Objective To observe the changes of expression and activity of heme oxygenase-1(HO-1) in lung tissue of preterm rats exposed to hyperoxia,and explore the role of HO-1 in hyperexia-induced lung injury of preterm rats.Methods Three-day-old preterm rats of standard SD were randomly assigned to hyperoxia group and air group.At the third and 7th day of exposure,the expressions of HO-1 mRNA were detected by means of reverse transcription polymerase chain reaction and the cellular distribution and expressions of HO-1 protein in the lung were measured by immunohistochemical techniques,respectivesly,and the active of HO-1 was determined also.Results On the third day,in air group,the expressions of HO-1 mRNA(0.17 ?0.08),HO-1(7.23?4.63)were mildly expressed and the activity of HO-1 was(4.32?1.57) nmol/(mg?h);compared with those of air group,the expression of HO-1 mRNA in hyperoxia group(0.72?0.33) was significantly increased(Pa

It is often necessary to construct more than one recombinant plasmids when investigating the characteristics, physchemical features and functional mechanisms of genes or proteins. Repeated sub-cloning procedures including design of primers, enzyme digestion, ligation and verification of recombinant plasmids, have to be involved with. For this reason, it has become a tendency to developing new genetic vectors which can be used in multitude of experiments. Therefore, by using pIRES vector as a backbone, here we reported the construction of a mammalian expression vector: pCMV-Myc-IRES-EGFP which contains the N-terminal c-Myc epitope tag and the enhanced green fluorescent protein (EGFP) translated in an IRES-dependent manner. This novel vector can be used to testify the efficiency of cell transfection, to collect successfully transfected cell population via cytometry, to conduct transcription and translation in vitro, to purify target proteins or to trap their interactional proteins. The availability of this vector can facilitate function study of genes.
Apoptosis Regulatory Proteins ; genetics ; metabolism ; Base Sequence ; Blotting, Western ; Cell Line ; Cloning, Molecular ; Gene Expression ; Genes, myc ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Molecular Sequence Data ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

OBJECTIVETo study the epidemiological characteristics of liver diseases in a rural population in Southern Guangxi, China.

METHODSThe enzyme immunoassays was used to detect of HBsAg and AFP. AFP positive serum samples were further examined for concentration of AFP by using a radio immunoassays. Liver morphological changes were measured with ultrasonography of type B.

RESULTSThe positive rates of HBsAg in the studied population was 17.8% (2800/15,701). The prevalence rates of viral hepatitis B, cirrhosis, primary liver cancer, clonorchiasis, fatty liver disease, alcoholic liver disease were 1.1% (173/15,701), 0.4% (63/15,701), 299.3 per 100,000 (47/15,701), 6.6% (1036/15,701), 4.8% (754/15,701) and 0.3% (47/15,701), respectively. The positive rates of HBsAg and the prevalence rates of viral hepatitis B, cirrhosis, primary liver cancer, clonorchiasis, fatty liver disease in male were significantly higher as compared with those in female (5.98 < or = chi(2) < or = 394.78, P < 0.01). No difference was observed in the prevalence rates of liver cavernous hemangioma and hepatic cysts between male and female. The prevalence rates of intrahepatic bile duct stones was significantly higher in female than in male (chi(2) = 30.80, P < 0.01). The positive rates of HBsAg and the prevalence rates of viral hepatitis B and clonorchiasis were decreased with age. But the prevalence rates of cirrhosis, primary liver cancer, fatty liver disease, alcoholic liver disease, liver cavernous hemangioma, hepatic cysts and intrahepatic bile duct stones were increased with age.

CONCLUSIONThe rural areas in the southern Guangxi are high prevalence regions of liver illness, and the male resident are even at high risk.

Adult ; China ; epidemiology ; Cross-Sectional Studies ; Fatty Liver ; epidemiology ; Female ; Hepatitis B ; epidemiology ; Humans ; Liver Cirrhosis ; epidemiology ; Liver Diseases ; epidemiology ; Liver Neoplasms ; epidemiology ; Male ; Middle Aged ; Prevalence ; Rural Population

Objective To construct luciferase reporter vector containing full-length high-mobility group box 1 ( HMGB1, GenBank NM-010439) promoter for the screening of medicine. Methods The full-length HMGB1 promoter was amplified by polymerase chain reaction ( PCR) , and then was inserted into GV238 vector to construct plasmid GV238-HMGB1-P-Luc. GV238-HMGB1-P-Luc combined with internal reference plasmid pRL was co-transfected into Hela cells ( GV238-HMGB1-P-Luc group, which served as positive control group) . Plasmid pGL3-basic combined with pRL was co-transfected into Hela cells (pGL3-basic group, which served as negative control group) . Additionally, lipopolysaccharides ( LPS, 0.2 μg/mL) was used as the activator for the positive control group (LPS group), and then sodium butyrate (SB, 10 mmol/L) was used as the inhibitor for LPS group ( SB group) . At the end of experiment hour 24, luciferase activity was detected. Results The results of digestion, amplification, sequencing and identification showed that the full length of HMGB1 promoter was 2 140 bp, and the DNA sequence was correct, without mutation. Luciferase activity in GV238-HMGB1-P-Luc group was increased as compared with that of the pGL3-basic group ( P<0.05) . Luciferase activity in the LPS group was increased ( P<0.01, compared with that of GV238-HMGB1-P-Luc group) , and then was decreased after the administration of SB ( P<0.01, compared with that of the LPS group) . Conclusion A model of luciferase reporter vector containing HMGB1 promoter has been successfully constructed. Its activity can be increased by LPS, and then is in hibited by SB. The model can be used for further screening of medicine with the activities of regulating HMGB1 promoter.

OBJECTIVETo investigate the nasal carriage of antibiotic-resistant pneumococci in children of < 5 years old in the following four cities, Beijing, Shanghai, Guangzhou and Xi'an.

METHODSA total of 647 pneumococci strains were isolated and detected. Minimal inhibition concentrations (MICs) of antibiotics were determined by E-test. Disk diffusion test was used for the measurement of antimicrobial susceptibility.

RESULTSPrevalence of penicillin non-susceptible Streptococcus pneumoniae in the four cities was 41%, with Guangzhou (60.8%) ranking first, followed by Xi'an (45%), Shanghai (37%) and Beijing (25.9%). The majority of penicillin non-susceptibility isolates (23.9% - 53.8%) had a low level of resistance (MIC 0.64 - 1.5 microg/ml). The most sensitive antimicrobials in terms of percentage of susceptible organisms were amoxicillin-clavulanic acid (99.4%), followed by ceftriaxone (92.1%); cefurxime and cefaclor were slightly more sensitive than penicillin with susceptibility of 74.8% and 77.9%. Erythromycin, tetracycline and TMP-SMZ were highly resistant (83.6%, 82.1% and 76.2% respectively). Among erythromycin resistant isolates, 100% were resistant to azithromycin, 98.6% to clarithromycin, 97.2% to roxithromycin and spiramycin, and 96.6% to clindamycin. 97.2% (141/145) were typical of the macrolides-lincosamides-streptogramons B (MLSB) resistance phenotype, and 2.8% (4/145) were M phenotype. The group of PRSP was with significantly higher rates of non-susceptibility for ceftriaxone (18.4%), cefurxime (58.6%), cefaclor (53.4%), compared with the group of PEN-S (0.5%, 1.8% and 0.2%, respectively) and the rate of multi-drug resistance in the isolates of PRSP group (92.9%) was significantly higher than that of PEN-S group (59.2%).

CONCLUSIONThe rates of penicillin and multi-drug resistance among isolates of pneumococci carried nasally in are high children and the high prevalence of multi-drug resistance in the Chinese population may be becoming one of the most serious problems in this century.

Anti-Bacterial Agents ; pharmacology ; Child, Preschool ; China ; epidemiology ; Drug Resistance, Bacterial ; Drug Resistance, Multiple ; Humans ; Infant ; Infant, Newborn ; Prevalence ; Respiratory Tract Infections ; microbiology ; Streptococcus pneumoniae ; drug effects ; isolation & purification

OBJECTIVETo study the effects of curcumin on the expression of the vascular endothelial growth factor (VEGF) and androgen-independent prostate cancer cell line PC-3, and to explore its anticarcinogenic mechanism.

METHODSPC-3 cells were treated with curcumin at the concentration of 0, 6.25, 12.5, 25 and 50 micromol/L respectively. Then the cell activity was assayed by dyed rate of Typan blue and MTT at 12, 24, 36, 48, 72 and 96 hours, the cell cycle and morphological changes observed by flow cytometry (FCM) and electronic microscopy at 24 hours, the VEGF mRNA expression measured by semi-quantitative RT-PCR, and the secreting protein levels of VEGF in the supernatants determined by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe growth of PC-3 cells was suppressed obviously by curcumin in a dose- and time-dependent manner in vitro. There were significant differences in inhibition rate among different concentration and time groups (P < 0.01). Furthermore, curcumin arrested the cell cycle of PC-3 cells in the G2/M phase in a dose-dependent manner (P < 0.01). The percentages of apoptotic cells were significantly higher in different concentration groups than in the controls (P < 0.01). Apoptosis-associated morphological changes were observed in PC-3 cells at 24 hours, and a marked decline in the expression of VEGF was noted after the exposure to different concentrations of curcumin within 24 hours.

CONCLUSIONCurcumin can suppress the growth of PC-3 cells, promote their apoptosis and arrest their cell cycle in the G2/M phase, and reduce the expression of VEGF mRNA and proteins, which may sever to explain its inhibitory effect on tumor and angiogenesis.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Curcumin ; pharmacology ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Male ; Microscopy, Electron, Transmission ; Prostatic Neoplasms ; genetics ; pathology ; ultrastructure ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

Biosimilars are biological medicinal products that are highly similar to an already licensed reference product in terms of quality, safety, and efficacy. BAT1706 is being developed by Bio-Thera Solutions, Ltd. as a proposed biosimilar candidate to bevacizumab reference product (Avastin^®). Bevacizumab acts by specifically binding to vascular endothelial growth factor A (VEGF-A), and preventing the interaction of VEGF-A with its receptors on the surface of endothelial cells, then blocking the downstream signaling pathway mediated by ligand-receptor, and inhibiting endothelial angiogenesis, thus inhibiting tumor growth. Comprehensive analytical characterization studies incorporating orthogonal analytical techniques were performed to compare the in vitro functional activities of BAT1706 and Avastin^®. BAT1706 and Avastin^® showed highly similar binding activity to multiple VEGF-A isoforms and equivalent VEGF-A neutralizing activity, as well as inhibitory activity of VEGF receptor (VEGFR)-2 tyrosine kinase autophosphorylation. Both products exhibited similar binding of the Fcγ receptors and a lack of Fc-related effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). Overall, the results demonstrate that BAT1706 and Avastin^® are highly similar in terms of in vitro functional activities.

Objective To evaluate the clinical efficacy of erythromycin and azithromycin sequential therapy with single azithromycin treatment on pediatric mycoplasma pneumonia .Methods One hundred and fourteen cases of pediatric mycoplasma pneumonia were randomly divided into two groups, with 57 cases in each group.Patients in the treatment group were treated with erythromycin combined with azithromycin , and those in the control group were treated with azithromycin , and the treatment lasted for 3 weeks.The data of clinical efficacy , clinical symptoms and hospitaliza-tion time of two groups were compared after treatment . Results The total effective rate of the treatment group was 96.49%, significantly higher than that of control group ( 77.19%) ( P<0.01 ) . The time points that fever , cough and lung rales disappeared and hospi-talization time of the treatment group were significantly lower than those of control group ( P<0.01 ).The rate of adverse reactions in the treatment group was 7.02% and 3.51% in control group but without statistical difference ( P>0.05).Conclusion Erythromycin combined with azithro-mycin treatment for pediatric mycoplasma pneumonia is notably effective .

Objective To investigate the effects of advanced oxidative protein products(AOPP)on the proliferation,apoptosis and matrix matelloproteinase-1(MMP-1)synthesis of the human gingival flbroblast(HGF).To explore the possible mechanism of the periodontal destruction acceleration in diabetes through AOPP-mediated oxidative stress.Methotis HGF were isolated bv both tissue explant cultivation technique and enzyme digestion method.The culture media with 5、50、100 mg/L AOPP-HAS were added into each experimental group,but the culture media in the control group didn't contain AOPP-HAS.MTT colorimetric assay and ELISA were used to measure the changes of HGF proliferation and the levels of MMP-1 protein from HGF at different time periods.respectively.Seventy-two hours after co-culture with 50 mg/L AOPP-HSA,cell apoptosis was detected by flow cytometry with Annexin V/PI staining.Results Compared to the control group,the growth inhibition rate of HGF in 5、50、100 mg/L AOPP-HSA group was significantly different(P＜0.05).The peak value appeared at 48 hours of co-culture[(19.01±6.28)%,(30.48±5.75)%,(39.75±4.60)%.respectively].There was a dose-dependent relationship between the growth inhibition rate and AOPP-HSA.No significant difference was detected on the apoptotic level between experimental group and the control(P＞0.05).The MMP-1 synthesis in 0.5、5、50、100 mg/LAOPP-HAS group[(55.61±1.06),(65.78±4.04),(79.24±3.09),(89.76±28.88)mg/L,respectively]was significantly higher than that in the control[(34.90±3.15)mg/L]after 72 hours coculture(P＜0.05).There was a dose-dependent relationship between MMP-1 and AOPP-HSA.Conclusions AOPP may inhibit the proliferation of HGF and such effect was not achieved through apeptosis.AOPP may increase collagen degradation by promoting MMP-1 synthesis and thus may accelerate periodontal destruction process in diabetes.

BACKGROUNDSome larger scale, randomized studies have demonstrated the superiority of drug-eluting stents (DES) over bare metal stents (BMS) for the treatment of acute myocardial infarction (AMI). This study aimed to investigate the impact of DES, in comparison with BMS, on the 2-year clinical outcomes in patients with ST-elevation myocardial infarction (STEMI).

METHODSFrom January 2002 to December 2008, a total of 1301 consecutive STEMI patients treated with coronary stenting in Shenyang Northern Hospital were prospectively registered. Patients received BMS (n = 868) or DES (n = 435) implantation in the infarction related artery according to physician's discretion. A propensity score analysis was performed and two well matched subgroups were selected (BMS, n = 288; DES, n = 288) to evaluate the 2-year clinical outcomes. The primary outcome was the occurrence of major adverse cardiac events (MACE), which was defined as a composite of all-cause death, myocardial infarction (MI), or target vessel revascularization (TVR).

RESULTSSurvival salvage analysis showed that 2-year cumulative hazards were not significantly different between the two groups with respect to TVR (2.8% vs. 3.1%, log-rank P = 0.780), stent thrombosis (1.7% vs. 4.2%, log-rank P = 0.079) and MACE (8% vs. 10.8%, log-rank P = 0.236). Multivariate analysis showed that DES was an independent protective factor of MI (HR: 0.211, 95%CI: 0.049 to 0.908) and stent thrombosis (HR: 0.327, 95%CI: 0.107 to 0.994).

CONCLUSIONDES was associated with similar 2-year clinical outcomes to those of BMS for the treatment of STEMI in daily practice.

Aged ; Angioplasty, Balloon, Coronary ; methods ; Drug-Eluting Stents ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; mortality ; therapy ; Prospective Studies ; Stents ; adverse effects ; Thrombosis ; etiology ; Treatment Outcome