Objective Sphingosine kinase 1 (SPK1) has been identified as a central mediator of ischemia preconditioning, and it has been shown to protect reactive oxygen species (ROS)-induced cardiomyocytes death. The present study aims at investigating the protective effects of adenovirus-mediated sphingosine kinase 1 gene (Ad-SPK1) transfer on ischemia-reperfusion induced cardiac injury. Methods Wistar rats were anesthetized and a left thoracotomy was performed. About 5?108 PFU of Ad-SPK1 in total volume of 100?l was injected intramyocardially into four separate sites of the left ventricular wall with a 30-gauge needle. The control rats received the same injection of adenovirus carrying green fluorescent protein gene (Ad-GFP). Three days later, hearts were isolated and subjected to ischemia/reperfusion (I/R) (30min/30min) ex vivo. Heart performance was evaluated by measuring the left ventricular systolic pressure (LVSP) and left ventricular end-diastolic pressure (LVEDP). The incidence of arrhythmia was recorded. Cardiomyocyte viability was detected by the detection of the release of creatine kinase (CK). The cardiac SPK1 activity was measured using an enzyme method. The forced and the total SKP1 expression was analyzed by immunoblotting with anti-FLAG and anti-SPK1 antibodies. Results The cardiac SPK1 activity was increased by about five-fold by injection of Ad-SPK1, compared to Ad-GFP control group. A more potent performance and a lower incidence of arrhythmia were observed in Ad-SPK1-injected hearts during the reperfusion period, compared with Ad-GFP-injected ones. Enzymatic activity assay showed that creatine kinase release was also less in Ad-SPK1-injected hearts. Conclusion Adenovirus-mediated SPK1 gene transfer efficiently protects heart from injury induced by ischemia-reperfusion.

Objective To investigate the imaging characters of abdominal cocoon.Methods Six cases of abdominal cocoon proved by surgery and pathologic findings were retrospectively analyzed. Abdominal plain X-ray and CT were performed in 6 cases.The gastrointestinal barium meal series were undergone in 4 cases.The imaging findings were analyzed.Results Abdominal plain X-ray suggested intestinal obstruction in 3 of 6 cases.The gastrointestinal barium meal showed"cauliflower sign"or "concertina pattern"in all of the 4 cases;CT images revealed a conglomeration of multiple small bowel loops in all 6 cases and the intestinal loops seemed to be encapsulated in a membranelike sac.Conclusion The imaging features of gastrointestinal barium meal and CT scan could suggest the diagnose of abdominal cocoon.

Objective To investigate the effect of Zidan decoction containing serum on endometrial receptivity in uterine immunity environment,and then to discuss if it's effect on endometrial receptivity through its controlling the expression of cell factors which related to endometrial receptivity.Methods Mice's endometrium tissue samples and uterine Natural Killer (UNK) cells were obtained from pregnant mice on gd 4.5 days and gd 12.5 days.After endometrial glandular epithelial cells,endometrial stromal cells and UNK cells were separated through primary culture,epithelial cells and endometrial stromal cells at rate of 1 ∶ 1 a certain concentration UNK were mixed together and highly purified-cultured.Different concentrations of Zidan decoction was acted on the culture solution.FBS,black and aspirin control group were setup for contrast.The expression of LIF and VEGF which influenced by Zidan decoction in the immunity environment was assayed by Realtime-PCR.Results The expression of LIF and VEGF was significantly increased in the immunity environment of uterine (in 10％ Zidan decoction group,10％ aspirin group and 10％ blank control group:LIF was 2.10±0.20,1.98±0.14,0.90±0.05 separately,and VEGF was 3.17±0.31,2.52±0.09,0.92±0.06respectively,P＜0.05).Conclusion Zidan decoction could improve endometrial receptivity by increasing the expression of cell factor which related to endometrial receptivity with the help ofUNK paracrine.

Objective To explore the effects of transcranial magnetic stimulation (TMS) on motor function in patients with Parkinson's disease (PD) using meta-analysis. Methods Eight comparative studies of the effects of TMS were meta-analyzed. Results The combined studies confirmed a significant difference before and after TMS treatment. Between the experimental and control groups the effect was also highly significant. Conclusion TMS may play an active role in the rehabilitation of motor function for patients with Parkinson's disease.

Objective:To study the anti-proliferation effects of a heat shock protein 90(Hsp90) inhibitor,17-allylamino-17-demethoxygeldanamycin(17-AAG),on a human breast cancer cell line,SKBr3,and related mechanism.Methods:MTT assay was used to detect the growth inhibition of SKBr3 cells.Cell cycle and apoptosis were analyzed by flow cytometry.Alteration of human epidermal growth factor receptor 2(HER2) in SKBr3 cells being treated with 17-AAG were measured by immunohistochemistry.Results:17-AAG significantly inhibited growth of SKBr3 cells in vitro in a dose-dependent manner with an IC_(50) value at 3.09 ?g/ml.Under concentrations of 0,0.625,1.250,2.500,5.000 and 10.000(?g/ml,)the percentages of cell apoptosis were(1.03?0.08)%,(3.68?0.67)%,(7.06?1.12)%,(11.23?1.36)%,(20.32?1.98)%,and(31.65?2.96)%;the percentages of cells at G_(0)/G_(l) phase were 58.61%,54.34%,49.55%,43.73%,35.52%,and 27.46%;the percentages of cells at S phase were 29.57%,25.21%,19.65%,22.98%,19.71%,and 15.46%;the percentages of cells at G_(2) /M phase were 11.82%,20.45%,30.18%,33.29%,44.77%,and 57.08%,respectively.The level of HER2 expression in SKBr3 cells being treated with 17-AAG,compared to that in control cells,was reduced significantly.Conclusion:17-AAG can inhibit the growth of human breast cancer cell and enhance its apoptosis.It may be a promising anti-tumor drug.

Objective To investigate the protective effects of imipramine on alveolar epithelial barrier functiou in mice against LPS-induced acute lung injury (ALI),and explore the possible mechanisms.Methods Total of 32 SPF male Balb/c mice were randomly (random number) divided into four groups:control group,Imipramine group,LPS group,LPS + Imipramine group.To establish an animal model of ALI,mice were administered intraperitoneally with LPS in 20 mg/kg.Mice were treated with imipramine in 25 mg/kg 30 min prior to LPS administration.FITC-FD4 was administered in mice via the tail vein with FITC-FD4 10 min before mice sacrificed under anesthesia at 12 hours after LPS administration,then bronchoalveolar lavage fluid (BALF) and lung tissue were obtained.HE staining was used to observe histopathological changes,and pathology scores;lung tissue wet-to-dry weight ratio and BALF/serum FD4 ratio were used to assess pulmonary edema and alveolar epithelial permeability.Real-time PCR,western blot and immunochemistry were employed to detect the mRNA expressions and protein levels of Occludin,Claudin-4 and ZO-1.Data were analyzed with SPSS 16.0 software,one way analysis of variance (ANOVA) was used to compare multiple sets of variables,the intergroup comparisons were analyzed by the least-significant-difference (LSD) tests with P ＜ 0.05 for the statistically significant difference.Results Compared with LPS group,LPS + lmipramine group had a statistically significant decrease in pathological score [(9.22 ± 0.21) vs.(11.23 ± O.55),P ＜ O.05);the wet-to-dry weight ratio in LPS + Imipramine group was less than that in LPS group and the difference was statistically significant (P ＜ 0.05);compared with LPS group,the ratio of BALF/serum FD4 in LPS + Imipramine was less and the difference was statistically significant (P ＜ 0.05);compared with LPS group,the mRNA expressions and protein levels of Occludin,Claudin-4 and ZO-1 in LPS + Imipramine group were significantly increased (mRNA:Occludin:P ＜ 0.05;Claudin-4:P ＜ 0.05;ZO-1:P ＜ 0.05 . western blot:Occludin:P ＜ 0.05;Claudin-4:P ＜ 0.05;ZO-1:P ＜ 0.05).Immunochemistry showed that Occludin and Claudin-4 were present mainly in alveolar epithelial cell membrane,Z0-1 was found mainly in cytoplasm of alveolar epithelial cell.In control group and Imipramine group,tight junction proteins were obviously expressed.Compared with control group,protein levels in LPS group were significantly decreased (Occludin:P ＜ 0.05;Claudin-4:P ＜ 0.05;ZO-1:t =6.59,P ＜ 0.05);compared with LPS group,the tight junction proteins in LPS + Imipramine group were significantly increased (Occludin:P ＜ 0.05;Claudin-4:P ＜ 0.05;ZO-1:P ＜ 0.05).Conclusion The protective effects of imipramine on alveolar epithelial barrier function by up-regulating tight junction proteins expression in murine LPS-induced ALI.

Objective To summarize the clinical characteristics,diagnosis and treatment of pulmonary sequestration(PS) in children.Methods The clinical data of 19 children with PS confirmed by operation and(or) imaging examination from Mar.2003 to May.2008 were analyzed retrospectively.Thirteen cases were male,and 6 cases were female.One case was newborn infant.Seventeen cases received operation.Intralober type had a pulmonary lobectomy and extralobar type had a sequestrectomy.Chest X-ray and CT scan examinations were performed on the patients before operation.Results Seventeen cases got complete cure by operation,and the mean age at operation was 5.2 years.Chest enhanced CT indicated abnormal feeding arteries.Lobectomy was performed in 13 cases of intralobar PS,4 cases of extralobar PS were resected in the separated lung tissue,and all patients had unilateral lesions.No late deaths occurred in this group except 1 case who was complicated with malformation,and the postoperative follow-up showed an excellent recovery.Conclusions The main diagnostic methods of PS are CT and angiography.The diagnosis of PS can be confirmed when systemic feeding arteries are indicated on enhanced CT scans.Surgical resection is the main choice of treatment in all cases of PS in order to prevent recurrent infection and hemoptysis.The excellent results can be obtained by surgery.

OBJECTIVETo study the mechanism of reversal effect of Curcuma Wenyujin n-Butyl alcohol extract (CWNAE) on multiple drugs resistance (MDR) of SGC7901/VCR cells.

METHODSSGC7901/VCR cells were co-culured with different concentrations CWNAE (80, 40, and 20 μg/mL) and Verapamil (VP, 10 μg/mL) for 24 h, and then acted with Adriamycin (ADM) for 1, 2, and 4 h, respec- tively. SGC7901/VCR cells with no intervention were taken as the vehicle control group. SGC7901/VCR cells treated with ADM alone were taken as the control group. The effect of CWNAE on intracellular ADM concentration was detected by flow cytometry (FCM). Cells were treated as mentioned before without any intervention of ADM. SGC7901/VCR with no ADM intervention were taken as the control group. The effect of CWNAE on the expression of P-glycoprotein (P-gp), lung resistance protein (LRP), and glu- cosylceramide synthase (GCS) was studied by Western blot. The effect of CWNAE on the location and expression quantity of P-gp was further illustrated by immunohistochemistry (IHC).

RESULTSCompared with the ADM group, the expression ratio obviously increased in the W80, W40, W20, and VP10 groups with statistical difference (all P < 0.05). The comparative expression quantity of P-gp, GCS, and LRP in SGC7901/VCR cells was obviously higher than that of non-MDR with statistical difference (all P < 0.05). The expression quantity of P-gp and GCS could be obviously down-regulated by 80 and 40 μg/mL CWN- AE, and 10 μg/mL VP, with no effect on the expression of LRP. Results of IHC proved that P-gp was mainly expressed on the cytomembrane or in the plasma, and it was also expressed on the nuclear membrane. P-gp in different locations could all be down-regulated by CWNAE.

CONCLUSIONSCWNAE could reverse the MDR of SGC7901/VCR cell line probably by inhibiting the expression of P-gp and GCS. CWNAE had no effect on LRP that also highly expressed on SGC7901/VCR. So we supposed that CWNAE could become a potential drug to reverse MDR of highly expressed P-gp and GCS.

ATP-Binding Cassette, Sub-Family B, Member 1 ; Cell Line, Tumor ; Curcuma ; Doxorubicin ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Stomach Neoplasms

Ultrasonic microbubbles were used to open blood-brain barriers (BBB) with a reversed and limited behavior feature in the study, which could improve the brain-targeted delivery of anti-tumor drugs. The glioma rat model was prepared. Low-frequency ultrasound was combined with microbubbles to affect the permeability of BBB compared with the permeability of independently administered Evans blue (EB) crossing BBB. Time point and length of ultrasound were investigated whether they affect the permeability of BBB and the damage of brain tissue. The effect of the growth time of glioma on BBB permeability was explored. Only glioma had a very little impact on BBB permeability. However, ultrasonic microbubbles opened the BBB with the features of temporary, limited and reversed behavior and improved EB and magnetic resonance imaging contrast agent penetrating BBB. A length of 30 s ultrasound is appropriate for opening BBB and no damage of brain tissue. Drugs should be injected before ultrasound so that they enter into brain as BBB opening. Ultrasonic microbubbles can open BBB effectively and safely, which improve drugs penetrating BBB under proper time point and length.
Animals ; Blood-Brain Barrier ; Contrast Media ; Drug Delivery Systems ; Glioma ; drug therapy ; Magnetic Resonance Imaging ; Microbubbles ; Permeability ; Rats ; Ultrasonics

Objective To describe the clinical characteristics of a patient with Gitelman syndrome,and to identify the associated SLC12A3 gene mutations.Methods A suspected case of teenager-onset Gitelman syndrome was observed in our hospital.It was further confirmed by clinical manifestations and auxiliary examination.In addition,direct sequencing for the exons of SLC12A3 gene and CLCNKB gene region was conducted to identify the probable disease-associated mutations.Results The case showed characteristics of hypokalemia,hypomagnesemia,and low level of urinary calcium and onset by age of 18.By excluding the possibilities of long-term use of thiazide diuretics,laxatives,chronic vomiting and diarrhea,he was finally diagnosed as a case of Gitelman syndrome.Furthermore,by Sanger direct sequencing,2 coding variations were identified in SLC12A3 gene region,including T304M and L488P.L488P was a new heterozygous mutation.Conclusion Detection of SLC12A3 gene mutation could facilitate the diagnosis of Gitelman syndrome and improve prognosis.