase chain reaction (PCR), Western blot and immunohistochemistry were used to detect the expression of miR-330-3p and phosphoglycerate mutase family member 5 (PGAM5), cleave caspase-1 and GSDMD. Luciferase reporter assay was performed to investigate whether miR-330-3p directly targets PGAM5. At the same time, AML12 cells were also treated with miR-330-3p mimics/inhibitor or PGAM5 siRNA, then the expression of PGAM5, NLRP3, cleave caspase-1 and GSDMD were detected by Western blot analysis.Results:Level of miR-330-3p gradually decreased after reperfusion, however, mRNA level of PGAM5 was increased thereafter ( P<0.05) as compared with the sham group. Serum level of AST and ALT were decreased in miR-330-3p agomir group while that of were increased in miR-330-3p antagomir group as a function of time following reperfusion, and the differences were statistically significant (all P<0.05). Cleave caspase-1 expression was decreased in miR-330-3p agomir group but was increased in miR-330-5p antagomir group ( P<0.05). Luciferase reporter assay was performed to determine PGAM5 was a target gene of miR-330-3p. SiRNA-mediated knockdown of PGAM5 decreased level of GAM5 (0.24±0.09), NLRP3(0.12±0.07), cleave caspase-1 (0.15±0.07) and GSDMD (1.08±0.08) as compared with the siRNA-NC group (1.17±0.14), (0.36±0.09), (0.68±0.09), (1.36±0.08), and the differences were statistically significant (all P<0.05). Conclusion:MiR-330-3p can alleviate hepatic IRI in mice, which may be related to inhibition of PGAM5-induced pyroptosis.

Objective To study the effect of subarachnoid hemorrhage (SAH) on voltagedependent potassium channels (Kv) currents of smooth muscle cells,which is hypothesized to be different between cerebral pial arteries and penetrating arteries.Methods Smooth muscle cells from cerebral pial arteries and penetrating arteries in rats were enzymatically isolated 72 h after SAH,and patch clamp was used to test the relative cell size,resting potential and Kv currents.Results Resting potential of either pial ((45.63 ±1.18) mV) or penetrating artery ((41.55-± 1.19) mV) was shifted positively by SAH,even more significantly in latter (F =8.24,P ＜ 0.05 ; F =9.36,P ＜ 0.01) ; Resting potential of pial artery of control ((38.76 ± 1.03) mV),penetrating artery of control ((38.53 ± 0.67) mV),pial artery of SAH ((36.87 ± 1.49) mV) and penetrating artery of SAH((37.89 ± 1.24) mV) were shifted positively to the same level by 1 μmol/L 4-aminopyridine (4AP; F =3.08,P ＞0.05).Maximum Current Density (Imax) of either pial ((20.82 ±0.59) pA/pF) or penetrating artery ((15.15 ±0.37) pA/pF) was compromised by SAH,also more significantly in latter (F =6.22,P ＜ 0.05) ; Imax of pial artery of control (9.15 ± 0.16),penetrating artery of control (9.04 ± 0.36),pial artery of SAH (8.77 ± 0.26) and penetrating artery of SAH (9.12 ± 0.17) were decreased to the same level by 1 μmol/L 4AP (F =2.96,P ＞ 0.05).Conclusions SAH probably shares the similar pathway with Kv blocker (4AP) in Kv currents inhibition.Further,SAH differently inhibits smooth muscle cells Kv currents and resting potential of cerebral pial arteries and penetrating arteries,which may be related with their different sensitivity towards cerebral vasospasm following SAH.

Objective To investigate the effect of pigment epithelium-derived factor (PEDF) on the pathogenesis of preeclampsia disease,by detecting the expression of PEDF in the placentas,as well as the relationship between PEDF and the production of placental vessels.Methods A study was performed in 60 cases of pregnant women with preeclampsia in the obstetrical department of Nanfang Hospital affiliated to southern medical university from October 2011 to January 2013,in which 30 cases were patients with mild preeclampsia(mPE) and other 30 cases were those with severe preeclampsia (sPE).40 normal pregnant women who also been hospitalized and delivered were selected as control group.The expression of PEDF and micro-vessel density (MVD) in placentas were assayed by using western blot and SP immunohistochemical method,then the relationship between PEDF and MVD was analyzed.Results (1) The pathological changes of placentas:the placental weight were lightened obviously in the mild preeclampsia and severe preeclampsia groups,the reduced blood vessels and luminal stegnosis were found in chorionic villus,basement membrane of trophocytes were thickening.The hyperplasia syneytiotrophoblast were like nodosity,with focus infarction,fibrinoid necrosis,or thrombogenesis.While there was no the above mentioned pathological alteration in normal control group.(2)The levels of PEDF expression in mild and severe preeclampsia group were 0.63 ± 0.09,0.93 ± 0.07,while 0.47 ± 0.04 in control group,which in mild and sever preeclampsia groups were significantly higher than that in normal group (P ＜ 0.05).Compared to mild preeclampsia group,the expression of PEDF was significantly increased in severe preeclampsia group,there was statistical significance between the difference (P ＜ 0.05).(3) The amount of microvessel density (MVD) in mild and severe preeclampsia group were 106 ±9,93 ±8,while 136 ±9 in control group,which were significantly reduced in mild and severe preeclampsia group,compared to that in normal control group (P ＜ 0.05).And it was significantly lower in severe preeclampsia group than that in mild preeclampsia group (P ＜ 0.05).(4) The expression of PEDF was negatively correlated with the amount of MVD in mild and severe preeclampsia group (r =-0.426,P ＜ 0.05 ; and r =-0.646,P ＜ 0.05 respectively),which was also negative in control group (r =-0.589,P ＜ 0.05).Conclusion Increased PEDF expression in placentas of women with preeclampsia induce the dysfunction of the placental vascular reconstruction and the pathological alteration like ischemic and hypoxia in placentas,which may be involved in pathogenesis and pathogenic progress of preeclampsia.

Objective To research the combined prognostic utility of NT-proBNP and hs-CTNI in NSTE-ACS risk stratification and early intervention therapy .Methods A total of 245 NSTE-ACS patients were divided to 4 groups(Aa ,Ab ,Ba ,Bb groups) ac-cording to immediate admission NT-proBNP and hs-CTNI .Patients were accepted percutaneous coronary intervention (PCI) ,coro-nary artery bypass grafting and conservative treatment were taken 6 months follow-up .Results The heart failure incidence of Aa group was significantly higher than Ba group(P<0 .05) .6 patients happened left main lesion or multivessel lesion in Aa group were obviously higher than other groups .The incidence of Bb group accepted PCI was obviously higher than Aa group (P<0 .05) .Bb group patients were accepted PCI earlier than other groups ,there was no significance(P>0 .05) .In 6 months follow-up ,5 patients died and 10 patients accepted revascularization again because of severe angina and AMI .Conclusion In NSTE-ACS patient ,NT-proBNP and hs-CTNI elevation was closely related with severe coronary lesions and worse prediction ,which could undergo early in-tervention therapy .