Objective The aim of this study was to investigate the possible effect of glycated low density lipoprotein ( LDL ) on the proliferation, differentiation, and expression of low-density lipoprotein receptor-related protein5(LRP5)mRNA,dickkopf-1(DKK1)mRNAinmouseosteoblasts(MC3T3-E1cells). WhethertheWnt signaling pathway was involved in the above process was explored else. Methods Mouse MC3T3-E1 cells were culturedwithvariousconcentrationsofglycatedLDL(glycatedLDLlevelwas2.4%,5.3%,8.7%,13.9%)for24, 48, 72 h. Proliferation of MC3T3-E1 cell was measured by CCK-8, the osteocalcin level in the medium was determined by ELISA and the expression of LRP5 mRNA, DKK1 mRNA was measured by realtime PCR. Results After cells being incubated with 5. 3% of glycated LDL for 24 h, the inhibition of MC3T3-E1 cells was more marked than that in control group(P<0. 01). The higher glycated LDL level, the severer the inhibition. The effect of LDL on the MC3T3-E1 cell proliferation was time- and dose-dependent under certain conditions. Osteocalcin level in cell culture fluid with glycated LDL was lowered significantly compared with control group. Meanwhile, the expression of DKK1 mRNA was increased significantly but expression of LRP5 mRNA decreased (P<0. 01) in MC3T3-E1 cells cultured with 5. 3% of glycated LDL for 24 h. Conclusions Proliferation and differentiation of the osteoblasts in mice can be inhibited significantly by glycated LDL. The possible mechanism in the role played by glycated LDL on the proliferation and differentiation in MC3T3-E1 cells seems to be related to expression of LRP5 mRNA, DKK1 mRNA in the Wnt signaling pathway.

Bioimprinting is a new developed technique to improve the characteristics of enzymes.Bioimprinting by lauric acid was conducted to improve the esterification activity of lipase PS in sol-gel immobilization process with methyltrimethoxysila(MTMS) and tetramethoxysila(TMOS) as the precursors.Results generated by checking the esterification activity and scanning electron microscope showed that bioimprinting can enhance the specific activity and thermal stability of lipase PS.The bioimprinting system was optimized by orthogonal experiment,and the optimal condition for lipase bioimprinting is water/silane molar ration(R) 12,polyethylene glycol(PEG) 120?l,and lauric acid 0.15 mmol.Compared with the free enzyme and the non-imprinted enzymes,the specific activity of imprinted enzymes has been improved 44.3 fold and 2.4 fold,respectively.Imprinted lipase show better thermal stability,and the relative activity is 58% after incubated in 80 ℃ for 0.5 h,while no activity was detected for the free enzyme.

Objective To study the effect of Jingyebaidu granules on treating cytomegalovirus ( CMV) infection during mid-pregnancy. Methods The sexually mature guinea pigs with no CMV infection history served as the subjects. Put the male and female ones in the same cages. Then the female ones were randomly divided into three groups during mid-pregnancy. Model control group:15 guinea pigs which were inoculated 1 mL suspension of GPCMV intraperitoneally. Jingyebaidu Medicine group:15 guinea pigs which were treated with Jingyebaidu(3. 09 mL·kg-1 ) through stomach perfusion after inoculation for 14 days. Normal control group:15 normal mid-pregnant guinea pigs. Viremia rates were examined 7 days after infection. All animals were sacrificed 20 days after infection. The placenta infection rate, pup infection rate, still-born rate were examined. Results Compared with the normal controls, the still-born rate was increased in model control group(8. 33% vs 34. 55%, P<0. 05). In comparison to the model control group, the GPCMV maternal infection rate(86. 67% vs 33. 33%), placenta infection rate (91. 67% vs 61. 22%), pup infection rate(90. 91% vs 48. 28%), still-born rate(34. 55% vs 15. 52%) were significantly decreased in the Jinyebaidu group (all P<0. 05). Conclusion Jinyebaidu granules could reduce maternal infection,pup loss, and placenta infection caused by CMV inoculation during mid-pregnancy.

Objective To investigate the value of three-dimensional speckle tracking imaging (3D-STI) in assessment of left ventricular (LV) strains. Methods Thirty healthy young adults examined by two-dimensional speckle tracking imaging (2D-STI) and 3D-STI. And the results of LV measurements were compared, which included mean peak systolic longitudinal strains, radial strains and circumferential strains. Also, the time consumption of these two methods was compared. Results The time needed for 3D-STI in acquisition and analysis of the images were (309.3±23.4)s, (305.5±11.2)s, while the time for 2D-STI were (490.6±14.4)s, (1261.4±39.9)s. The differences were signiifcant(t=-21.81, 69.94, both P<0.01). The global mean peak systolic radial strains was (48.59±7.68)%by 3D-STI and (33.25±7.27)%by 2D-STI. The difference was signiifcant(t=9.16, P<0.01). The global mean peak systolic longitudinal and circumferential strains were (-17.66±3.14)%, (-17.13±2.29)% by 3D-STI and (-21.35±2.46)%, (-21.97±3.84)% by 2D-STI. The differences were signiifcant(t=5.33, 5.99, both P < 0.01). The 3D-STI strains were different at different levels of LV. The longitudinal, circumferential and radial 3D-STI strains were largest at middle levels. However, 2D-STI strains didn′ t show such trend. Peak strains measured by 3D-STI and 2D-STI showed high inter-observer and intra-observer agreement in Bland-Altman chart. Conclusion 3D-STI is a novel, convenient and reproducible method to evaluate the strains of LV.

This retrospective analysis compared standard regimen of doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) with the dose-dense ABVD regimen (ABVD-21) in terms of efficacy and toxicity. Patients who had early-stage unfavorable or advanced Hodgkin's lymphoma (HL) according to German Hodgkin Study Group criteria from March 1999 to February 2011 were analyzed for treatment response, long-term survival and hematological toxicity. There were 85 patients in the ABVD-21 group and 118 patients in the ABVD group respectively. The complete remission rates after completion of treatment were 92.9% and 90.7% for ABVD-21 and ABVD, respectively. During a median follow-up period of 62 months, no significant difference was found in projected 10-year progression-free survival (PFS) and overall survival (OS) rates (84.7% and 94.1% respectively for ABVD-21; 81.4% and 91.5% for ABVD). Subgroup analyses showed that ABVD-21 was significantly better than ABVD for patients with IPS≥3 in terms of PFS and OS rates. Grade 3 to 4 leukopenia (51.8% vs. 28.8%, P=0.001) and neutropenia (57.6% vs. 39.0%, P=0.009) were more common with ABVD-21. We were led to conclude that dose-dense ABVD did not result in better tumor control and overall survival than did ABVD for early-stage unfavorable HL. However, patients at high risk, for example, with IPS≥3, may benefit from dose-dense ABVD.

Objective To investigate the pathological distribution and clinical characteristics of pineal region tumors (PRTs) with variant pathology.Methods We retrospectively analyzed the clinical and pathological data of 133 patients with PRTs that were performed surgical removal from January 2000 to January 2009 in our hospital.Results Sixty-one patients (45.9%) were diagnosed as having germ cell tumors,17 (12.8%) pineal parenchymal tumors,28 (21.1%) gliomas and 27 (20.2%)other resource tumors.Sex ratio of patients with germ cell tumors,pineal parenchymal tumors and gliomas were 14.25:1 (male:female),2.4:1 and 1.15:1,respectively; their median ages were 15.3,24.7 and 28.1,respectively.The serum immunologic test showed abnormal results in 33 patients; except for 1 with metastatic tumors,the others were diagnosed as having germ cell tumors.Conclusion PRTs have many pathological types and patients with PRTs are mainly diagnosed as having germ cell tumors.Correct diagnosis can not be made by imaging,serum immunologic test or biopsy, thus,obtaining complete clinical specimen during the total tumor removal is very necessary in studying the pathology of PRTs.

OBJECTIVETo investigate the biological effects of overexpression of the human DNA polymerase (pol-beta) on cellular response to DNA damage.

METHODSThe cell strain HLFbeta from the stable overexpression of the human pol-beta was contaminated with methyl methanesulfonate (MMS) for investigating the effects of the pol-beta on the cellular responses to DNA damage on the aspects such as the DNA damage, the cell cycle and the induced mutation rate.

RESULTSThe cell HLFbeta from the stable overexpression of the human pol-beta was obtained through the screening. The cellular response to DNA damage of HLFbeta induced by the MMS in the intermediate and high dosage group (ranging from 0.5 to 0.8 mmol/L) was significantly lower than that in the control group. The analysis for the cell cycle distribution showed that both the two types of cells contaminated by MMS had retardation at G(2) phase. In the HLFbeta group, the cells had the obvious G(2) phase retardation and 49.0% of the cells were retarded at G(1) phase as well when the MMS was increased to 0.5 mmol/L while in the control, only 20.1% of the cells were retarded at the G(1) phase when the same dosage of MMS was administered. Moreover, the MMS-induced mutagenesis in HLFbeta was increased from 4.5 x 10(-6) to 8.2 x 10(-6), significantly higher than that in the control group (P < 0.05).

CONCLUSIONHigh Pol-beta level decreases cellular DNA damage induced by MMS. Nevertheless, the overexpression of Pol-beta can also increase error-prone DNA synthesis during DNA repair process.

Cell Cycle ; drug effects ; genetics ; Cell Line ; DNA Damage ; genetics ; physiology ; DNA Mutational Analysis ; DNA Polymerase beta ; biosynthesis ; genetics ; DNA Repair ; Dose-Response Relationship, Drug ; Humans ; Methyl Methanesulfonate ; toxicity ; Mutagens ; toxicity ; Mutation

OBJECTIVETo construct pEGFP-C1-T vector, an eukaryotic expression plasmid of hMTH1 gene antisense RNA.

METHODSThe conservative region of hMTH1 gene was amplified by RT-PCR after total RNA being extracted from human embryo lung fibroblast (HLF) and then cloned into pGEM-T vector. After the recombinant plasmid was certified by DNA sequencing, the conservative region of hMTH1 gene was inserted into pEGFP-C1 vector reversedly and pEGFP-C1-T vector was constructed. The efficiency of antisense inhibition was verified by Western blotting after cell transfection.

RESULTS423 bp fragment including conservative region of hMTH1 gene was obtained by RT-PCR. After cloned by pGEM-T vector and certified by DNA sequencing, pEGFP-C1-T vector was successfully constructed by means of recombinant DNA technology. Additionally pEGFP-C1-T vector could efficiently decrease hMTH1 protein level by 46%.

CONCLUSIONThe efficient expression vector of hMTH1 gene antisense RNA, pEGFP-C1-T has been constructed successfully.

DNA Repair Enzymes ; Genetic Vectors ; genetics ; Humans ; Phosphoric Monoester Hydrolases ; genetics ; Plasmids ; RNA, Antisense ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the effect of cerebral state index (CSI) in measuring the level of sedation during target controlled infusion (TCI) of propofol in patients of different ages.

METHODSForty ASA class I-II patients undergoing general anesthesia were divided into group A (65 to 79 years old, n=20) and group B (20 to 55 years, n=20). The sedation level was assessed using OAA/S scale. Anesthesia was induced with TCI of propofol. The target effect-site concentration (CE) was set initially at 0.5 µg/ml followed by increments of 0.5 µg/ml every 5 min until 5 min after the patients lost consciousness and did not respond to pain stimulation (OAA/S=0). OAA/S score was recorded every 20 s, and MAP, HR, SPO(2) and CSI were recorded. Spearman correlation coefficient between OAA/S score and CSI and their prediction probabilities (Pk) were calculated. The values of CE(05), CE(50), CE(95) and CSI(05), CSI(50), CSI(95) at loss of verbal contact (LVC) (OAA/S=2) and loss of consciousness (LOC) (OAA/s≤1) were also calculated.

RESULTSCSI was well correlated to the sedation depth. The values of CE(50) and CSI50 were 1.3 µg/ml and 69.7 at LVC in group A, and were 1.8 µg/ml and 65.9 at LVC in group B, respectively. The values of CE(50) and CSI(50) were 1.5 µg/ml and 64.3 at LOC in group A, as compared to 2.5 µg/ml and 54.8 at LOC in group B, respectively. When the OAA/S scale was lower than 3, the CSI values in group A were significantly higher than those in group B (P<0.05).

CONCLUSIONCSI can effectively and rapidly distinguish the level of sedation in different age groups. At the same OAA/S scale, the target effect-site concentration in the elderly is obviously lower than that in the young patients, but CSI values were significantly higher in the elderly than in the young patients during TCI of propofol.

Adult ; Aged ; Consciousness ; Deep Sedation ; Electroencephalography ; Female ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; Monitoring, Intraoperative ; Propofol ; administration & dosage ; Young Adult

OBJECTIVETo construct DNA double-strand break (DSB) repair protein hKu70 deficient cell strain and to observe its biological characters for studying the functions of hKu70 gene and the effects of occupational harmfulness factors on DSB repair.

METHODSHuman lung fibroblasts (HLF) were transfected with the eukaryotic expression plasmids of hKu70 gene antisense RNA (pEGFP-C1-K) to construct hKu70 protein deficient cells (named as "HLFK"). The protein expression levels of hKu70 gene in HLFC and HLFK were detected by the Western blotting to estimate the effects of antisense inhibition. Morphology, growth character and growth status in soft agar of transfected HLFK were observed.

RESULTSpEGFP-C1-K vector was successfully expressed in HLF. The protein expression level of hKu70 gene in HLFK was decreased by 42% as compared with that in HLFC. No obvious changes of the biologic characters were observed in HLFK.

CONCLUSIONThe hKu70 protein deficient cell strain was successfully constructed. The hKu70 protein deficiency alone didn't induce obvious changes of the biological characters in HLFK.

Antigens, Nuclear ; analysis ; Cell Division ; DNA Damage ; DNA Helicases ; DNA Repair ; DNA-Binding Proteins ; analysis ; deficiency ; Humans ; Ku Autoantigen ; RNA, Antisense ; Transfection