Objective To establish a cell line with repressed expression of p53 by transfecting a plasmid construct expressing short hairpin RNA(shRNA)targeting p53 into human skin fibroblasts(HSFs),and to evaluate the effect of repression of p53 expression on the senescence in HSFs.Methods The eukaryotic expressing plasmid pGCsi-p53 containing shRNA targeting p53 gene was transfected into HSFs with lipofectamine.Subsequently,the cells were selected by G418,and resistant cell clones were chosen and expanded.Reverse transcription-PCR and real time fluorescence-based quanitative PCR were performed to determine the expression of p53 gene,and Western blot to detect the expression of p53 protein in HSFs.The senescence in HSFs was evaluated by SA β-gal staining,and cell proliferation by methyl thiazolyl tetrazolium(MTT)assay.Results A HSF clone with repressed expression of p53 was established successfully.The expressions of p53 mRNA and protein were downregulated in transfected HSFs compared with untransfected HSFs(0.09 ± 0.03 vs.0.32 ± 0.04,0.11 ± 0.04 vs.0.84 ± 0.05,both P ＜ 0.01).The percentage of senescent cells was 13.47％ ± 1.01％ in the transfected HSFs,significantly lower than that in untransfected HSFs(18.10％ ± 0.66％,P ＜ 0.05).As MTT assay showed,the proliferation was accelerated in transfected HSFs compared with untransfected HSFs(P ＜ 0.05).Conclusions The repression of p53 expression decelerates the senescence in HSFs,but promotes the proliferation of HSFs.

Rituximab is the first monoclonal antibody that has been approved for the treatment of lymphoma. Rituximab has shown significant efficacy in the treatment of B-cell non-Hodgkin's lymphomas, such as diffuse large B-cell and follicular lymphomas. Maintenance therapy with rituximab has further improved the prognosis in patients with follicular lymphoma. These patients responded to induction treatment. This antibody treatment has been recommended in treatment guidelines. The treatment strategy for lymphoma has continuously improved. Recent studies focused on how to improve the definition of the indication for maintenance therapy and how to optimize the current maintenance regimens. In this review, we summarized the main studies and the most recent advances on ritux-imab maintenance therapy in patients with lymphoma.

Objective: To observe two methods of cardioplegia under intrograde and retrograde, using mixed hypothemic blood in 59 patients receiving coronary artery bypass graft (CABG). Method:During perioperation and postoperation 24 hours,hemodynamie parameters were recorded. Myocardial enzyme level,the changes of ECG on 12 leads in postoperation,the frequency of using inotrope,time of staying in ICU and automatical reovery heart beat rate after openning aortic clamp were tested. Result: Cardioplegia by mixed hypothemie blood had better protective effects to the heart function, all patients could left the ICU safely,without significant differences between the two methods of cardioplegia,but in the retrograde group,there was obvious reduce in the change of myocardial enzyme level and less change of ECG in postoperation compared with those in the intrograde group. Conclusion:Both methods of mixed hypothemic blood for cardioplegia in CABG can protect effetively myocardium, but retrograde eardiaoplegia has better effect.

Objective To observe the changes of systemic and pulmonary hemodynamics during veno-venous bypass in liver transplantation Methods During the anhepatic phase, extracorporeal veno-venous bypass (EVVB) was utilized in 20 patients undergoing live transplantation Systemic and pulmonary hemodynamics were monitored through Swan-Ganz catheter during whole procedures Results As compared with the preoperative values , MAP remained unchanged in normal range during perioperative period;CO,CI,LVSW and RVSW decreased significantly during anhepatic phase and increased markedly 15 min after hepatic reperfusion (P

Objective To assess the lung inflammatory response to cardiopulmonary bypass (CPB). Methods Twenty ASAⅡ-Ⅲ patients of either sex (9 male, 11 female) aged (43 ? 12)yr, undergoing elective cardiac valve replacement were studied. Patients with liver or kidney dysfunction and lung inflammatory diseases were excluded. Premedication included intramuscular morphine 5mg and atropine 0.1mg. Anesthesia was induced with propofol 1.5-2.0mg? kg-1, fentanyl 5?g?kg-1 and vecuronium 0. 1mg?kg-1 and maintained with isoflurane (

ObjectiveTo observe the clinical effects of mirabilite external application and neostigmine injection at Zusanli acupoint combined with Xuebijing intravenous injection on patients with severe sepsis.Methods A history-prospective controlled study was conducted. Patients with severe sepsis admitted to the Fourth People's Hospital of Jiangsu University from January 2012 to November 2015 were enrolled. Twenty-one cases admitted from January 2014 to November 2015 were assigned as a research group and treated with application of mirabilite external application, Zusanli acupoint injection of neostigmine combined with intravenous Xuebijing injection; 22 patients with Xubijing treatment from January 2012 to November 2015 were included in a Xubijing group; 21 patients with routine therapy from January 2012 to November 2013 were included in a control group. The changes of white blood cell count (WBC), C-reactive protein (CRP), procalcitonin (PCT), mean arterial pressure (MAP), oxygenation index (OI), serum creatinine (SCr), acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) score, sequential organ failure assessment (SOFA), gastrointestinal function score, blood platelet count (PLT), plasma prothrombin time (PT), activated partial thromboplastin time (APTT), and D-dimmer in three groups before and after treatment were observed; the length of stay in ICU and the 28-day mortality were compared among the three groups. Results Compared with those before treatment, WBC, PCT, CRP, SCr, APACHE Ⅱ score, SOFA score, gastrointestinal function score, APTT, PT, and D-dimer were all obviously lower after treatment for 7 days in various groups; OI, MAP, PLT were significantly higher, and the improvement degree of WBC, PCT, CRP, SCr, APACHE Ⅱ score, SOFA score, gastrointestinal function score, OI in research group was more significant than those of control group and Xubijing group [WBC (×109/L): 7.52±0.75 vs. 12.87±4.13, 10.88±0.66, PCT (μg/L): 1.14±0.55 vs. 6.32±1.39, 3.47±1.94, CRP (mg/L): 21.0±9.2 vs. 65.0±13.6, 35.0±13.9, OI (mmHg, 1 mmHg = 0.133 kPa): 357.0±20.4 vs. 295.0±20.4, 309.0±21.4, SCr (μmol/L): 7.89±2.35 vs. 14.33±9.17, 11.27±4.65, APACHE Ⅱ score: 10.38±0.75 vs. 18.27±2.57, 13.09±4.10, SOFA score: 1.05±0.66 vs. 6.01±2.33, 3.26±1.03, gastrointestinal function score: 0.31±0.11 vs. 2.01±0.46, 1.85±0.29, all P < 0.05], the improvement of PLT, APTT, PT, D-dimer indicators in the research group was more significant than those in control group, but the above improvement compared with that in Xubijing group, no statistically significant difference was found [PLT (×109/L): 220.32±32.44 vs. 105.56±44.03, 170.56±54.03, APTT (s): 30.16±5.23 vs. 39.09±10.11, 29.56±6.85, PT (s): 10.74±1.25 vs. 13.56±4.65, 10.05±1.44, D-dimer (mg/L): 1.13±1.98 vs. 4.03±2.65, 1.13±0.54]. The length of stay in ICU was shortened in research group than that in control group and Xubijing group (days: 13.23±9.45 vs. 25.22±15.46, 18.56±10.33, P < 0.05); the 28-day mortality was significantly lower in the research group than that in the control group [28.6% (6/21) vs. 47.6% (10/21), P < 0.05], but there was no statistically significant difference compared with that in Xubijing group [28.6% (6/21) vs. 31.8% (7/22), P > 0.05]. Conclusions The treatment of mirabilite external application and neostigmine injection at Zusanli acupoint combined with Xuebijing intravenous injection can obviously improve the clinical symptoms, blood coagulation indexes and organ functions, reduce the levels of inflammatory indexes, shorten the time of the length of stay in ICU and elevate the survival rate of patients with severe sepsis.

Objective To evaluate the efficacy and advantages of the technique by combined PCNL and retrograde intrarenal surgery (RIRS) in a second stage to treat the complex renal stones in solitary kidney cases.Methods PCNL most with a single 18-24 F tract was performed first and RIRS was carried out at a second stage in solitary kidneys of 21 patients,of which congenital in 14.3％ (3 cases),contralateral nephrectomy in 42.8％ (9 cases),and functional solitry kidneys in 42.8％ (9 cases).Of the 21 patients,the average age was 45 years with 15 males and 6 females.The stones were 8 multiple,6 partial staghorn,and 7 complete staghorn with a mean size of 4.6 (3.8-6.8) cm.Results Of the 21 PCNL cases,a single tract,double,triple tracts were established in 18 (85.7％),2 (9.5％),1 (4.8％) cases,respectively.Mean operation time of PCNL was 95 (45-175) min.After 1 day of PCNL,all case had residual stones with a mean size of 1.9 (1.0-3.5) cm.Two case occurred fever after PCNL and one case presented bleeding resolved by selective renal artery embolization.The mean operation time of RIRS was 72 (35-95) min.Stone-free rate after RIRS was 85.7％ (18/21).The final stone free rate increased to 95.2％ (20/21) after one case received a second-look PCNL and two cases accepted ESWL.Two cases occurred fever and steinstrasses after RIRS resolved by rigid ureteroscopy.At the 3 months follow-up,renal function became stable,improved and worse in 71.4％ (n=15),23.8％ (n=5),and 4.8％ (n=l) of patients.Conclusions PCNL combined with RIRS could be an effective and safe option for complex stones in solitary kidneys with less bleeding,reduced tracts,minor complications and good renal function preservation.

Objective To determine the median effective doae (ED50) for motor block after intrathecal ropivacaine and bupivacaine. Methods Sixty ASA Ⅰ or Ⅱ patients, aged 18-64, weighing 46-75 kg, undergoing elective urological surgery under combined spinal-epidural anesthesia, were randomized into 2 groups ( n = 30each) receiving intrathecal 0.5% ropivacaine and 0.5% bupivacaine respectively. The ED50 was determined by up-down sequential allocation. The initial dose was 4 mg. Each time the dose increased/decreased by 1 mg. Efficacy was determined by the occurrence of any motor block in either lower extremity (modified Bromage scale > 0)within 5 or 10 min after the spinal injection. Results The intrathecal ED50 for motor block was 6.68 mg for ropivacaine (95% confidence interval 6.27-7.13 mg) and 4.07 mg for bupivacaine (95% confidence interval 3.56-4.47mg) . The relative motor blocking potency ratio was ropivacaine/bupivacaine 0.61. Conclusion The potency of intrathecal ropivacaine is lower than that of bupivacaine for motor block.

Objective To study the pathogenesis of osteoporosis in patients with liver cirrhosis following hepatitis B virus(HBV) infection(HBV-liver cirrhosis).Methods Bone mineral density(BMD) was measured by NM-300 sigle-energy X-ray acsorptiometry system for 61 patients with HBV-liver cirrhosis and 30 age-matched healthy controls.Serum levels of calcium regulatory hormones(CRH),1,25-dihydroxycholecalciferol [1,25(OH)_2D_3],parathyroid hormone(PTH),calcitonin(CT),osteocalcin(BGP),interleukin-1?(IL-1?),interleukin-6(IL-6) and tumor-necrosis factor-?(TNF-?),and urine level of crosslaps were measured in these patients and controls.Patients with HBV-liver cirrhosis were divided into two groups,osteoporotic and non-osteoporotic,based on their BMD in the ulna and radius,for comparison.Results BMD of the ulna and radius in patients with HBV-liver cirrhosis were significantly lower than those in controls.Serum levels of 1,25(OH)_2D_3 and BGP in patients with HBV-liver cirrhosis were significantly lower than those in controls,and much lower in osteoporotic group than that in non-osteoporotic one.While urine level of crosslaps increased significantly in patients with HBV-liver cirrhosis than in controls, much more increased in the those with osteoporosis than those without osteoporosis.Serum level of 1,25(OH)_2D_3 correlated with serum levels of BGP and BMD of the ulna and radius.Urine level of crosslaps correlated inversely with BMD of the ulan and radius in osteoporotic patients,but not in non-osteoporotic ones.Serum levels of IL-1?,IL-6 and(TNF-?) and urine level of crosslaps increased significantly in patients with HBV-liver cirrhosis than in controls,as well as increased significantly in those with osteoporosis than in those without osteoporosis.Serum levels of IL-1?,IL-6 and TNF-? and urine level of crosslaps correlated inversely with BMD of the ulna,particularly more significantly in those with osteoporosis than those without osteoporosis.Conclusions Two factors,less osteogenesis and more osteolysis,existed in patients with HBV-liver cirrhosis.Serum level of 1,25(OH)_2D_3 and serum levels of IL-1?,IL-6 and(TNF-?) played an important role in weakened bone formation and in increased bone resorption,respectively.Adequate supplementation of vitamin D_3 and reducing serum levels of IL-1?,IL-6 and TNF-? may be very important for prevention and treatment of hepatic bone disease.

Objective Cyclin D1 gene plays a significant role in regulating cell cycle progression.It is reported that over-expression of cyclin D1 gene is intimately associated with origination,development and prognosis of tumor and is associated with tumor cells resistance to chemotherapy drug.Suppression of cyclin D1 protein expression leads to cellular chemosensitization.This study was to determine whether this effect also existed in chronic leukemia cell line K562 by inhibiting the expression of cyclin D1 protein through RNA interference.Methods Plasmid vectors expressing small hairpin RNA (shRNA) targeting at cyclin D1 gene were constructed and transfected into K562 cells by chitosan.Cyclin D1 protein was examined using Western Blotting analysis.The cell cycle and apoptosis were determined by flow cytometry.Cellular chemosensitization was evaluated by MTY assay.Results Expression of cyclin D1 protein was markedly down-regulated after transfection with pshRNA-419 and pshRNA-575 at 48 h.Down-regulation of cyclin D1 protein could affect the redistribution of cell cycle,induce apoptosis of K562 cells,decrease 50％inhibitoryconcentration (IC50) of adriamycin and enhance cellular chemosensitization.But there had no above biological effects observed after transfection with blank vector and control vector of m-pshRNA-790 at 48 h.ConclusionK562 cells could be chemosensitized by the down-regulation of cyclin D1 expression through RNA interference.