Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; diagnosis ; epidemiology ; virology ; Child ; China ; epidemiology ; Female ; Genes, Viral ; Genotype ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; diagnosis ; epidemiology ; virology ; Male ; Middle Aged ; Viral Load ; Young Adult

Objectives To investigate the impact of ischemic preconditioning (IPC) on dynamics of homing of endothelial progenitor cells (EPCs) after renal ischemia reperfusion injury (IR).Methods Sixty male Sprague-Dawley rats were randomly divided into three groups after right-side kidney nephrectomy:for sham-operated rats,lumbotomy without vascular clamping was performed; IR rats were clamped renal blood vessels for 40 minutes while IPC rats were pre-treated with 15 min ischemia and 10 min reperfusion.At 3,12,24 h,and 3 days after reperfusion,the pool of circulating,kidneys,lungs and spleens were harvested.The extent of renal injury was assessed by biochemical and histological examination.The dynamics of homing of EPCs was observed by flow cytometry.Results The rats in IPC group exhibited significant improvements in renal function and morphology.Compared with IR group and sham group,the number of EPCs in blood was increased in the IPC group at 12 h and 24 h after reperfusion (P ＜ 0.05).The number of EPCs in kidney was increased at all times pointin the IPC group and IR group as compared to the sham group (P ＜ 0.05.In addition,EPCs number was increased in IPC group compared with the IR group at 12 h and 24 h [(11.36±0.66)％ vs (6.37±0.69)％,(6.31±0.70)％ vs (4.40±0.60)％,all P＜ 0.05].Compared with IR group and sham group,the number of EPCs in the lung was increased in the IPC groups at 12 h after reperfusion [(2.95±0.66)％ vs (1.78±0.59)％,(1.66±0.61)％,all P ＜ 0.05].The number of EPCs in spleen was increased in the IPC group at 72 h as compared with the IR group and sham group [(0.55±0.06)％ vs (0.34±0.07)％,(0.31±0.06)％,all P ＜ 0.05].Conclusions Endogenous EPCs may home to injured kidney after IPC.EPCs can also gather in the lungs and spleen.

Objective To report the experience of the arterial switch operation(ASO) for Taussig-Bing anomaly and late outcomes.Methods From January 2001 to December 2015,57 patients were underwent arterial switch operation for Taussig-Bing anomaly in Fudan university affiliated children's hospital cardiac center,Median age and weight at operation was 63(37.5-88.5)days, 4.1(3.4-5.0)kg, respectively.29 patients with Arch anomalies(50.9%), 23 patients with unusual coronaries(40.3%),according to have arch anomaly or not and surgery time, dividing the patients into two groups, group A(have, n=29)and group B(not have, n=28), earlier experience into group 1(2001-2008, n=27), later experience into group 2(2009-2015, n=30), respectively.Results The Mortality was12.3%, the mortality of group A and group B was 13.8%, 10.7%(P>0.05),group 1 and group 2 was 22.2%, 3.3%(P<0.05) respectively, follow up was complete in 47 patients with a mean follow-up of(6.2±3.5) years , three patients lost, there was no late mortality, the actual survival at 1, 5year was 87%, 87%, respectively.Reintervention was required in 10 patients(21.3%), the aorta-PA valve diameter ratio was a risk factor for reintervention(group A P=0.02, group B P=0.04) ,and 1,2,5year free of reintervention was 95.6%, 86.6%, 77.2%, respectively.Conclusion The ASO approach can be applied to Taussig-Bing anomaly with acceptable mortality , and it is the procedure of choice at our institution.One stage to repair TBA with aortic arch abnormalities did not influence outcomes.The aorta-PA valve diameter ratio<0.5 was a risk factor for reintervention.

Objective To investigate the immunogenicity of immunodominant cytotoxic T lymphocyte epitope E749-57 of human papilloma virus (HPV) 16 oncoprotein E7 chaperoned by heat shock protein (HSP)110. Methods Mouse HSP110 gene was cloned into prokaryotic expression vector pQE-80L for the expression of HSP110 protein, which was purified using Ni-NTA column. SDS-PAGE and Western-blot were conducted to confirm the purified mHSP110 protein, which was subsequently incubated with E749-57 peptide under heat shock condition, and high-performance liquid chromatography (HPLC) was used to evaluate the binding efficiency of the recombinant protein and E749-57 peptide. Twenty mice were divided into 4 groups to be immunized with mHSP110 protein, E749-57 peptide, mHSP110-E749-57 complex and phosphate buffered saline (PBS),respectively. Two weeks after the last immunization, spleen cells were collected from the immunized mice and divided into 2 parts: one were stimulated by E749-57 peptide followed by the detection of CD8+ INF-γ+ T cells with flow cytometry; the other one were subjected to MTT analysis for the estimation of cell proliferation. The mHSP110-E749-57 complex was also used to immunize TC-1 tumor bearing mice to observe its anti-tumor effect.Results The full-length 2577 bp-sized mHSP110 gene was amplified from mouse liver cDNA and cloned into pQE-80L vector. Direct sequencing confirmed the correctness of the cloning. SDS-PAGE and Western-blot demonstrated the successful purification of mHSP110. HPLC assay showed that the purified mHSP110 protein could bind with E749-57 to form a relatively stable protein complex. The percentage of IFN-γ+ CD8+ T cells in and proliferation index of spleen cells from the complex-immunized mice were statistically higher than those from the other 3 groups of mice. Moreover, the complex could obviously inhibit the growth of TC-1 tumor in mice. Conclusion The mHSP110-E749-57 complex could enhance the generation of specific cytotoxic T lymphocytes and exert anti-tumor effects in mice.

OBJECTIVETo investigate the differential expression of different types of Smads in keloids, normal scars and normal skins and its possible clinicopathological significance.

METHODSRT-PCR and Western blot methods were used to examine the expression of Smads mRNA and proteins level in 10 cases of keloid, in 10 cases of normal scar and in 10 cases of normal skin tissues and fibroblasts. Fibroblasts of keloid, normal scar and normal skin were cultured in vitro. The expression difference were compared and analyzed by t-test, there was statistical difference when P < 0.05.

RESULTSThe mRNA and protein expression of inhibitory Smad7 were significantly down regulated in keloid compared with normal scar (P < 0.05) and normal skin (P < 0.05). However, no significant difference of the mRNA and protein expression of Smad2, 3 and the protein expression of phosphorylation of Smad2, 3 in keloid, normal scar, normal skin tissues and fibroblasts.

CONCLUSIONSThe decreased expression of Smad7 in keloid might play a significant role in the increased TGF-beta1/Smads signal transduction, which can not be terminated by autologous negative feedback cycle.

Adolescent ; Adult ; Female ; Humans ; Keloid ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Signal Transduction ; Smad Proteins ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Young Adult

Objective To explore the effect of electroacupuncture at Baihui (DU20) and Shenting (DU24) acupoints on learning and memory ability in rats with middle cerebral artery occlusion and reperfusion (MCAO/R), and the potential mechanisms. Methods A total of 90 healthy male Sprague-Dawley rats were randomly divided into sham group (n=18) and operation group (n=72). The MCAO/R model was established by suture method in the operation group. Finally, 54 qualified rats of the operation group were randomly divided into ischemia group (n=18), electroacupuncture group (n=18) and non-acupoint group (n=18). The electroacupuncture group received electroacupuncture at Baihui (DU20) and Shenting (DU24) for 14 days. The cerebral infarction volume was measured by magnetic resonance imaging (MRI). The learning and memory ability was tested by Morris water maze. The protein expression of 5-hydroxytryptamine 1A (5-HT1A) receptor was detected by immunohistochemistry. Results There was no significant difference in the cerebral infarction volume among three groups before intervention (F=1.678, P>0.05). Compared with the ischemia group and the non-acupoint group, the cerebral infarction volume signif-icantly reduced (P<0.001);the latency significantly shortened (P<0.001) and the times crossing the platform decreased (P<0.05);the expres-sion of 5-HT1A receptor decreased in the left hippocampus (P<0.05) in the electroacupuncture group after intervention. There was no signifi-cant difference in all the indices between the non-acupoint group and the ischemia group after intervention (P>0.05). Conclusion Electroacu-puncture at Baihui (DU20) and Shenting (DU24) could effectively increase the learning and memory ability of MCAO/R rats, which might relate with inhibiting the expression of 5-HT1A receptor in hippocampus.

BACKGROUNDHigh intracuff pressure can cause severe pharyngeal complications including sore throat or hoarseness after laryngeal mask airway (LMA) removal postoperatively. Though the application of minimum effective cuff inflating volume is suggested to maintain airway sealing and adequacy of ventilation for patients receiving general anesthesia with LMA at lower level of the intracuff pressure, it is currently not a standard care in most of the anesthetic departments. In this study, the minimum effective cuff inflating volume was determined for classic LMA Well Lead™ (Well Lead Medical Co., Ltd., China) and its impact on postoperative pharyngeal complications was also explored.

METHODSPatients with American Society of Anesthesiologists physical status (I-III) undergoing the short-duration urological surgery were recruited in this trial. First, the minimum effective cuff inflating volume was determined for size 4 or 5 LMA Well Lead in the study 1. Immediately following placement and confirmation of ideal LMA position, the cuff was inflated with 5, 7, 10 ml of air and up to 30 ml at 5 ml increment. The intracuff pressure, oropharyngeal leak pressure (OLP), and inspiratory peak airway pressure under positive pressure ventilation at the corresponding cuff volume as indicated above were recorded. Second, the enrolled patients were randomly allocated into minimum effective cuff inflating volume group (MC) and routine care (RC) group in the study 2. The minimum effective cuff inflating volume was applied and maintained in MC group, whereas the cuff volume was inflated with half of the maximum cuff inflating volume recommended by manufacturer in RC group throughout the surgical procedure and stay in postanesthesia care unit prior to LMA removal. The incidence of pharyngeal complications at 0, 2, 24, and 48 h after removal of LMA and other intra-operative adverse events were also documented.

RESULTSThe intracuff pressure varied with the cuff inflating volume in a positive linear correlation manner (Y = 11.68X - 42.1, r(2) = 0.9191) under the range of 5-30 ml for size 4 LMA. In similar with size 4 LMA, the data were also showed the linear relationship between the intracuff pressure and the cuff inflating volume (Y = 7.39X - 10.9, r(2) = 0.8855) for size 5 LMA. The minimal effective cuff inflating volume for size 4 or 5 LMA was 7-9 ml in combination of considering OLP needed to maintain airway sealing during intermittently positive pressure ventilation. The intracuff pressure in MC group was lower compared with RC group (63.0 ± 3.7 vs. 126.4 ± 24.0 cmH2O for size 4 LMA; 55.6 ± 2.4 vs. 138.5± 26.8 cmH2O for size 5 LMA; P < 0.0001). The incidence of pharyngeal adverse events was lower in MC group versus the RC group at 2, 24 h after LMA removal.

CONCLUSIONSThe relationship between the cuff inflating volume and the intracuff pressure for size 4 or 5 LMA Well Lead(TM) is in a linear correlation manner at the range of 5-30 ml. The minimal cuff inflating volume is adequate for satisfactory airway sealing and consequently associated with lower incidence of postoperative pharyngeal complications for LMA Well Lead.™.

Aged ; Female ; Hoarseness ; etiology ; Humans ; Laryngeal Masks ; adverse effects ; Male ; Middle Aged ; Pharyngitis ; etiology ; Pharynx ; surgery ; Postoperative Complications ; etiology

OBJECTIVETo investigate the effect of PI-3K and p38MAPK signal pathways on the cyclooxygenase-2 (COX-2) expression induced by the epidermal growth factor (EGF) in PC-3 cells.

METHODSPC-3 cell proliferation was detected by methylthiazolyl tetrazolium (MTT) assay after stimulated by EGF (0 microg/L), EGF (10 microg/L), EGF (10 microg/L) + LY294002 (20 micromol/L) and EGF (10 microg/L) + SC203580 (20 micromol/L), and so was the COX-2 expression in the PC-3 cells by RT-PCR and Western blot assay after stimulated the same way for 24 hours. ELISA was used to determine the changes of PGE2 in the culture medium.

RESULTSLY294002 and SC203580 signficantly inhibited PC-3 cell proliferation (P < 0.05), COX-2 expression and PGE2 production after EGF stimulation (P < 0.05).

CONCLUSIONEGF can stimulate PC-3 cells into proliferation and induce COX-2 mRNA and the upregulation of its protein expression, while LY294002 and SC203580 can inhibit EGF from the above effects on PC-3 cells.

Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; genetics ; metabolism ; Dinoprostone ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Epidermal Growth Factor ; pharmacology ; Gene Expression ; drug effects ; Humans ; Male ; Phosphatidylinositol 3-Kinases ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the effects of prostate cancer cell line PC-3 conditioned medium (PC- 3-CM) on the proliferation and osteogenic differentiation of human bone marrow human basalis mesenchymal stem cells (hBMSCs).

METHODShBMSCs were isolated and culture-expanded by density gradient centrifugation from normal volunteers. PC-3 cells were cultured till the time of logarithmic growth and then transferred to a fresh medium, which, after 24 hours of incubation, was collected as PC-3-CM. Passage 3 hBMSCs were cultured in the fresh medium alone (the control group) or that with 50% PC-3-CM (the experimental group), and the effect of PC-3-CM on the proliferation activity of the hBMSCs was detected by WST-8 assay. Based on the types of medium used, the hBMSCs were divided into Groups I (control), II (50% PC-3-CM), III (osteoblast inducer) and IV (osteoblast inducer containing 50% PC-3 CM). The effects of PC-3-CM on the osteoblastic differentiation of the hBMSCs were determined by ALP staining, ALP activity detection, Von Kossa staining, and calcium quantitation.

RESULTSAt 1, 3, 5 and 7 days of incubation, the absorbance values of the cells in the experimental group were 0.4370 +/- 0.0285, 0.7980 +/- 0.0213, 1.9090 +/- 0.0612 and 2.3023 +/- 0.0610, and those in the control group were 0.4060 +/- 0.0223, 0.6643 +/- 0.0075, 1.3727 +/- 0.0176 and 1.7947 +/- 0.0115, respectively, with significant differences between the two groups (P < 0.01) except on day 1 (P > 0.05). The positive rate and intensity of ALP staining were gradually increased in the four groups, with the ALP activities of 0.29 +/- 0.03, 1.30 +/- 0.03, 2.13 +/- 0.08, and 3.80 +/- 0.03, respectively (P < 0.01), and so was the intensity of Von Kossa staining, with the calcium depositions of 0.04 +/- 0.01, 0.44 +/- 0.05, 0.98 +/- 0.03, and 1.27 +/- 0.04, respectively (P < 0.01).

CONCLUSIONPC-3- CM can promote the proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells.

Bone Marrow Cells ; cytology ; drug effects ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media, Conditioned ; pharmacology ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Osteoblasts ; cytology ; drug effects ; Osteogenesis ; drug effects ; Prostatic Neoplasms