Objective To investigate the methylation status of the promoter of RASSF2A gene in the tissues of cervical cancer and its impacts in tumorigenesis.Methods Methylation-specific PCR (MSP) and RT-PCR techniques were applied to detect methylation status of RASSF2A gene promoter region and RASSF2A expression in tissues of 62 cervical cancer and 30 normal cervical specimens,respectively.Results Methylation frequency of RASSF2A gene in cancer tissue was higher than that in normal tissue (53.2 ％ vs 13.3 ％,P ＜ 0.05).The mRNA expression of RASSF2A in cancer tissue were less than those in normal tissue significantly (0.254 3 ±0.102 7 vs 0.623 4 ± 0.063 8,t =0.932,P ＜ 0.05).The mRNA expressed of RASSF2A gene was lower in methylated promoter group than that in unmethylated promoter group in cancer tissue samples significantly (0.118 2 ± 0.035 2 vs 0.301 2 ± 0.076 4,t =0.481,P ＜ 0.05).Conclusion Hypermethylation of RASSF2A gene in promoter region,which lead to the loss of RASSF2A gene expression,might play an important role in tumorigenesis of cervical cancer.

Objective To investigate the effects of external fixator with dynamic device under low frequency and controlled micromovement on the callus calcification and mechanical property of healing bone.MethodsForty-five sheep were performed transverse osteotomy with a gap of 2 mm on the mid-shafts of both tibias,and the hind limbs were fixed with unilateral external fixators connected to a controlled micromovement device.Ten days after osteotomy,one hind limb of each sheep was randomly selected for micromovement(30 min/d).According to different micromovement frequencies,the sheep were randomly divided into 3 groups: 0.5 Hz group,1 Hz group and 5 Hz group(n=15).The amplitude of micromovement was 0.25 mm and the micromovement stopped by the end of the fourth week postoperation.The other hind limb of each sheep was served as control group without micromovement.Morphometry of callus was done at the end of 4,6 and 9 weeks after osteotomy.Bone formation velocity,bone mineral density and biomechanical properties were compared at the end of 9 weeks.Results The areas of mineralized bone and osteoid in different miromovement groups were larger than that of control group at the end of 4,6 weeks postoperation(P

Streptozotocin-induced diabetic model was prepared in Wistar rats and the blood glucose levels were controlled at 3 levels of＜10,10-14 mmol/L or＞16.7 mmol/L by insulin,and changes of glucose transporter(GLUT)1 and 3 protein expressions of brain were observed in control rats and diabetic rats with different blood glucose levels by immunohistochemistry.The results showed that chronic hyperglycemia could decrease the protein expressions of GLUT 1 and GLUT3.

Humans ; Aconitine/analysis* ; Chromatography, High Pressure Liquid ; Poisoning/diagnosis*

OBJECTIVEThe present study was undertaken to design antisense oligodeoxynucleotides (AS-ODNs) of glial glutamate transporter-la (GLT-1a) and to evaluate the effectiveness of the designed AS-ODNs on the expression of GLT-1a.

METHODSFive sequences of GLT-1a AS-ODNs were designed according to the C terminus specific sequences of GLT-1a mRNA using antisense design software of IDT Com- pany. Western blot analysis was used to evaluate the inhibition effects of the five GLT-1a AS-ODNs on the expression of GLT-la.

RESULTSThe sequence of GLT-1a AS-ODNs with sequence of 5'-GGTTCTTCCTCAACACTGCA-3' could specifically inhibit the expression of GLT-1a in the hippocampal CA1 subfield of rats, while it had no effect on the expression of GLT-1b. This sequence showed similar inhibition on the expression of GLT-la in sham and ceftriaxone (Cef)-treated rats. It could also significantly inhibit the cerebral ischemic preconditioning (CIP)-induced up-regulation in the expression of GLT-1a. The magnitude of the inhibition in sham, Cef- or CIP-treated rats was similar by more than 60%.

CONCLUSIONFrom the designed five sequences of GLT-1a AS-ODNs, we obtained an effective sequence which can specifically inhibit the expression of GLT-1a.

Animals ; CA1 Region, Hippocampal ; metabolism ; Excitatory Amino Acid Transporter 2 ; antagonists & inhibitors ; metabolism ; Ischemic Preconditioning ; Oligonucleotides, Antisense ; genetics ; RNA, Messenger ; Rats ; Up-Regulation

The protoplasts of Micromonospora purpurea,the high yield strains of gentammicin producer were mutagenized by diethyl sulfate(DES)and ultraviolet radiation(UV)respectively,then fused,screened by gentamicin resistance and regenerated.The average fermentation unit 2200?U/ml could be achieved by shake flask for 10 batches.The average fermentation unit 1900?U/ml could be obtained by 5L fermentor for 7 batches.The quality of the end product conformed to CP2000,BP2000 and USP26 pharmacopoeia.

Objective To analyze our clinical results of head and neck reconstruction using microsur- gical free flap transfer techniques.Methods The free flap donor sites with long vascular pedicle and large diameter of vessel were routinely chosed,and chose receipt vessels with large diameter and proper position, and perform vessel ananstomosis under surgical loups instead of microscope.The un-buried free flap with a mo- nitoring window were harvest,and do double venous anastomoses in some flaps to ensure adequate venous out- flow.Results From May 1999 to March 2005,1066 consecutive free flap transfers were used to reconstruct head and neck defects.The overall success rate of free flap was 98.3%.The vessel thrombosis rate was 3.1%,and the flap salvage rate was 45.5%.Conclusion Head and neck reconstruetion using microsurgi- cal free flap transfer technique is safe and reliable,and good clinical results can be obtained.

OBJECTIVETo observe the effects of Xingpi Yang'er Granule (XYG) on serum gastrin (GAS), plasma motilin (MOT), and somatostatin (SS) in children patients with pneumonia induced diarrhea.

METHODSRecruited were 120 children inpatients with pneumonia induced diarrhea at the Department of Pediatrics, Liaocheng People's Hospital from June 2011 to June 2012. They were randomly assigned to two groups, the treatment group and the control group, 60 in each group. Those in the treatment group were treated with XYG, while those in the control group were treated with Live Combined Bifidobacterium and Lactobacillus Tablets. Besides, 30 healthy children who received physical examinations at our hospital were recruited as the healthy control group. The clinical efficacy, changes of GAS, MOT, and SS contents were observed.

RESULTSThe total effective rate was 95.0% in the treatment group and 93.3% in the control group, showing no statistical difference (P > 0.05). Compared with healthy control group, the GAS and MOT contents increased, and SS decreased before treatment in the other two groups (P < 0.05). Compared with the same group before treatment, GAS and MOT contents obviously decreased, and SS increased in the other two groups after treatment (P<0.05). Compared with the control group at the same time point, GAS and MOT decreased, and SS increased in the treatment group after treatment, showing statistical differences (P < 0.05).

CONCLUSIONSThe levels of GAS, MOT, and SS were obviously changed in children patients with pneumonia induced diarrhea. XYG had obvious regulation on their GAS, MOT and SS contents.

Child, Preschool ; Diarrhea ; blood ; drug therapy ; etiology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gastrins ; blood ; Humans ; Infant ; Male ; Motilin ; blood ; Phytotherapy ; Pneumonia ; complications ; Somatostatin ; blood

Objective To evaluate the pattern of energy metabolism and nutrients intake in patients with chronic viral hepatitis and posthepatitic cirrhosis to effectively direct their nutrition therapy.Methods Resting energy expenditure (REE) was measured with open-circuit indirect Jorimetry in 60 patients with chronic viral hepatitis and 60 patients with posthepatitic cirrhosis.Their normal basal energy expenditure (BEE) was predicted by Harris-Benedict equation and energy intake (EI) was determined by diet recall. Correlation between REE and indicators for nutrition assessment was analyzed.Results REE was (77? 21) kJ?kg~(-1)?d~(-1) in 60 patients with pusthepatitic cirrhosis,significantly lower than BEE[(95?16) kJ? kg~(-1)?d~(-1)(P0.05,and their EI was (127?34) kJ?kg~(-1)?d~(-1),1.41?0.43 times as REE,in which PROI was (1.02?0.29) g?kg~(-1)?d~(-1),1.31?0.61 times as PROE (0.87?0.34) g?kg~(-1)?d~(-1),also indicating a negative nitrogen balance (-2.02?4.07).REE,EI and intake of three nutrients,serum level of albumin and prealbumin (PA) and body weight significantly decreased in patients with posthepatitic cirrhosis,as compared to those in patients with chronic viral hepatitis (P

OBJECTIVETo observe the effect of Fn-TPO gene modification on human bone marrow mesenchymal stem cells (MSCs).

METHODSRetroviral vector containing Fn-TPO gene was constructed and bone marrow MSCs was modified by this vector. The transcription of Fn-TPO gene in MSCs was observed. The proliferation capacities, hematopoietic cells adhering capacities and TPO secretion capacities of gene modified MSCs were assayed respectively. Cord blood CD34 cells were seeded on the gene modified MSCs layers and several essential growth factors were added. After co-culturing in vitro for 7 days, the number of CD34 cells and their colony forming capacities were assayed by flow cytometry and semisolid culture assay.

RESULTSRetroviral vector containing Fn-TPO gene was successfully constructed and bone marrow MSCs were modified by this vector. Fn-TPO gene was expressed by bone marrow MSCs after gene modification. The viability of MSCs had no significant difference between pre- and post-gene-modification [(7.18 +/- 0.89) 10(4)/ml vs. (6.92 +/- 0.77) 10(4)/ml, P > 0.05]. The hematopoietic cells adhering ability of gene modified bone marrow MSCs was reinforced(0. 188 +/- 0.018 vs. 0.167 +/- 0.017, P < 0.01). The concentration of TPO in the MSCs culture supernatant raised from (5.58 +/- 0.37) ng/ml to (7.46 +/- 0.59) ng/ml (P < 0.01) and did not significantly decline in a short-time period, but influenced by the growth status of MSCs. After co-culturing with gene modified MSCs for 7 days, the absolute number of nucleated cells, the percentage of CD34+ cells and the colony numbers of BFU-E, CFU-GM, CFU-GEMM were (29.9 +/- 2.7) x 10(4), (33.3 +/- 2.8)% , 109.3 +/- 4.1, 163.7 +/- 7.1, 13.3 +/- 1.5, respectively, being significantly higher than that co-cultured with non-modified MSCs.

CONCLUSIONSFn-TPO gene modification can improve the capacity of human bone marrow MSCs for hematopoietic cells adhering, TPO secretion and cord blood CD34 cells amplification.

Bone Marrow Cells ; cytology ; metabolism ; Cell Adhesion ; Cell Proliferation ; Cells, Cultured ; Fibronectins ; genetics ; Gene Fusion ; Genetic Vectors ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Retroviridae ; genetics ; Thrombopoietin ; genetics ; metabolism ; Transfection