This study examined the role of PI3K/Akt pathway in radiosensitization of DNA damage of cervical carcinoma cells. The 50% inhibition concentration (IC(50)) of cisplatin and docetaxel in HeLa cells was detected by Mono-nuclear cell direct cytotoxicity assay (MTT) in vitro. HeLa cells were treated by cisplatin/docetaxel of 10 percent of IC(20) alone or combined with LY294002 for 24 h, and then radiated by different doses of X-ray. The cell survival ratio was obtained by means of clone formation. One-hit multi-target model was fitted to the cell survival curve to calculate dose quasithreshold (Dq), mean lethal dose (D(0)), 2Gy survival fraction (SF(2)) and sensitization enhancement ratio (SER). The pAkt and total Akt expression was detected by Western blotting and DNA damage by neutro-comet electrophoresis. The HeLa cells were randomly divided into 7 groups in terms of different treatments: Control; radiation treatment (RT) group; LY294002+RT group; cisplatin+RT group; docetaxel+RT group; LY294002+cisplatin+RT group; LY294002+docetaxel+RT group. The apoptosis ratio of each group was measured by flow cytometry. The results showed that docetaxel and cisplatin significantly enhanced the phosphorylation of Akt in radiation-treated HeLa cells. The Dq, D(0) and SF2 in LY294002-contained groups were lower than those in docetaxel or cisplatin+RT group. The SER in the LY294002+docetaxel+RT group was 1.35 times that of the docetaxel+RT group, and that in the LY294002+cisplatin+RT group 1.26 times that of the cisplatin+RT group. The Comet electrophoresis showed that tail distance in the LY294002+cisplatin+RT group or LY294002+docetaxel+ RT group was longer than in the cisplatin+RT group or docetaxel+RT group. The apoptosis ratio in the LY294002+cisplatin+RT group or LY294002+docetaxel +RT group was higher than in the cisplatin+RT group or docetaxel+RT group. It was concluded that inhibiting PI3K/Akt pathway can increase the effect of docetaxel and cisplatin on the radiosensitivity of HeLa cells and DNA damage resulted from radiation.

OBJECTIVE The aim of this study is to investigate the correlation between the spontaneous otoacoustic emission(SOAE)and transient evoked otoacoustic emission(TEOAE)in neonate. METHODS The subjects were 224 ears of 112 newborns(59 females, 53 males). The age of subjects ranged from 2 to 4 days (mean?SD: 2.68?0.74 day years). SOAE and TEOAE were measured with ILO96. RESULTS The correlation between SOAE number and TEOAE level were found (r=0.43, P

Objective To investigate the correlation between miR-21 and chemotherapy-induced premature ovarian failure (CIPOF) by measuring the serum miR-21 level in animal models and patients. Methods CIPOF animal model was established by intraperitoneal injection of cyclophosphamide. Caudal vein blood samples were collected at the first and the 15th day after the models established. Serum miR-21 level was detected by QT-PCR. The correlations between miR-21 level with estradiol (E2) level, follicle-stimulating hormone (FSH) level and ovary structure were analyzed. From Jan. 2014 to Jun. 2015, 30 patients with CIPOF and 30 normal women of child-bearing age were enrolled in the study. Serum miR-21 levels of all cases were detected and the correlations between serum miR-21 levels and other laboratory indexes [luteinizing hormone (LH), E2, FSH and prolactin (PRL)] were analyzed. Results Compared to control group, the serum miR-21 levels in CIPOF animal models and patients were obviously lower with statistical significances (t=15.467, P=0.000; t=10.197, P=0.000). Positive correlations existed in animal models between miR-21 and E2, primordial, primary, secondary and antrum follicle numbers (r=0.750, P=0.000; r=0.403, P=0.006; r=0.704, P=0.000; r=0.783, P=0.000; r=0.849, P=0.000, respectively), and a negative correlation existed between miR-21 and FSH (r=–0.801, P=0.000). A positive correlation existed in patients between miR-21 and E2 (r=0.817, P=0.000), and negative correlations existed between miR-21 and FSH (r=–0.771, P=0.000) and miR-21 and LH (r=–0.784, P=0.000). No correlation existed between miR- 21 and PRL (r=0.204, P=0.207). Conclusion The evident decrease of serum miR-21 level in CIPOF animal models and patients suggests that miR-21 may play an important role in the process of CIPOF, and the miR-21 may be a potential prevention and therapeutic option for CIPOF.

OBJECTIVETo isolate and culture human umbilical cord mesenchymal stem cells (MSCs) and explore their biological features and ultrastructure.

METHODSAfter isolating MSCs from the human umbilical cord, the proliferation, cycle, and apoptosis were observed. The cell ultrastructure was observed under transmission electron microscope. The cytokines including vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and insulin-like growth factor-1 (IGF-1) were detected using enzyme-linked immunosorbent assay.

RESULTSHuman umbilical cord MSCs had fibroblast-like morphology and increased proliferation capability. Ultrastructural analysis showed that the MSCs had active cellular metabolism and strong migration and differentiation capabilities. Meanwhile, they could secrete anti-apoptotic cytokines such as VEGF, IGF-1, and HGF.

CONCLUSIONHuman umbilical cord MSCs can secrete many anti-apoptotic cytokine and have good biological features.

Apoptosis ; Cell Cycle ; Cell Proliferation ; Cells, Cultured ; Hepatocyte Growth Factor ; metabolism ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Mesenchymal Stromal Cells ; cytology ; metabolism ; ultrastructure ; Umbilical Cord ; cytology ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the efficient transfection of green fluorescent protein gene (GFP) mediated by recombinant adenovirus vector(Ad-GFP) to rat bone marrow mesenchymal stem cells (MSCs) in vitro.

METHODSWistar rat bone marrow-derived MSCs were separated and purified in vitro by Percoll density gradient centrifugation combined with adherent cell culture followed by identification with flow cytometry. MSCs infected by Ad-GFP were observed and the transfection efficiency was assessed by fluorescence microscope. The proliferative ability of these cells was tested by CCK-8.

RESULTSThe transfection efficiency was as high as 90.0%. Expression of GFP gene of infected MSCs was stable for 1 month after infection. There was no statistically difference in proliferative ability between the infected MSCs and non-infected ones (P>0.05).

CONCLUSIONThe infected MSCs with Ad-GFP expressed GFP with high efficiency and retain the ability of proliferation as non-infected MSCs. Transgection with Ad-GFP is a highly effective method for labeling MSCs.

Adenoviridae ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; metabolism ; virology ; Cell Proliferation ; Cells, Cultured ; Female ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; virology ; Mice ; Rats ; Rats, Wistar ; Transduction, Genetic ; methods

This study examined the role of PI3K/Akt pathway in radiosensitization of DNA damage of cervical carcinoma cells.The 50% inhibition concentration(IC50)of cisplatin and docetaxel in HeLa cells was detected by Mono-nuclear cell direct cytotoxicity assay(MTT)in vitro.HeLa cells were treated by cisplatin/docetaxel of 10 percent of IC20 alone or combined with LY294002 for 24 h,and then radiated by different doses of X-ray.The cell survival ratio was obtained by means of clone formation.One-hit multi-target model was fitted to the cell survival curve to calculate dose quasithreshold (Dq),mean lethal dose(D0),2Gy survival fraction(SF2)and sensitization enhancement ratio(SER).The pAkt and total Akt expression was detected by Western blotting and DNA damage by neutro-comet electrophoresis.The HeLa cells were randomly divided into 7 groups in terms of different treatments: Control; radiation treatment(RT)group; LY294002+RT group; cisplatin+RT group; docetaxel+RT group; LY294002+cisplatin+RT group; LY294002+docetaxel+RT group.The apoptosis ratio of each group was measured by flow cytometry.The results showed that docetaxel and cisplatin significantly enhanced the phosphorylation of Akt in radiation-treated HeLa cells.The Dq,Do and SF2 in LY294002-contained groups were lower than those in docetaxel or cisplatin+RT group.The SER in the LY294002+docetaxel+RT group was 1.35 times that of the docetaxel+RT group,and that in the LY294002+cisplatin+RT group 1.26 times that of the cisplatin+RT group.The Comet electrophoresis showed that tail distance in the LY294002+cisplatin+RT group or LY294002+do-cetaxel+RT group was longer than in the cisplatin+RT group or docetaxel+RT group.The apoptosis ratio in the LY294002+cisplatin+RT group or LY294002+docetaxel+RT group was higher than in the cisplatin+RT group or docetaxel+RT group.It was concluded that inhibiting PI3K/Akt pathway can increase the effect of docetaxel and cisplatin on the radiosensitivity of HeLa cells and DNA damage resulted from radiation.

Objective: To investigate the effects of electroacupuncture (EA) on the behaviors of rat with anxiety disorder, and the expressions of hippocampal neurotransmitters including 5-hydroxytryptamine (5-HT), norepinephrine (NE) and dopamine (DA), and the expressions of hippocampal B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X (Bax).Methods: Forty-six male Wistar rats were randomly divided into a control group (n=10), a model group (n=12), an EA group (n=12), and a drug group (n=12). Except the control group, the other three groups were established into rat models of anxiety disorder using uncertain empty bottle stimulation. Rats in the EA group and the drug group received corresponding interventions for 15 consecutive days [EA group was given EA at Baihui (GV 20) and Sanyinjiao (SP 6); the drug group was given aqueous solution of alprazolam via intragastric administration]. After intervention, all four groups received open-field test (OFT) and elevated plus-maze (EPM) for behavioral evaluations. The expressions of 5-HT, NE and DA in hippocampus were determined by fluorescence spectroscopy (FS) while the expressions of Bcl-2 and Bax proteins in hippocampus were determined by Western blot (WB). Results: The OFT horizontal scores in the control group, EA group and drug group were significantly higher than that in the model group (all P<0.05), and the difference between the EA group and the drug group was statistically insignificant (P>0.05); the OFT vertical scores in the model group, EA group and drug group were significantly lower than the score in the control group (all P<0.05). The EPM percent of open-arm entries (OE％) in the control group, EA group and drug group was higher than that in the model group (P<0.05), and the differences among these three groups were statistically insignificant (P>0.05); though the percent of open-arm total time (OT％) in the EA group was lower than that in the control group (P<0.05), the difference was statistically insignificant when compared with the drug group (P>0.05), and it was significantly higher than that in the model group (P<0.05). The expression of 5-HT in the EA group was higher than that in the control group (P<0.05); the expression of 5-HT in the EA group was significantly lower than that in the model group (P<0.05); the difference between the EA group and the drug group was statistically insignificantly (P>0.05). The expression of NE in the model group was significantly higher than that in the other three groups (P<0.05), and there was no significant difference among these three groups (P>0.05). The expression of DA in the EA group was significantly higher than that in the control group and the drug group (both P<0.05), while the difference between the EA group and the model group was statistically insignificant (P>0.05). The expression of Bax in the model group was significantly higher than that in the other three groups (all P<0.05), whereas the expression of Bcl-2 in the model group was significantly lower than that in the other three groups (all P<0.05), and the differences in both Bax and Bcl-2 among the other three groups were statistically insignificant (all P>0.05). Bax/Bcl-2 in the EA group was significantly higher than that in the control group (P<0.05) and lower than that in the model group (P<0.05), and the difference was statistically insignificant when compared with the drug group (P>0.05). Conclusion: EA shows promising effects in attenuating rats' anxiety disorder, which may be achieved by the down-regulation of the expressions of 5-HT and NE in the hippocampus and/or inhibition of hippocampal neuronal apoptosis. The efficacy is comparable to that of intervention with alprazolam.

It is necessary to control the mechanical stimuli precisely in the studies of cardiac mechano-electrical feedback (MEF). In the present study a ventricular pressure-clamping system has been developed, which can be applied to isolated-perfused rabbit hearts. Controlled by a computer, this system not only can make the left ventricle follow a command defining the same pressure wave as that during a beating cycle under physiological condition, but also deliver mechanical stimuli with a proper waveform to the ventricle at a particular time phase. This system integrates multiple functions, including perfusing, pacing, recording of electrocardiogram and monophasic action potentials, and clamping and measuring of ventricular pressures in isolated-perfused hearts. Thus, it is a distinct system for investigating the phenomena and mechanisms of cardiac MEF at organ level.
Action Potentials ; Animals ; Constriction ; Electrocardiography ; Feedback ; Heart ; physiology ; In Vitro Techniques ; Rabbits ; Ventricular Pressure

OBJECTIVETo evaluate the effect of short-term intensive therapy on blood glucose control, BETA-cell function, and blood lipid levels in newly diagnosed type 2 diabetic patients.

METHODSOut-patients with newly diagnosed type 2 diabetic patients were enrolled for intensive treatment with sulfonylureas and metformin for 12 weeks, and the therapeutic effect was evaluated.

RESULTSAfter the intensive treatment, FPG, 2 hPG, and HbA1c decreased significantly (P<0.01); HOMA-IR decreased and HOMA-B increased significantly (P<0.01), and TG, CHOL, LDL decreased significantly (P<0.01) after the treatment.

CONCLUSIONShort-term intensive treatment with glimepiride combined with metformin is safe and effective in newly diagnosed type 2 diabetic patients with HbA1c>9%.

Diabetes Mellitus, Type 2 ; drug therapy ; Drug Therapy, Combination ; Female ; Humans ; Male ; Metformin ; therapeutic use ; Sulfonylurea Compounds ; therapeutic use ; Treatment Outcome

Adiponectin is an adipocyte-derived secretory protein. It was found to be associated with insulin resistance, inflammation and arteriosclerosis. To further study the biological function and expression of adiponection in vivo, adipoenctin gene knock-out and LacZ gene knock-in mouse model was constructed. Gene targeting strategy was designed to replace part of exon 2 and exon 3 of adiponectin gene with full length LacZ gene in frame with remaining upstream ATG and signal peptide sequence of exon 2. The targeting vector (Adipo-LacZ-XpPNT) was constructed and verified by restriction enzyme digestion and sequencing. CJ7 ES cells were transfected with targeting vector linearized by NotⅠ digestion, selected in the medium containing both G418 and ganciclovoir. Resistant clones were screened by PCR and further confirmed by Southern blot for correct homologous recombinants. Chimera mice were obtained by routing microinjection of homologous recombined ES cells into blastocysts. After mating, mice heterozygous and further homozygous for adiponectin knockout and LacZ gene knock-in were established. Expression of both endogenous adiponectin and exogenous LacZ gene in mouse tissues and sera were detected by RT-PCR, Northern-blot, Western blot and ELISA. The results show that adiponectin was disrupted at both mRNA and protein levels. LacZ gene is expressed exclusively in adipose tissue of mutant mice. Its expression profile is identical to endogenous adiponection. Unexpectedly, LacZ activity could not be detected in both adipose tissue and serum although LacZ protein can be detected in adipose tissue but not in serum of mutant mice. In conclusion, mice homozygous for adiponectin knockout and LacZ gene knock-in have been successfully constructed. Mutant mice display LacZ expression profile identical to endogenous adiponectin albeit neither LacZ activity nor protein can be detected in serum of mutant mice.