Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on voltage-gated calcium channels on the cell membrane of pheochromocytoma cell 12 (PC 12 cells) induced by ischemic/hypoxia and its mechanisms.Methods PC12 cells at logarithmic phase were collected,which were challenged with hypoxia for 6 hours and reoxygenation for 12 hours to stimulate the nerve cells suffered ischemia/reperfusion pathological process in vitro.PC12 cells were divided into non-infection group,infected by lentivirus containing green fluorescent protein (GFP) without HSP70 gene lentivirus control group (GFP lentivirus control group),and infected by lentivirus containing HSP70 and GFP gene recombined lentiviral infection group (HSP70 recombined lentiviral infection group).Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot were used to determine mRNA and protein expressions of L-type voltage-gated Ca2+ channel subunits cav1.2 and cav1.3,receptor gated channel subunits NR1 and NR2,and Na+/Ca2+ exchange (NCX) in PC12 cells.Results After being challenged with hypoxia/reoxygenation,the mRNA and protein expressions of cav1.2,NR1 and NR2 in the PC12 cells were significantly lower in HSP70 recombined lentiviral infection group than those of GFP lentivirus control group and non-infection group [cav1.2 mRNA (2-△△Ct):3.13 ± 0.46 vs.5.12 ± 0.52,5.13 ± 0.66;NR1 mRNA (2-△△△Ct):1.61 ± 0.44 vs.3.23 ±0.82,3.31 ±0.78;NR2 mRNA (2-△△Ct):2.09±0.41 vs.3.91 ±0.64,3.88±0.62;cav1.2 protein (gray value):2.82±0.39 vs.3.98±0.23,3.96±0.24;NR1 protein (gray value):1.84±0.35 vs.2.79±0.21,2.86±0.23;NR2 protein (gray value):0.87±0.24 vs.1.57±0.31,1.33±0.44;all P ＜ 0.01].But there were no statistical differences in the mRNA and protein expressions of cav1.2,NR1 and NR2 between GFP lentivirus control group and non-infection group (all P ＞ 0.01).There were no statistical differences in the mRNA and protein expressions of cav1.3 and NCX among non-infection group,GFP lentivirus control group and HSP70 recombined lentiviral infection group [cav1.3 mRNA (2-△△Ct):4.82 ± 0.32,4.72 ± 0.36,4.82 ± 0.29;NCX mRNA (2-△△Ct):3.49 ± 0.78,3.47 ± 0.71,3.56 ± 0.65;cav 1.3 protein (gray value):2.63±0.40,2.64±0.39,2.68±0.39;NCX protein (gray value):3.27±0.48,3.34±0.48,3.31 ±0.42;all P ＞ 0.01].Conclusion Exogenous HSP70 affects the expression of L-type voltage-gated Ca2+ channel subunit cav1.2 and receptor gated channel subunits NR1 and NR2,which may protect PC12 cells from the injury caused by ischemic/hypoxia.

Objective To investigate the clinical factors influencing the treatment efficacy of microsurgery on patients with cerebral arteriovenous malformations (AVMs). Methods One hundred and eleven patients,admitted to our hospitals from January 1,2003 to December 31,2009 and underwent microsurgical cerebral AVMs resection were chosen in our study; their clinical data were retrospectively analyzed.The functional neurological status was classified with modified Rankin scale 7 d and 6 months after surgery. Binary logistic regression models were applied to test the effect of patient gender, age,intra-cerebral hemorrhage,and the size,deep venous drainage pattern and location of AVMs on long-term and short-term treatment efficacy. Results Twenty-nine patients (26.1％) showed new neurological deficits 7 d after the surgery.Univariate analysis and multivariate analysis demonstrated that deep venous drainage, eloquent location and large size of AVMs, high Spetzler-Martin grading aneurysms and no intra-cerebral hemorrhage were significantly associated with new early neurological deficits (P＜0.05),while gender and age of the patients had no significant association with new early neurological deficits (P＞0.05).The scores of neurological status scale in patients with Spetzler-Martin grading Ⅰ,Ⅱ and Ⅲ were significantly different between 7 d and 6 months after the surgery (P＜0.05). Conclusion AVMs patients having deep venous drainage,eloquent location and large size of AVMs,and having aneurysm and intra-cerebral hemorrhage may be the risk factors of new early neurological deficit after microsurgical resection; analyzing the Spetzler-Martin grading before surgery can help to determine the treatment options.

The incidence of solitary fibroma of seminal vesicle is low, and the source of seminal vesicle is rare. A 38-year-old patient was admitted to hospital because of intermittent gross hematuria for more than one month. Seminal vesicle space occupying lesions can be seen in pelvic MRI. Laparoscopic resection of right seminal vesicle tumor was performed, and the pathological diagnosis tended to solitary fibroma. During the 5-month follow-up, the symptoms of hematuria disappeared and no recurrence.

OBJECTIVETo clone and express Streptococcus suis serotype 2 (S. suis 2) sly gene for constructing an foundation on identification of S. suis 2 protective antigen.

METHODSThe sly gene was amplified from S. suis 2 clinical isolate strain 05ZYH33 genome DNA by PCR. The gene fragment was inserted into the expression vector pET-30b(+) to build pET30b-sly. When recombinant vector pET30b-sly was identified by restriction enzyme cutting and DNA sequencing as a correct one, subsequently it was transformed to E. coli Rosetta for expression under IPTG induction. The obtained fusion protein was purified by Ni-NTA affinity chromatography. The immunologic and hemolysis activity of the purified protein was proved through Western blot and hemolysis assay respectively.

RESULTSThe PCR product was around 1500 bp. The gene segment inserted into the recombinant vector was proven to be completely identical with the sly gene sequence in the total genome sequence of S. suis 2. The target protein expressed was up to 30% of the total somatic protein under IPTG induction. The protein purity reached above 80% after purification. The protein could be recognized by human serum infected with S. suis 2 and could dissolve swine erythrocytes with the Hemolytic titer as 256.

CONCLUSIONThe expression vector pET30b-sly was successfully constructed. The target protein could be over-expressed in E. coli and possessed its biological activity after purification.

Animals ; Bacterial Proteins ; genetics ; metabolism ; pharmacology ; Blotting, Western ; Chromatography, Affinity ; Hemolysis ; drug effects ; Humans ; Recombinant Proteins ; genetics ; metabolism ; pharmacology ; Streptococcus suis ; genetics ; metabolism ; Swine

OBJECTIVETo study the current situation and factors influencing the behavior of fruit and vegetable intake in adolescents, and to discuss health education strategy related to the behavior.

METHODS1677 students were selected from primary school and middle school in urban area of Hangzhou by a multistage sampling method. The behavior of fruit and vegetable intake was surveyed by questionnaires of the patient-centered assessment and counseling for exercise plus nutrition project (PACE+) for adolescent-scale of fruit and vegetable consumption, and analyzed by Transtheoretical Model on the servings of intake, the stage of change, process of change, decisional balance and the current status of self-efficacy.

RESULTSThe average intake of fruit and vegetable in adolescents was (3.21 +/- 1.50) servings per day ("one serving" means 100 g cooked vegetable or 100 g fruit). The behavior of consuming fruit and vegetable in most of the adolescents was in the contemplation stage, accounting for 36.91% (619/1677). The process of behavior change, decisional balance (Pros) and self-efficacy existed a positive correlation with the process of stage (r(process) = 0.38, r(decisional balance (Pros)) = 0.26, r(self-efficacy) = 0.33, t values were 16.78, 11.02 and 14.31, P < 0.05). The servings of fruit and vegetable intake existed a positive correlation with stage transition and self-efficacy (beta(stage transition) = 0.665, t = 35.07, P < 0.05; beta(self-efficacy) = 0.050, t = 2.63, P < 0.05), and existed a negative correlation with decisional balance (Cons) (beta(decisional balance (Cons)) = -0.051, t = -2.84, P < 0.05).

CONCLUSIONFruit and vegetable intake of these adolescents are under the recommended dietary intake. Along with behavior stage development and self-efficacy improvement, the intake of fruit and vegetable also increased correspondingly. Intervention strategies should aim at improving the awareness of adolescents on the health benefits of fruit and vegetable intake and promoting their confidence to reach the recommended servings.

Adolescent ; Adolescent Nutritional Physiological Phenomena ; Child ; China ; Eating ; Feeding Behavior ; Female ; Fruit ; Humans ; Male ; Models, Theoretical ; Students ; Surveys and Questionnaires ; Vegetables

OBJECTIVETo study the feasibility of vector-mediated RNA interference for HER-2-positive breast cancer therapy.

METHODSA plasmid vector capable of mediating HER-2 RNA interference was constructed, and HER-2-positive breast cancer cell line SKBR-3 was transfected with this constructed vector. The expression of HER-2 mRNA and protein was analyzed by RT-PCR and Western blotting, and the growth and apoptosis of SKBR-3 cells was analyzed after transfection.

RESULTSThe expressions of HER-2 mRNA and HER-2 protein was downregulated in response to vector-mediated HER-2 RNA interference, which also resulted in tumor cell growth inhibition and increased number apoptotic cells.

CONCLUSIONHER-2 is a good target for RNA interference and RNA interference targeting HER-2 can lead to HER-2 breast cancer cell apoptosis and growth inhibition.

Apoptosis ; Blotting, Western ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Genetic Vectors ; Humans ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Receptor, ErbB-2 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Objective To investigate the expression of interleukin-3 receptor alpha(CD123)on bone marrow cells in acute myelocytic leukemia(AML)and its clinical significances.Methods By means of Fluorescence-activated cell sorer(FACS)and semi-quantity reverse transcripition polymerase chain reaction(RT-PCR),the expression of IL-3R?(CD123+)protein on CD34+CD38-cells and mRNA in BMMNCs of 62 AML patients of Tianjin Medical University General Hospital from March 2008 to January 2009 and 12 normal controls were detected respectively;Then the correlation between IL-3R? and the clinical stages of AML were analyzed.Results CD34+CD38-CD123+/CD34+CD38-and IL-3R? mRNA in BMMNC of 33 deno-vo or relapsed AML patients were higher than those of control group(P

Objective To evaluate the outcome of a health education program on sedentary behavior among primary and secondary school children through Transtheoretical Model. Methods Five primary schools and four middle schools were selected and students from grade 3 to 5 in primary schools and grade 1 to 2 in middle schools were selected to take part in the program, as control and intervention groups respectively. Corresponding intervention measures were provided to the intervention group, with phase, process and level of changing on sedentary behavior measured for both groups during the follow-up period. Comparison to the above items on the two groups was measured statistically. Results Behavior among the intervention group was gradually changed regarding: phase contemplation, preparation and maintenance, with the proportions in phase action and maintenance higher than the control group. After carrying out of intervention programs, the sedentary time on weekdays between the two groups did not show significant differences. However, the sedentary time on weekends of intervention group was (2.53 ± 1.62) hours, significantly lower than the hours from the baseline survey (2.84 ± 1.82) and the control group (2.78 ± 1.72) respectively.During the follow-up period, the average score of intervention group in the changing process was (2.98 ± 0.77), higher than the control group (2.80 ± 0.81 ). At the same time, the average score of intervention group in the process of changing, decisional balance (pros) and self-efficacy were 2.98 ±0.77, 3.06 ± 0.75 and 3.13 ± 0.72, respectively, all higher than data from the baseline survey. In the control group, the process of changing and the average score on self-efficacy had improved.Conclusion The corresponding intervention program seemed to have played a significant role among the school-aged children on their sedentary behavior during the weekend but no significant difference was found in the weekdays.

Objective To evaluate the role of polysaccharide from Phellinus igniarius (PPI) in the improvement of oxidative stress, hepatic granuloma and hepatic fibrosis in Schistosoma japonicum-iniected in mice. Methods The mouse model of schistosomiasis was established by S. japonicum cercariae infection via the abdomen. Balb/c mice were randomly assigned into 5 groups, including the healthy control group (Group A), infection control group (Group B), PPI treatment group (Group C), praziquantel treatment group (Group D) and PPI-praziquantel combination group (Group E), of 10 mice in each group. Each mouse in groups B, C, D and E was infected with (30 ± 2) S. japonicum cercariae. Then, mice in groups D and E were given praziquantel by gavage at a dose of 500 mg/kg for successive two days on day 42 post-infection, while mice in groups C and E were given PPI by gavage at a dose of 400 mg/kg for successive 30 days on day 42 post-infection. Histopathological changes of hepatic tissues were observed using hematoxylin-eosin (HE) staining, and serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN) were determined, while the activities of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), glutathione reductase (GSH-R) and glutathione (GSH) were detected in Mouse liver homogenates. The expression of transforming growth factor-beta (TGF-β) and alpha-smooth muscle actin (α-SMA) was quantified in hepatic tissues using immunohistochemistry, and the Nrf2 and Gsta4 gene expression was quantified using quantitative real-time PCR (qPCR) assay. Results Untreated mice presented typical pathological changes of schistosomal hepatic disorders, while PPI treatment effectively alleviated hepatic egg granulomas and collagen deposition. S. japonicum infection resulted in aggravation of hepatic lipid peroxidation, induction of oxidative stress, elevated serum MDA level and a reduction in the activity of GSH and antioxidant enzymes activities in mice. As compared to infected but untreated mice, PPI treatment suppressed hepatic lipid peroxidation, increased the GSH activity and restored the activity of antioxidant enzymes. In addition, PPI treatment inhibited the TGF-β signaling pathway and up-regulated the Nrf2 and Gsta4 gene expression. Conclusions PPI plays a critical role in the treatment of schistosomiasis-induced hepatic fibrosis. It may improve oxidative stress damages through up-regulating Nrf2 and Gsta4 gene expression, thereby suppressing the development of hepatic egg granulomas and hepatic fibrosis.

Objective: Analysis of the molecular characteristics of eosinophilia. Methods: Targeting sequence to 24 patients with chronic eosinophilic leukemia (CEL) with rearrangement of PDGFRA, PDGFRB, or FGFR1 and 62 patients with hyper-eosinophilic syndrome (HES). Mutation annotation and analysis of amino acid mutation using authoritative databases to speculate on possible pathogenic mutation. Results: Thirty-seven kinds of clonal variant were detected from 17 patients with CEL, no recurrent mutation site and hot spot region were found. No pathogenic mutation was detected in 19 patients with PDGFRA rearrangement, but pathogenic mutations of ASXL1, RUNX1 and NRAS were detected from 2 patients with FGFR1 rearrangement who progressed to acute myeloid leukemia and 1 patient with PDGFRB rearrangement who progressed to T lymphoblastic lymphoma, respectively. One hundred and two kinds of clonal abnormalities were detected in 49 patients with HES. The main hot spot mutation regions included: CEBPA Exon1, TET2 Exon3, ASXL1 Exon12, IDH1 Y208C, and FGFR3 L164V. CRRLF2 P224L and PDGFRB R370C point mutations were detected separately in 2 patients with HES who treated with imatinib monotherapy and achieved hematologic remission. Conclusion The pathogenesis of CEL with PDGFRA, PDGFRB or FGFR1 rearrangement is usually single, and the progression of the disease may involve other driver mutation. A variety of genes with hot mutation regions may be involved in the pathogenesis of HES, and some mutation sites are sensitive to tyrosine kinase inhibitors.