Objective To determine if fetal stem cells can enter the maternal circulation during pregnancy and re-pair the injuries of maternal heart.Methods C57 female mice at the age of 6-8 weeks were randomly assigned to three groups:sham control, surgery without pregnancy, and surgery with pregnancy ( n=8,eath group) .The control sham group was developed by opening and closing of the chest.The other two groups underwent heart surgery.The myocardial infarc-tion ( MI) model was induced by ligation of the left anterior descending coronary artery.Half of the surgical mice mated with e-GFP transgenic male mice, and another half group was not.Electrocardiogram ( ECG) and echocardiographic images were recorded at pre-operation, post-operation and postpartum.The collected data were used to evaluate the heart function. The GFP expression was detected by immunohistochemistry and q-PCR.Results When compared with the sham group, both the ischemia surgery groups with and without pregnancy, the ECG ST segment was significantly increased.This meas-urement indicated that the myocardial ischemia surgery was successful, and no significant difference in the ST segments be-tween two ischemia surgery groups was found.However, when ECG was measured in the surgical mice after postpartum, their myocardial ischemia was dramatically improved when compared with that of the ischemia surgery only mice.Echocar-diographic images also indicated that both the surgery groups had myocardial ischemia, however, no significant difference was observed in the pregnant mice before and after postpartum.The order of the cardiac function indexes from high to low was the sham group, surgery with pregnancy group, and surgery with no pregnancy group;in particular, the cardiac func-tion of pregnancy group was significantly enhanced compared with that of the surgery with no pregnancy group (P<0.05). More importantly, both immunofluorescence and q-PCR results showed that the embryonic stem cell translocation through circulation system with GFP expression in the heart of pregnancy group, while negative in other two groups.Conclusions Embryonic stem cells can be transferred into the maternal circulation of pregnant mice, and play a role in the repairing of their cardiac injuries.

Objective To explore the role of white matter injuries in the schizophrenia induced by the NMDA re-ceptor antagonist. Methods Adult male C57BL/6J mice (8 week old) were equally divided into four groups. One group was sub-chronically treated with saline solution, and the other three groups were intraperitoneally treated with MK-801 at dose of 0.025 mg/mL (M1), 0.050 mg/mL (M2) and 0.100 mg/mL (M3) in a volume 10 ml per kilogram body weight. All ani-mals were tested using Morris water maze at the 9th-15th day and using the Hole Board exploration as well as Rota Rod performance tests on the 16th day. The myelin basic protein (MBP) and the ultrastructure of the myelin sheaths in the cor-pus callosum were then examined using immunohistochemical methods, transmission electron microscope technique and stereological methods. Results The repeated sub-chronic MK-801 treatment did not induce impairment of spatial learning and memory in Morris water maze. The MK-801 treatment at 0.25 mg/kg and 1.00 mg/kg but not at 0.50 mg/kg resulted in less exploration to a new environment. The myelin staining with anti-MBP antibody was less intense in all three schizo-phrenic groups when compared to saline control group (P<0.01). Furthermore, MK-801 treatment caused pathological al-terations of the myelin sheaths including segmental demyelination of myelinated fibers and splitting of myelin sheath lamel- lae in schizophrenic groups. The ratio of the injured myelinated nerve fibers in the corpus callosum of MK-801 treated mice [M3 group, (22.42 ± 4.24)%] was significantly higher when compared to the control mice [(3.84 ± 1.35)%,P<0.01)]. Conclusions The present study demonstrated the white matter damages, mainly low MBP expression and segmental demye-lization in the corpus callosum in the mice sub-chronic treated with MK-801, indicating that the white matter changes might be involved in the schizophrenia induced by NMDA antagonist.

Objective To construct two DNA vaccines encoding Gag-Env fusion protein and Tat-Rev-Integrase(C-half)-Vif-Nef fusion protein derived from the first HIV-1 CRF01_AE isolate(AE2f) in Chi-na and to evaluate the immunogenicity in mice. Methods Two DNA vaccines were constructed by inserting the codon optimized and synthesized gag-env fusion gene and tat-rev-integrase(c-half)-vif-nef fusion gene de-rived from AE2f into mammalian expression vector pDRVISV1. 0, the generated DNA vaccines were desig-nated as pSVAE/GE and pSVAE/TRIVN, respectively, and their in vitro expression were determined by Western blot with transfected 293T cells. Mice were i. m. immunized with either pDRVI1.0 as mock control, pSVAE/GE or pSYAE/TRIVN for 4 times at two-week interval. Two weeks following the final im-munization, cellular responses to pool of HIV-1 Env, Gag, Tat, Rev, Intergrase, Vif and Nef peptides were evaluated by ELISPOT assay. Results The construction of DNA vaccine pSVAE/GE and pSVAE/TRIVN was validated by restriction enzyme digestion and bidirectional sequencing. Western blot showed a specific band at molecular mass 220×10~3 in lane of pSVAE/GE transfeeted 293T cell and a specific band at 95×10~3 in the lane of pSVAE/TRIVN. Both DNA vaccines mounted significant specific T cell responses with (3010 ± 566) SFC/10~6 splenocytes for DNA vaccine pSV AE/GE and (948 ± 737) SFC/10~6 spleno-cytes for DNA vaccine pSVAE/TRIVN, whereas the mock control of pDRVISV1.0 only raised marginal T cell responses. Conclusion Both pSVAE/GE and pSVAE/TRIVN were capable of expressing the inserted fusion immunogen genes and able to elicit vigorous cellular immune responses, therefore, these DNA vac-cines are highly immunogenic.

OBJECTIVE: To investigate the expression and significance of miRNA-324-3p and its target gene WNT2B in tissue specimens of nasopharyngeal carcinoma (NPC) specimens. METHOD: qRT-PCR was used to detect the expression of miRNA-324-3p and WNT2B mRNA, and Western blot was applied to assay the expression of WNT2B protein in 39 cases of NPC specimens and 21 cases of non-carcinoma epithelium. The relationship between their expression levels and clinicopathological characteristics and their correlation with clinical pathological parameters was analyzed. RESULT: The expression of miRNA-324-3p was significantly down-regulated decreased but WNT2B mRNA/protein increased obviously in NPC specimens (P < 0.01). A negative correlation between miRNA-324-3p and WNT2B was spotted (P < 0.05). The expression levels of these markers were closely correlated with T stage, clinic stage and cervical lymph node metastasis (P < 0.05). CONCLUSION The loss of miRNA-324-3p and ectopic WNT2B might co-induce the initiation and progression of NPC.
Carcinoma ; Glycoproteins ; genetics ; metabolism ; Humans ; Lymphatic Metastasis ; MicroRNAs ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; Neoplasm Proteins ; metabolism ; RNA, Messenger ; metabolism ; Wnt Proteins ; genetics ; metabolism

Objective To investigate the effect of protein phosphatase 5 (PP5) on lipid metabolism in the PP5 knockout (KO) mice.Methods Male PP5 KO and wild type (WT) mice at the age of 6 weeks were used in this study. In order to study the effect of high fat diet ( HFD) feeding, the body weight was measured.The liver histology was examined by HE and oil red O staining.To further verify PP5 functions in the adipogenesis, in vitro experiment was carried out using mouse embryonic fibroblasts (MEF).Western blotting and real-time PCR were performed to quantified the expression of lipid metabolism-related genes in the liver tissues.Results Compared with the WT mice, the body weight gain was slower in the KO mice.The size of the lipid droplets was smaller and the quantity was less in the KO mouse liver tissue.In vitro study revealed that the KO mouse MEF cells showed less differentiated adipocytes with smaller lipid droplets than the WT MEF cells.This observation was further confirmed by detecting the expression of adipogenesis-related genes in the HFD liver.The markers of adipocyte differentiation, such as CD36, AP2, PPARγ2, and Glut4, were significantly decreased, while energy expenditure-related markers, such as phosphorylation of GR and expression of UCP1, were significantly increased.Conclusions Protein phosphatase 5 may play a regulatory role in the mouse lipid metabolism through regulating the de-phosphorylation of p-GR and enhancing the expression of UCP1.

Objective To study the effect of infrasound on the changes of expression of NMDAR1 in hipp- ocampal cells.Methods Eighty-eight male Sprague-Dawley rats were randomized into eleven groups:control group,90 dB/1 d,7 d,14 d,21 d and 28 d infrasound exposed groups;130 dB/1 d,7 d,14 d,21 d and 28 d infra- sound exposed groups.All the animals in the test groups were put in an infrasound field with 8 Hz,90 dB or 130 dB for 2 hours daily.Immunohistochemistry methods were used to detect the changes of intracellular expression of NMDARI in hippocampal cells.Methods The expression of NMDAR1 in hippocampus after the rats were exposed to infrasound of 8 Hz,90 dB SPL showed a procedure from reducing on the 1st day to rising on the 7th and peaked on the 14th day,then to descending on 21st day and returning to the standard level on the 28th day.Exposure to infra- sound of 8 Hz,130 dB SPL induced opposite effects on the expression of NMDAR1 compared with 90 dB SPL,which showed a process of increasing,descending,reaching to the lowest,then ascending and returning to the normal.The lowest expression of NMDAR1 occurred on the 14th day in every observed hippocampal area.Conclusion 8 Hz, 90 dB/130 dB infrasound induced certain reversible reaction in the expression of NMDAR1 of hippoeampal cells in rats,which may disturb their learning and memory function.

Apoptosis ; drug effects ; Arachidonate 12-Lipoxygenase ; analysis ; physiology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Enzyme Inhibitors ; pharmacology ; Flavanones ; pharmacology ; Humans ; Lipoxygenase Inhibitors ; Multiple Myeloma ; drug therapy ; pathology

Objective This study was operated to investigate the association between urinary albumin-to-creatinine ratio (UACR) and physical situations as hypertension and prehypertension among women.Methods Blood pressure,height,weight and waist circumference were measured and factors such as cigarette smoking,alcohol intake,family history of hypertension,were investigated.Blood glucose and lipid,serum uric acid,urinary albumin and urinary creatinine were tested on 1796 women aged ≥30 years living in the Jinchang district of Suzhou.Associations between UACR and hypertension as well as prehypertension were analyzed,by using ordinal multinomial logistic regression models.Results The mean levels of UACR were 15.54 (7.67,32.53),9.01 ( 5.45,18.06),7.13 (4.60,12.50 ) mg/g and the rates of higher UACR were 27.57％,13.42％,9.61％ in hypertensive,pre-hypertensive and normotensive subjects,respectively,with significant differences noticed among the three groups (P＜0.05).The average systolic blood pressure/diastolic blood pressure appeared to be 125.3/80.9,128.8/82.7,130.8/84.0 and 135.1/85.9 mm Hg for participants with UACR in the first,second,third and fourth quartile,respectively.The risks of prehypertension or hypertension increased with increasing UACR levels.Dose-response relationship was seen between UACR and risks of prehypertension or hypertension.Multivariable adjusted odds ratios (95％CI) of prehypertension or hypertension in the upper quartiles of UACR were 1.32 ( 1.02,1.70),1.72 ( 1.32,2.24),and 2.37 (1.80,3.11 ),respectively,when compared with the lowest quartile.Conclusion Elevated UACR was associated with both hypertension and prehypertension among women.

This study was aimed to observe the effects of siRNA on Livin expression and function in K562 cells. Livin siRNA were designed and synthesized, then were transfected into K562 cells by using AMAXA nucle transfactor. Expressions of Livin mRNA and protein in transfected K562 cells was detected by RT-PCR and Western blot respectively. Non-transfected cells were used as control. The enhanced green fluorescent protein plasmid was used as positive control and the transfection efficiency was detected by flow cytometry. Cell apoptosis was measured by flow cytometry with Annexin V-FITC/PI double staining. The results showed that the transfection efficiency of electroporation method was about 50. The synthesized siRNA inhibited livin expression at both mRNA and protein levels. The rate of K562 cell apoptosis increased from (9.63 ± 0.89) in control group to (12.07 ± 1.39) and (27.41 ± 2.30) at 24 h and 48 h after transfection, respectively (P < 0.05). It is concluded that the siRNA can inhibit anti-apoptosis of livin gene via down-regulating livin gene expression, which may provide the new method for anti-leukemia study.
Adaptor Proteins, Signal Transducing ; genetics ; Apoptosis ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; K562 Cells ; Neoplasm Proteins ; genetics ; RNA, Small Interfering ; genetics

BACKGROUND5α-Reductase inhibitors (5α-RI) act by inhibiting the conversion of testosterone to dihydrotestosterone (DHT), thereby preventing DHT induced benign prostatic hyperplasia. The existing 5α-RIs can be classified into two types: competitive and noncompetitive. Currently, limited evidence is available concerning the effect differences between the two types of 5α-RI on androgens. The purpose of this study was to assess the effects of competitive and noncompetitive 5α-RIs on serum and intra-prostatic androgens in beagle dogs.

METHODSTwenty beagles with spontaneous benign prostatic hyperplasia were randomly allocated into two groups: epristeride group (n = 10) in which beagles were treated with epristeride at 1 mg/kg once a day for 3 months, and finasteride group (n = 10) in which beagles were treated with finasteride at 1 mg/kg once a day for 3 months. The levels of intra-prostatic testosterone and DHT were measured before treatment and on day one after three months medication. Serum levels of testosterone and DHT were measured at the same time points. Changes in androgen levels before and after treatment were analyzed, and comparisons were made within each treatment group and between treatment groups.

RESULTSAfter 3-month treatment, serum and intra-prostatic DHT levels all decreased significantly in both the epristeride and finasteride groups. The change of DHT in serum was significantly higher in the finasteride group (-14% and -43% in epristeride and finasteride groups respectively, with P < 0.001); however there was no significant difference in the changes of intra-prostatic DHT between the two groups (-47% and -51% in epristeride and finasteride groups, respectively, P = 0.304). The decreases in DHT levels were accompanied by reciprocal increases in serum and intra-prostatic testosterone levels. Changes of testosterone were significantly higher in finasteride group both in serum (20% and 42% in epristeride and finasteride groups, respectively, P < 0.001) and in prostate tissue (18% and 29% in epristeride and finasteride groups, respectively, P = 0.004).

CONCLUSIONSTwo types of 5α-RI have similar effects in reducing DHT in prostate tissue in beagles. Competitive 5α-RI may reduce serum DHT to a greater scale, and significantly increase testosterone in beagle serum and prostate.

5-alpha Reductase Inhibitors ; pharmacology ; Androgens ; blood ; metabolism ; Androstadienes ; pharmacology ; Animals ; Dihydrotestosterone ; blood ; metabolism ; Dogs ; Finasteride ; pharmacology ; Male ; Prostate ; drug effects ; metabolism