AIM: To investigate the transfection efficiency of mouse liver with non-viral vector containing manganese superoxide dismutase (Mn-SOD) gene. METHODS: The eukaryotic expression vector, gWiz/Mn-SOD, encoding human manganese superoxide dismutase was constructed. The plasmids of gWiz/Mn-SOD were mixed with cationic lipids, followed by injection into mice via branch of superior mesenteric vein, to induce Mn-SOD over-expression in murine liver detected by RT-PCR, Western blotting, SOD activity and immunohistochemical staining. RESULTS: gWiz/Mn-SOD transfection resulted in the obvious expression of exogenous Mn-SOD mRNA and protein in hepatic tissues at 8 hours after injection, and elevated mitochondria SOD activity 8.4 times in transfected hepatocytes than that in non-transfected cells at 72 hours after injection. It was showed that nearly 70% of mouse hepatocytes was obviously Mn-SOD positive after transfection. CONCLUSION: High expression efficiency of Mn-SOD gene in mouse liver is achieved safely by injection of gWiz/Mn-SOD and cationic lipid mixture into branch of superior mesenteric vein. [

OBJECTIVETo analyze the relationship between inorganic ingredients (Ca, P and F) concentration of plaque residues in the primary teeth and caries status, sugar exposure frequency.

METHODSSubjects were 93 of 3-5 years old children living in Beijing area. According to their caries status, they were classified into two groups: Severe early childhood caries(S-ECC) group and caries free(CF) group. Plaque samples were collected from subjects respectively. Ca, P and F concentration of plaque residues was measured by atomic absorption spectroscopy, spectrophotometer and fluoride electrode respectively. The information of daily sugar exposure frequency of children was obtained from a standardized questionnaire filled by their parents. The relationship between inorganic ingredients (Ca, P and F) concentration of plaque residues and daily sugar exposure frequency was analyzed and the relationship between inorganic ingredients (Ca, P and F) concentration of plaque residues and caries status in the S-ECC group was also analyzed.

RESULTSThe mean values for Ca, P and F concentration of plaque residues were lower in the S-ECC group, but the difference was not statistically significant (P > 0.05). The daily sugar exposure frequency in the S-ECC group was higher than that in the CF group. There were no relationships between Ca, P and F concentration of plaque residues and caries status (dmft, dmfs), sugar exposure frequency in the primary dentition.

CONCLUSIONPlaque residues from CF subjects did not contain a greater pool of calcium, inorganic phosphorus, and fluoride ions than that from S-ECC subjects.

Calcium ; Carbohydrates ; Child ; Child, Preschool ; Dental Caries ; Dental Caries Susceptibility ; Dental Plaque ; Fluorides ; Humans ; Phosphates ; Tooth, Deciduous

Objective To assess the relationship between human papillomavirus (HPV) and extrageni-tal Bowen's disease. Methods Regular PCR with consensus primers for LI region as well as mix primers and nested PCR were performed to detect the DNA of a broad range of cutaneous and mucosal HPV types in tissue samples from lesions of 41 patients with extragenital Bowen's disease and from normal skin of 48 human controls. Semiquantitative PCR and tyramide-based in situ hybridization (ISH) were also conducted to determine the load and localization of HPV DNA in HPV-positive samples. Results HPV DNA was detected in lesions from 5 (12%) of the 41 patients with extragenital Bowen's disease. Of the 5 HPV-positive patients, 3 carried mucosal HPV types (HPV16 in 2 cases, HPV 33 in 1 case) with a viral load of 101 to 103 copies, 2 cutaneous HPV types (HPV27 in 1 case and HPV76 in 1 case). As ISH showed, there was a generalized expression of mucosal HPV DNA in most tumor cell nuclei but not in peritumoral normal tissue, and no expression of cutaneous HPV DNA was observed in lesions. HPV DNA was detected in 1 (2.1%) control tissue sample, which proved to be epidermodysplasia verruciformis-associated HPV23. There was no significant difference in the detection rate of HPV DNA between the patients and controls. The viral load of cutaneous HPV types amounted to 10-2 to 10-3 copies in the 2 patients, which was similar to that of HPV 23 in the normal control. Conclusion Mucosal HPV types may be closely associated with the development of extragenital Bowen's disease.

To analyze the pathogenesis, clinical symptoms, misdiagnosis and operation of endometriosis in unusual sites,32 cases of unusual site endometriosis were analyzed. 468% was correctly diagnosed before operation,and 625% was found to have a history of operation.Good prognosis can be achieved by combined treated on the basis of discussing its pathogenesis clarifying its increasing incidence rate,and attaching importance to its malignant tendency.

Objective This study evaluated whether liver ischemic preconditioning has delayed protective effect on liver ischemia-reperfusion injury and probe into the protective mechanism.Methods 36 wistar rats were randomly divided into two groups, group A:ischemic preconditioning,Group B:Control. The group A rats were subjected to liver ischemic preconditioning,While group B rats were subjected only laparotomy.After 48 hours,Liver tissue samples were abstained to detect the expression of HSP70.Occlusion the inflow vessels to the left and median lobes 45-minute then reperfuse 90-minute.Liver tissue samples were abstained to detect the expression of TNF-?.Serum ALT was detected before ischemia and after 90-minute reperfusion.Results The serum ALT level in group A is much lower than that in group B (P

AIM: To examine the effect of insulin on the proliferation of human hepatocellular carcinoma cell and its possible mechanisms. METHODS: The human hepatocellular carcinoma cell line(HepG-2) was used to study the changes in calcium ion across plasma membrane (Ca 2+ APM)under the action of insulin by the assay of atomic absorption spectrum, and in the proliferation under the action of insulin and calcium ion antagonist (isoptin). RESULTS: The influx of Ca 2+ APM and the proliferation was increased after insulin administration, but the proliferation was inhibited by isoptin. CONCLUSION: Changes in the homeostasis of calcium ion across plasma membrane was involved in the effect of insulin on the proliferation of human hepatocellular carcinoma.

Objective To explore the clinical characteristics of Pierre Robin syndrome and the experience of its diagnosis and therapy, and provide the accurate diagnosis and treatment of Pierre Robin syndrome .Methods Seventeen cases of Pierre Robin syn-drome from September 2008 to August 2013 were enrolled in this study .The clinical data were analyzed retrospectively to explore the clinical characteristics and treatment of Pierre Robin syndrome .Results Seventeen cases of Pierre Robin syndrome had typical clini-cal features, including micrognathia, cleft palate or high palatine arches, glossoptosis, and feeding difficulties.The babies were given corresponding treatment according to their clinical grading. Among those babies, three cases died , including one dying of severe pneu-monia in hospital, and the other two dying of severe pneumonia and malnutrition after giving up treatment .Among 14 survivors, 12 ba-bies were followed up , including 5 babies who achieved optimal growth and development when they were one year old , and the other 7 babies who had feeding difficulties in varying degrees after discharge , and lagged behind normal children in the growth and develop-ment.Conclusions Early diagnosis, accurate classification, and individualized treatment plan for Pierre Robin syndrome might im-prove the prognosis of children with this type of disease .

AIM:To investigate the effect of decitabine (Dacogen, DAC) on the proliferation and differentia-tion of K562 cells.METHODS:The K562 cells were treated with different concentrations of DAC .The colony formation ability of the cells was detected by the colony formation assay with semi-solid culture .The cell viability was detected with MTT assay.The morphologic features were observed under inverted microscope with Wright ’s staining.The changes of the cell cycle distribution and the expression of CD 11b and CD42b were analyzed with flow cytometry .The protein expression of CDK2, cyclin E1, P27, GATA-1 and PU.1 in the K562 cells was determined by Western blot .RESULTS:DAC signifi-cantly decreased the colony number of the cells and cell viability in a dose-dependent manner .The morphological changes of the cells displayed partial differentiation .After treated the K562 cells with DAC for 72 h, the cell proportion in S phase was obviously decreased , while the cell proportion in G 2/M phase was obviously increased in a dose-dependent manner . After treated the K562 cells with DAC for 7 d, the percentage of CD11b and CD14 positive cells was further elevated , and the protein expression of P27, GATA-1 and PU.1 was increased.However, the protein expression of CDK2 and cyclin E1 was decreased .CONCLUSION:DAC inhibits the proliferation and induces differentiation of the K 562 cells via regulation of cell cycle .

ObjectiveTo explore the clinical significance and characteristics about terminal sialyl residues linked to N-glycans in the tissue and bile glycoproteins from bile duct carcinoma. MethodsThe mRNA expression of sialyltransferases, including ST3Gal-Ⅲ and ST6Gal-Ⅰ, was detected in the tissue extracts of 35 cases with bile duct carcinoma(BDC) and 35 cases of benign biliary disease (BBD) by semiquantitative reverse transcription-polymerase chain reaction(RT-PCR). With the lectin of WGA(Triticum valgaris agglutinin), which can recognise the terminal sialyl residues in N-glycans, all the specimens of BDC and BBD were subject to lectin histochemical staining, followed by computer-aided digital analysis. Lectin dot-blotting with horseradish peroxidase-labeled WGA was performed for the bile samples from each of the 35 cases of BDC and BBD. ResultsThe mRNA levels of ST3Gal-Ⅲ and ST6Gal-Ⅰ elevated in BDC(P

Objective To investigate sedation of dexmedetomidine versus midazolam on me-chanically ventilated patients.Methods Seventy-six mechanically ventilated patients were randomly divided into midazolam group (group M)and dexmedetomidine group (group D).The loading dose and maintenance dose of midazolam was 0.06 mg/kg and 0.04-0.2 mg·kg-1 ·h-1 ,and 1 μg/kg and 0.2-0.7 μg·kg-1 ·h-1 for dexmedetomidine.The drug dose was adjusted according to Ramsay score scale.Time when Ramsay reach 2-4 score,the length of ICU stay,mechanical ventilation time,drug adverse reactions,the levels of inflammation factors were recorded and compared between two groups.Results The length of ICU stay and arousing time in group D was significantly lower than that of group M (P <0.05).The levels of TNF-α,hs-CRP in group D were lower after 24 hr treat-ment than that of group M(P <0.05).The levels of hs-CRP in group D were lower after 48 hr treat-ment than that of group M (P <0.01).Conclusion Dexmedetomidine treatment reduced the lengh of ICU stay and levels of inflammatory factors of mechanical ventilated patients.