Objective To investigate the effects of Inula Britannica on myocardial caspase-3 and cytochrome c ( cyt c) following overtraining-induced acute myocardial injury in rats.Methods Forty-eight male Wistar rats weighing 200-220 g were randomly divided into 3 groups:group control (group C,n =8) ; group exhausting swim (group E,n =24) and group Inula Britannica (group IB,n =16).The animal model of overtraining-indnced acute myocardial injury was developed by exhausting swim.The animals were forced to swim until they were exhausted.The animals sank to the bottom and no righting reflex or escape response was elicited when they were taken out of water in groups E and IB.In group IB oral Inula Britannica 25 ml/kg was given 24 h and immediately before overtraining.Blood samples were taken from inferior vena cava immediately and at 6,24 h after overtraining in group E and at 6,24 h after overtraining in group IB for determination of serum cardiac troponin I (cTnI) concentration (by ELISA).The animals were sacrificed after blood sampling and myocardial specimens were obtained for microscopic examination and determination of caspase-3 and cyt c expression (by immuno-histochemistry).Results Overtraining significantly increased serum cTnI concentration and up-regulated myocardial caspase-3 and cyt c expression in group E as compared with group C.Oral Inula Britannica significantly attenuated overtraining-induced increase in serum cTnI concentration and myocardial caspase-3 and cyt c expression in group IB as compared with group E.Conclusion Inula Britannica can reduce overtraining-induced acute myocardial injury by down-regulating caspase-3 and cyt c expression.

OBJECTIVETo investigate the effects and mechanisms of diethylhexylphthalate (DEHP) on morphology and function of progenitor Leydig cells (PLC) in rats.

METHODSTwenty pregnant SD rats were randomly divided into 4 groups ( n = 5): normal control group, DEHP low dose group , middle dose group, and high dose group, which were treated from postnatal day (PND) 1 to PND 21 of the pubs with DEHP at the doses of 0, 10, 100, 750 mg/(kg · d) in 0.5 ml of corn oil by gavage respectively. At the end of the treatment, the male pups were killed and blood samples were collected for determination of serum testosterone concentration by chemiluminescence method. The body weight, testis weight and anogenital distance (AGD) were measured. The morphology of PLC was observed by light and transmission electron microscopy. The protein expression of steroidogenic acute regulatory protein(StAR) in PLC was determined by immunohistochemistry. The mRNA expression of insulin-like growth factor-I (IGF-I) in the testis was assayed by real-time PCR.

RESULTSCompared with normal control group, the serum testosterone and AGD of male pubs from the middle and high dose groups were declined significantly (P < 0.01), the testis weight and body weight from high dose group were decreased significantly (P < 0.01), while the testis weight increased in the low dose group (P < 0.05). Under light microscope, PLC showed hyperplasia and cluster aggregation in the low dose group and focal hyperplasia in the middle and high dose group. The spermatogenic cells in seminiferous tubules showed decrease, apoptosis and unfix in the high dose group. Under transmission electron microscope, the PLC showed decreased lipid droplets, smooth endoplasmic reticulum and mitochondriae in the treated group. The mRNA expression of IGF-I increased in the low dose group, and the protein expression of StAR decreased in the middle and high dose group.

CONCLUSIONLactating exposure to DEHP may interfere with the synthesis of testosterone of PLC in male pubs, the decrease of StAR and the damage of PLC may be involved in it.

Animals ; Body Weight ; Diethylhexyl Phthalate ; adverse effects ; Female ; Germ Cells ; drug effects ; Insulin-Like Growth Factor I ; metabolism ; Lactation ; Leydig Cells ; cytology ; drug effects ; Male ; Organ Size ; Phosphoproteins ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; drug effects ; Testis ; Testosterone ; blood

Objective To research the feasibility of the prefabrication of skin flap with frozen allogeneic bone.Methods From November 2010 to July 2011,fifteen miniature pigs were randomly divided into three groups(n =5) according to different implanted positions.Rewarming,deep-frozen allogenic bone was implanted in subcutaneous tissue,subfascial compartment and muscle respectively.Emission Computed Tomography (ECT) was used in the 4th week,the 8th week and the 12th week postoperatively.Angiography and pathological examination was used in the 12th week.Results The difference of the result of ECT was statistically significant betweent the 4th week and the 8th week (P ＜ 0.05),and there was not statistically significant difference betweent the 12th week and the 8th week (P ＞ 0.05).Angiography showed that all allogeneic bone was vascularized in each group.Pathological examination showed that the vascularization and the inducing osteogenesis of the allogeneic bone which was implanted in the subfascial compartment or muscle was better than that in the subcutaneous tissue.Conclusion The deep-frozen allogenic bone can be used for the prefabrication of the bone-skin flap.

Objective To investigate children′s myelodysplastic hematopoietic cells reaction to interleukin (IL)-15.Methods CD 34 + cells in bone marrow from 18 myelodysplast syndrome(MDS) patients were purified by an immunomagnetic beads sorting system. Apoptosis of hematopoietic precursors was assayed by propidium iodine staining and flow cytometric analysis.Results On 8th cultured day,when IL-15 concentration was between 0-100 ng/ml,it could suppress apoptosis of hematopoietic cells in MDS patients in a dose-and- time dependent manner. IL-15 in study group significanthy lower than that of control group.Conclusion IL-15 may partly suppress apoptosis of hematopoietic cells in MDS patients.

Objective To study the pre-transplant serum level of anti-endothelial cell antibody(AECA)in kidney allograft recipients and its impact on the episode of acute rejection (AR) within 6 months after transplantation. Methods A total of 495 kidney allograft recipients with pre-transplant serum between December 1998 and August 2003 in our center and 40 healthy controls(negative controls)were enrolled in the study.Clinical data including AR within 6 months after transplantation were analyzed retrospectively.The serum AECA level was measured by cyto-ELISA using EA.hy926 cells as substrate,which was shown as the ratio of P (patient)/N (negative control)=(A_(petient)-A_(blank contrnal)/(A_(negative contral)-A_(blank contral).AECA was considered positive when P/N value Was greater than the average A_(negative control)value plus two times the standard deviation.Results Positive rate of AECA was 18.8%(93/495).AECA level in hemodialysis patients who had been on hemodialysis more than 12 months was 1.43±0.37,greater than those less than 12 months(1.27± 0.32,P=0.013)and those of non-dialyzed patients(1.31±0.32,P=0.029).Correlation coeffieient between AECA level and hemodialysis duration was 0.218 (P=0.018).AR incidence in AECA positive recipients was 38.7%,greater than that in AECA negative recipients (23.4%,P=0.002). Incidence of acute T cell-mediated rejection and acute antibody-mediated rejction increased significantly (P=0.035,P=0.002 respectively).Multifactor logistic regression analysis indicated that AECA positive,PRA greater than 1 0%and high CDC level were risk factors of AR with odds ratio of 2.056,1.751 and 1.764 respectively(P=0.004,0.029,0.050). Conclusions The AECA positive in pre-transplant serum indicates the elevated risk of acute allograft rejection.The AECA level increases with prolonged hemodialysis duration.

Animals ; Humans ; RNA Interference ; RNA, Antisense ; genetics ; RNA, Small Interfering ; genetics ; SARS Virus ; genetics ; physiology ; Severe Acute Respiratory Syndrome ; virology

BACKGROUND:Currently, the research about effect of non-dextran coated superparamagnetic iron oxide nanoparticles on cellproliferation and cytotoxicity is relatively much less. OBJECTIVE:To evaluate the effects of 0, 25, 50, 75, 100 mg/L non-dextran coated superparamagnetic iron oxide nanoparticles on the proliferation and cytotoxicity of rat bone marrow mesenchymal stem cels. METHODS:Culture media containing 0, 25, 50, 75, 100 mg/L non-dextran coated superparamagnetic iron oxide nanoparticles were prepared for culture of bone marrow mesenchymal stem cels. After 24 hours of culture, the cels were confirmed using Prussian blue staining, and cellcounting was detected using cellcounting kit-8. Meanwhile, lactate dehydrogenase activity in the supernatant and intracelular superoxide dismutase activity were detected. RESULTS AND CONCLUSION:Loading of non-dextran coated superparamagnetic iron oxide nanoparticles in BMSCs was confirmed by Prussian blue staining. The percentage of cels labeled with non-dextran coated superparamagnetic iron oxide nanoparticles was up to 100% when the cels were incubated with a non-dextran coated superparamagnetic iron oxide nanoparticle solution of 50 mg/L and above, but 25 mg/L was insufficient to label al of the cels. Furthermore, as the concentration of non-dextran coated superparamagnetic iron oxide nanoparticles decreased, the cellproliferation rate decreased gradualy. The 25 mg/L group had a minimum cellproliferation rate, but the 25 and 50 mg/L groups showed no statisticaly significant difference (P > 0.05). Therefore, 50 mg/L is considered as the appropriate concentration of non-dextran coated superparamagnetic iron oxide nanoparticles, under which, the labeling efficiency is higher and the cytotoxicity is lower.

Objective To investigate the causes and risk factors of postpartum hemorrhage (PPH) in hepatitis B virus (HBV) infected parturient.Methods Retrospective analysis was performed on the 1021 HBV infected parturient from Shanghai Public Health Clinical Center from July 2005 to June 2011.The comparisons were done by chi-square test.Results Among 1021 cases of HBV infected parturient,868 (85.01％) were asymptomatic and the PPH rate was 2.76％ (24/868) ;the remaining 153 cases (14.99％) were chronic active hepatitis B and the PPH rate was 16.99％(26/153).The difference between two groups was statistically significant (x2 =56.541,P＜0.01).The total incidence rate of PPH was 4.89％ (50/1021) and 17 cases (34.00％) were postpartum hemorrhage＞1000mL.The causes of PPH included uterine inertia (30/50,60.00％),abnormal placenta (11/50,22.00％),dysfunction of coagulation (5/50,10.00％) and lesion of birth canal (4/50,8.00％).The risk factors of PPH included delivery mode (x2 =6.528,P=0.038),abortion times (x2 =16.269,P=0.000),delivery times (x2 =6.990,P=0.008),ALT levels (x2=56.541,P=0.000) and HBV DNA (x2 =64.706,P=0.000).Conclusions The main causes of PPH in HBV infected parturient include uterine inertia,abnormal placenta,lesion of birth canal and dysfunction of blood coagulation.PPH is correlated with abortion times,delivery times,delivery mode,liver function and HBV DNA.The incidence of PPH in parturient with chronic active hepatitis B is higher than asymptomatic parturient.

Objective To analyze and evaluate the characteristics of enzyme kinetics of CTX-M-14 type extended-spectrum β-lactamase(ESBL) with Pro167 residue substitution. Methods By molecular cloning and PCR techniques, CTX-M-14 gene was directionally cloned into plasmid pET28a( + ) from a clinical E. coli isolate and formed an expression vector to transform competent E. coli BL21 (DE3 ). Prol67 residue substitutions of P167G, P167Q, P167S and P167T were introduced to CTX-M-14 by site-directed muta-genesis based on overlap extension PCR with the former recombinant plasmid as PCR template, respectively.The wild-type CTX-M-14, recombinant CTX-M-14 protein and its variants were expressed and purified, then their steady-state kinetic parameters (Kcat, Km and Kcat/Km ) against β-lactam antibiotics were determined.Results The kinetic parameters of wild-type and recombinant CTX-M-14 had no statistically significant differences (P>0.1). The 1/Km, Kcat and Kcat/Km values of P167S variant against ceftazidime were 16-fold, 2.87-fold and 43.6-fold higher than those of recombinant CTX-M-14, respectively, and the Kcat/Km value of P167S variant against penicillin, ampicillin, cefazolin, cefuroxime, ceftriaxone, cefotaxime decreased( < 0.05). Compared with the kinetic parameters of recombinant CTX-M-14, the kinetic parameters of P167T variant against ceftazidime had no significant change, but the Kcat values of P167Q and P167G variants decreased dramatically(P<0.01). Conclusion There was no difference between the enzyme activities of wild-type and recombinant CTX-M-14. P167S variant could not only promote the enzyme affinity of CTX-M-14 to ceftazidime but also improve the conversion rate of enzyme-substrate complex in the ceftazidime hydrolysis. The comparison of the kinetic parameters of CTX-M-14 and its variants with Pro167 residue substitution showed that the increased activity of CTX-M ESBL variants against ceftazidime could not be simply explained with the enlarged cavity in active site that may be caused by the replacement of Pro167 residue by smaller amino acids.

This study assessed the clinical application of transvaginal three-dimensional ultrasound (3D TVUS) in the diagnosis of congenital uterine malformation. A retrospective study was performed on 62 patients with congenital uterine malformation confirmed hysteroscopically and/or laparoscopically. The patients were subjected to transvaginal two-dimensional ultrasound (2D TVUS) and 3D TVUS. The accuracy rate was compared between the two methods. The accuracy rate of 3D TVUS was (98.38%, 61/62), higher than that of 2D TVUS (80.65%, 50/62). 3D TVUS coronal plane imaging could demonstrate the internal shape of the endometrial cavity and the external contour of the uterine fundus. It allowed accurate measurement on the coronary plane, and could three-dimensionally show the image of cervical tube, thereby providing information for the diagnosis of some complex uterine malformation. 3D TVUS imaging can obtain comprehensive information of the uterus malformation, and it is superior to 2D TVUS for the diagnosis of congenital uterine malformations, especially complex uterine anomaly.