BACKGROUND: To avoid acute rejection,it is necessary to use imunosuppressive drug regimen for long term to control immune state.However,imunosuppressive drug regimen of allogenic organ transplantation increases infection incidence of recipients,and induction of allograft immunological tolerance might be an ideal method for solving these problems.The long-term immunologic tolerance has been able to be induced in the experimental rodent models.Among these protocols,donor bone marrow cell (DBMC) infusion exerts an important role in the induction of allograft immunological tolerance.OBJECTIVE: To investigate effects of combination of cyclosporine A (CsA) with DBMC infusion on heterotopic rat cardiac allograft survival time.DESIDN: A randomized controlled animal experiment.SETTING: Renal Disease Center,First Affiliated Hospital of Zhejing University School of Medicine.MATERIALS: This study was performed at the Laboratory Animal Center,Zhejiang University School of Medicine between March 2002 and December 2005.Inbred male Lewis rats (n=40,serving as donors) and male BN rats (n=60,serving as recipients) of SPF grade were used in this study.The protocol was approved by the Hospital's Ethic's Committee.METHODS: Forty rats prepared for heterotopic rat cardiac allograft were randomly divided into 4 groups,with 10 rats in each: control group,in which,rats received no treatment,CsA group,in which,rats received CsA infusion for 7 days successively; CsA +DBMC group,in which,rats received DBMCs during and 6 days after the surgery and additional 7 successive days of CsA infusion,and a DBMC group,in which,rats received DBMCs infusion during and 6 days after the surgery.In addition,BN rats that received beterotopic rat cardiac allograft served BN controls.The survival time of heteroropic rat cardiac allograft was investigated.Serum interleukin-2 level and tumor necrosis factor-α mRNA expression level in the transplanted cardiac allograft were measured. The percentage of antigen presenting cells (APC) from donor,CD3+CD25+ cells,CD4+CD25+ cells,CD86+ cells,and the ratio for CD4+CD45RC+ and CD4+CD45RC- in the recipient peripheral blood karyocytes were measured by flow cytometry 6,12 and 18 days after surgery.MAIN OUTCOME MEASURES: The survival time of beteruropic rat cardiac allograft,serum interleukin-2 (IL-2)level,tumor necrosis factor- α (TNF- α ) rnRNA expression level, rejection grading,the percentage of DBMCs in the recipient peripheral blood karyocytes,CD3+CD25+ cells,and CD4+CD25+ cells,as well as CD86 expression,and the ratio for CD4+CD45RC+ and CD4+CD45RC.RESULTS: Forty Lewis male rats and sixty male BN rats were all included in the final analysis. The heterotopic rat cardiac allograft survival time was longer in the CsA +DBMC group than in the control group and DBMC group (P ＜ 0.05). Serum IL-2 level and TNF- α mRNA expression were respectively lower in the CsA +DBMC group than in the control group and DBMC group ( P ＜ 0.05).The rejection was milder in the CsA +DBMC group than in the remaining 3 transplantation groups.In the CsA +DBMC group,CD 86 expression in the recipient peripheral blood karyocytes was markedly inhibited,and 6 and 12 days after surgery,the ratio for CD4+CD45RC+ and CD4+CD45RC- and the percentage of CD3+CD25+ were respectively lower compared to control group and DBMC group.DBMCs in the recipient peripheral blood karyocytes were more in rats that received DBMC infusion compared to rats that received no BDMC infusion.CONCLUSION: Short-term CsA treatment combined with DBMC infusion can lower acute rejection of heterotopic rat cardiac allograft and prolongssurvival time of cardiac allograft.

Objective:To construct a p53-fused dual luciferase reporter and to test whether this reporter can mimic wild-type p53 activities in a high-throughput screen.Methods:A restriction endonuclease site was added to each terminus and the stop codon of the wild-type full-length p53 open reading frame (ORF) was removed by PCR. A restriction endonuclease site was added to each terminus and the start codon of the ifrelfy luciferase ORF was removed by PCR. The two modified ORFs were inserted upstream of the IRES-induced renilla luciferase ORF in a CMV-derived vector. hTe p53 fusion protein was expressed in cells to test its MDM2-mediated degradation, subcellular localization, and induction of p53-responsive promoter.
Results:hTe p53-fused dual luciferase reporter was successfully constructed. Atfer transfection into the host cells, the reporter expressing the p53 fusion protein that was degraded by oncoprotein MDM2, was mainly located inside the nucleus, and induced the p53-responsive promoter, respectively.
Conclusion:hTe p53-fused dual luciferase reporter (p53FL/IRES/RL) can identify modulators of P53 protein level in a high-throughput screen of genetic or chemical libraries.

Objective To investigate the safety and efficacy of endoscopic tissue adhesive injection combined with sequential endoscopic variceal ligation for gastroesophageal varices of Le,g type. Methods Twenty-three patients with gastroesophageal varices of Le,g type were enrolled to General Hospital of PLA from May 2013 to March 2015, who were treated with endoscopic tissue adhesive injection in the fundic and cardiac site in the first session, followed with endoscopic variceal ligation for esophageal varices. The clinical data, procedure complications and efficacy were retrospectively analysed. Results All procedures were successfully performed with no such evident complications as intraoperative and postoperative bleeding, embolization, mediastinal infection or death with an average hospitalization time of 15. 3±4. 09 days. Mild and moderate thoracalgia occurred in 13 patients(56. 5%), low-grade fever in 2 patients(8. 7%, recovered after symptomatic treatment for 1-2 days) . During the follow-up of 2 weeks, the rate of varices disappearance was 56. 5% (13/23) and no recurrent bleeding was observed. Six months after discharge, 10 patients underwent endoscopy again, varices disappeared in 4 and 6 with remains;the 13 others showed no hemorrhage according to follow-up call. Conclusion The therapy of endoscopic tissue adhesive injection with sequential endoscopic variceal ligation for gastroesophageal varices of Le,g type is safe and efficient.

Objective To evaluate the serum trough concentration and the pharmacokinetics/pharmacodynamics (PK/PD)of vancomycin in patients with severe acute pancreatitis (SAP), and analyze the effect of vancomycin continuous infusion for optimizing the characteristics of its PK/PD.Methods The inhospital patients with SAP received vancomycin treatment and admitted to emergency intensive care unit (EICU) of Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine from January 2011 to December 2016 were enrolled. Steady-state trough concentrations of vancomycin from patients were collected retrospectively. The SAP patients were divided into augmented renal clearance (ARC) and non-ARC groups, as well as systemic inflammatory response syndrome (SIRS) and non-SIRS groups according to the patients with or without symptom above. Adjustments of increased dosage or 24-hour continuous infusion or increase vancomycin dose were made for patients if the steady-state trough concentrations fell below the target level. Steady state trough concentration for vancomycin intermittent infusion or steady state concentration for vancomycin continuous infusion was determined by the fluorescence polarization immunoassay method. PK parameters of vancomycin were calculated using the Bayesian estimator and the area under the serum drug concentration-time curve (AUCc-t), the minimum inhibitory concentration (MIC) and AUCc-t/MIC was recorded and calculated.Results The steady state trough concentration or steady state concentration from 61 patients with SAP were collected with mean steady state trough concentration of vancomycin of (7.7±4.4) mg/L, which was significantly lower than standard concentration (15 mg/L,P < 0.001). Apparent volume of distribution (Vd) and clearance of vancomycin was (1.06±0.26) L/kg and (8.9±2.8) L/h. The serum steady state trough concentration of vancomycin in ARC group (n = 33) was significantly lower than that in non-ARC group (n = 28; mg/L: 6.7±3.5 vs. 8.2±4.1, P < 0.01), clearance was significantly increased (L/h: 9.8±2.9 vs. 7.7±2.2,P < 0.01). Compared with non-SIRS group (n = 31), the serum steady state trough concentration of vancomycin in SIRS group (n= 30) was significantly lowered (mg/L: 6.1±3.2 vs. 13.0±4.2,P < 0.01), and clearance was significantly increased (L/h: 9.4±2.0 vs. 7.1±2.1,P < 0.05). Compared with the only increasing vancomycin dose group (n = 29), vancomycin continuous infusion for 24 hours (n = 21) could significantly reduce daily dosage (mg/kg: 13.6±3.9 vs. 19.1±3.5,P < 0.01), increase the serum trough concentration (mg/L: 18.1±7.0 vs. 12.6±5.3,P < 0.01), and improve the AUCc-t/MIC.Conclusions The serum trough concentration of vancomycin was significantly reduced in SAP patients with ARC. The more serious of the SIRS is, the lower the vancomycin trough concentration is. Vancomycin 24-hour continuous infusion could optimize the PK/PD parameters, decrease the daily dose, increase the clinical effect, and reduce the bacterial resistance.

Objective To explore the clinical characteristics and outcome of group B streptococcus (GBS) induced neonatal meningitis and to provide the guide for early diagnosis and appropriate treatment.Methods A retrospective chart review was performed.A total of 19 cases of neonatal purulent meningitis caused by GBS and 22 cases of neonatal purulent meningitis caused by Escherichia coli were identified in the NICU of Guangzhou Women and Children's Medical Center from Nov 1,2011 to Apr 31,2014.The clinical features,treatments and clinical turnover were analysed.Results GBS meningitis accounted for 24.7％ (19/77) of total bacterial positive cultures of blood or cerebral spinal fluid.The average time of progression to early-onset GBS meningitis of 6 early-onset cases mainly complaining of anhelation and groan,was (11.80 ± 11.34)h,and 83.3％ present within 24 hours;the main initial clinical symptoms of 13 late-onset cases[mean age (17.85 ± 7.77) d] were fever.Peripheral blood C-reactive protein concentration of GBS meningitis was significantly higher than that of Escherichia coli meningitis [(154.43 ± 88.64) mg/L vs.(67.52 ± 64.23) mg/L,P =0.001].Compared with Escherichia coli meningitis,the average length of stay in hospital and the recovery time of abnormal cerebral spinal fluid in neonates with GBS infection were both extended by more than 10 days.Conclusion The clinical manifestations of neonatal purulent meningitis caused by GBS are usually non-specific.It is associated with longer hospitalization and recovery time of abnormal cerebral spinal fluid.Antepartum prophylaxis,early diagnosis and therapy are vital for reducing the incidence of complications and mortality of neonatal GBS purulent meningitis.

Objective To investigate epidemiologically the prevalence of obesity and overweight in Mongolian and Han residents aged over 55 years old in pastoral area of Inner Mongolia,China.Method From 2008 to 2009,with diagnostic criteria of overweight and obesity adopted by Chinese and World Health Organization (WHO),an epidemiological investigation was carried out in 9 146 subjects.Result The incidences of obesity and overweight in Mongolian and Han residents were 32.43％ (32.25％ by WHO criteria),19.09％ (9.91％),and 33.60％ (29.85％),15.19％ (7.66％),respectively.The prevalence of obesity in Mongolian residents was higher than that in Han residents (x2 =16.272,P＜0.01).The status of obesity in Mongolian and Han female residents was more marked than that in male residents (P ＜ 0.05).Overweight between male and female of Han population was different(x2 =5.541,P =0.019).The prevalence of obesity between Mongolian and Han was statistically different (x2 =16.272,P＜0.01).Waist circumference,waist/height ratio,and body mass index between Mongolian and Han were also different (all P ＜ 0.01).Conclusion Differences in prevalence of overweight and obesity were found between Mongolian and Han ethnics among residents aged over 55 in pastoral area of Inner Mongolia,China.

Objective To evaluate the effect of electroacupuncture (EA) pretreatment on NODlike receptor pyrin domain-containing 3 (NLRP3) in neurons during cerebral ischemia-reperfusion (I/R) in rats.Methods Fifty-four adult male Sprage-Dawley rats,aged 7 weeks,weighing 250-280 g,were randomly divided into 3 groups (n =18 each) using a random number table:sham operation group (group S),group I/R,and EA pretreatment group (group E).Cerebral I/R was induced by occlusion of the right middle cerebral artery for 90 min using a nylon thread inserted into the internal carotid artery and advanced intracranially to block the blood flow,followed by reperfusion.In group E,the acupoint Baihui was stimulated with an electric stimulator (sparse-dense wave,frequency 2 Hz/15 Hz,intensity ≤ 1 mA) for 30 min once a day for 5 consecutive days,and the model of cerebral I/R was established at 24 h after the last stimulation.At 72 h of reperfusion,neurological function was assessed and scored.The rats were then sacrificed,and their brains were removed for determination of cerebral infarct volume (using TTC staining),expression of NLRP3,caspase-1 and interleukin-1beta (IL-1β) in brain tissues (by Western blot),and expression of NLRP3 protein in neurons (by immunofluorescence histochemistry).The percentage of cerebral infarct volume was calculated.Results Compared with group S,the percentage of cerebral infarct volume and neurological scores were significantly decreased,and the expression of NLRP3,caspase-1 and IL-1β in brain tissues was significantly up-regulated in group I/R (P＜0.05).Compared with group I/R,the percentage of cerebral infarct volume and neurological scores were significantly increased,and the expression of NLRP3,caspase-1 and IL-1β in brain tissues was significantly down-regulated ingroup E (P＜0.05).Conclusion The mechanism by which EA pretreatment reduces inflammatory responses during cerebral I/R injury may be related to down-regulation of NLRP3 expression in neurons in rats.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Diseases ; diagnosis ; Bile Duct Neoplasms ; diagnosis ; Cholangiopancreatography, Endoscopic Retrograde ; Cytodiagnosis ; Cytological Techniques ; methods ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Diseases ; diagnosis ; Pancreatic Neoplasms ; diagnosis

Choristoma ; pathology ; Fallopian Tube Diseases ; pathology ; Fallopian Tubes ; pathology ; Female ; Humans ; Uterus

Objective To observe the change of the serum trough concentration and its pharmacokinetics of vancomycin in patients with severe acute pancreatitis (SAP), and to analyze the factors influencing vancomycin concentration. Methods A retrospective analysis was conducted. Steady-state trough concentrations of vancomycin from patients (18-80 years old) with SAP concomitantly with G+ infection admitted to Intensive Care Unit (ICU) of Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine from January 2010 to December 2016 were enrolled. According to the usage time of vancomycin, the patients with SAP were divided into early group (onset within 21 days), middle group (onset between 21-28 days) and late group (onset over 28 days). The gender, age, body weight, clinical diagnosis, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ ) score, renal function, and the pharmacokinetic parameters were recorded. Influencing factors of vancomycin was analyzed by multiple linear regression and stepwise regression. Results Fifty-eight patients were enrolled who contained 134 times trough concentrations of vancomycin. There were 41 patients enrolled and 61 times of trough concentrations in the early group, 24 patients enrolled and 33 times of trough concentrations in the middle group, and 28 patients enrolled and 40 times of trough concentrations in the late group. There was no significant difference in gender, age, body weight, serum creatinine, creatinine clearance (CCr), albumin, APACHE Ⅱ score among the three groups. There was significantly difference in the duration from the onset time to vancomycin administration between early, middle groups and late group (days:15.9±3.2, 23.3±2.2 vs. 35.0±6.7, both P < 0.05). The positive liquid balance in early group was lower than that of late group (mL: 1565.2±3132.1 vs. 3675.1±3411.5, P < 0.01), while it was increased in the middle group as compared with that of late group (mL: 5078.7±3892.4 vs. 3675.1±3411.5, P < 0.05). The average daily dose of vancomycin in the early, middle and late groups were (14.7±5.0), (15.0±2.8), (17.0±4.2) mg/kg, respectively, and there was no significant difference (P > 0.05). Compared with the standard concentration (15 mg/L) of vancomycin, the serum trough concentration of vancomycin was significantly reduced in SAP patients [(7.5±4.3) mg/L, P < 0.01]. Apparent volume of distribution (Vd) was (72.4±15.4) L, and clearance rate (CL) was (9.0±2.8) L/h. According to the Bayesian, the serum trough concentration of vancomycin was significantly reduced in early group and middle group compared with late group (mg/L: 5.0±2.1, 7.3±2.5 vs. 11.5±5.1, both P < 0.01), CL was significantly increased (L/h: 10.5±3.0, 8.1±1.9 vs. 7.4±1.9, both P < 0.05), and Vd was significantly increased in early group compared with late group (L: 73.7±15.5 vs. 71.0±12.6, P < 0.05). It was shown by multiple linear regression analysis that there was strong relationship between serum trough concentration and the serum creatinine, CCr, average daily dose and the starting time of vancomycin treatment (r value were 0.449, -0.318, 0.373, 0.763, respectively, all P < 0.05). Conclusions The serum trough concentration of vancomycin was significantly reduced in SAP patients. And the earlier usage of vancomycin, the lower of the trough concentration is. Therefore, higher dosage regimen was needed to ensure the clinical effect, and reduce the bacterial resistance.