Objective To explore the effect of high bilirubin on myocardial injury.Methods The levels of serum total bilirubin (TBIL),creatine kinase (CK),creatine kinase-isoenzyme MB(CK-MB),cardiac troponin Ⅰ (cTnI) were measured and compared in newborns without obvious physiological jaundice (control group) and newborns with hyperbilirubinemia (study group) before and after treatment.Results Compared with those in control group,the levels of serum TBIL,CK,CK-MB and cTnI in study group before treatment were obvious higher[(270.24 ± 36.89) μ mol/L vs.(68.95 ± 10.87) μ mol/L,(414.27 ± 135.45)U/L vs.(178.35 ± 59.62) U/L,(63.31 ± 23.25) U/L vs.(16.78 ± 8.89) U/L,(0.39 ± 0.11) μ g/L vs.(0.18 ± 0.07) μ g/L],and the differences between two groups were statistically significant (P ＜ 0.05).The levels of serum TBIL,CK,CK-MB and cTnI after treatment in study group [(69.13 ±9.98) μmol/L,(183.35 ± 61.01) U/L,(17.45 ± 9.0.8) U/L,(0.19 ± 0.01) μ g/L] were decreased obviously compared with those before treatment,and the differences between two groups were statistically significant (P ＜ 0.05).There was no significant difference between control group and study group after treatment (P ＞ 0.05).Conclusions High bilirubin can lead to a reversible myocardial injury.Cardiac enzymes and troponin testing should be routinely done for neonatal hyperbilirubinemia.It suggests doctors to attend to the treatment of myocardial protection.

Objective To investigate the predictive value of umbilical cord blood bilirubin for pathological jaundice in healthy term newborns. Methods Two ml navel string vein blood of baby were collected after giving birth in the normal newborn, and the hemobilirubin was detected by accidentally oxidation method. After birth, the infant's bilirubin level was tested on the forehead by the transcutaneous bilirubinometer at 8:00 -9:00 every morning until discharging from hospital. The ration of pathological jaundice of newborn and its treatment were analyzed in different levels of cord blood hemobilirubin. Results Fifty-nine cases ( 22.96% ) with pathological jaundice were diagnosed in 257 newboms.The concentration of cord blood hemobilirubin in baby with pathological jaundice [(39.68 ±8.10) μmol/L] was significantly higher than that of the normal newborn [(30.05 ±5.51) μmol/L](P＜0.01). As the concentration of cord blood hemobilirubin was increased, the incidence of pathological jaundice was raised (P＜ 0.01), and the cases that needed to intervention treatment was increased(P＜ 0.01). Conclusion The detection of the level of cord blood hemobilirubin is not only very worthy to estimate the occurrence of pathological jaundice of newborn, but also offer reliable evidence for clinical early diagnosis and treatment.

Objective To investigate the diagnostic value of plasma cardiac troponin Ⅰ (cTnI) and brain natriuretic peptide (BNP) for heart failure complicated with pneumonia in children.Methods 30 children of heart failure complicated with pneumonia (heart failure group),30 children with pneumonia in acute phase (pneumonia group) and 30 healthy children (control group) were selected.The levels of plasma cTnI and BNP in the heart failure group were detected in acute phase(6-24h) and recovery phase(5-7d),and the results were compared with pneumonia group and control group.Results The levels of plasma cTnI and BNP in the heart failure group were significantly higher than those in the pneumonia group and control group(F =5.303,4.632,all P ＜ 0.05).The levels of plasma cTnI and BNP were not significantly different between the pneumonia group and control group(P ＞ 0.05).The levels of plasma cTnI and BNP in recovery phase of the heart failure group were significantly lower than those in acute phase of the heart failure group (t =4.291,5.393,all P ＜ 0.05),but there were no siganificant differences compared with the pneumonia group and control group(P ＞ 0.05).Conclusion The plasma cTnI and BNP maybe a reliable index in the diagnosis of heart failure complicated with pneumonia in children.

BACKGROUND: Recombinant adeno-associated virus 2(rAAV-2) has attracted considerable attention due to its nonpathogenic nature in contrast to other viral vectors such as adenoviral and retroviral vectors in gene therapy attempts.OBJECTIVE: To explore rAAV-2 transduction to bone marrow mesenchymalstem cell(BMSC) in vitro and evaluate the possibility of using rAAV-2 as a vector for gene therapy of acute myelogenous leukemia(AML).DESIGN: An open experiment with cells as the observational subjects.SETTING: Department of Hematology, Nanfang Hospital, Southern Medical University.MATERIALS: The experiment was conducted in the Department of Hematology, Nanfang Hospital, Southern Medical University from February to July 2004. We used passages 3 to 5 BMSCs derived from six de novo AML patients and four healthy volunteers in this study.METHODS: BMSC was isolated from 6 to 10 mL of bone marrow aspirates obtained from the iliac crests of the patients who had been diagnosed as having de novo AML and from those of healthy volunteers. The acquired BMSC was infected by rAAV-2 which contained enhanced green fluorescent protein (rAAV-2-eGFP) at different multiplicity of infection(MOI) (MOI = 1 × 102,1 × 103, 1 × 104, 1 × 105, 1 × 106, 1 × 107) . Then we observed through phase contrast fluorescent microscope and flow cytometer to evaluate green fluorescent protein(GFP) expression 10 to 14 days after transduction. GFP expression was observed as the rAAV-2-eGFP transduced BMSC cultured in vitro. We also observed the in vitro gene expression profile of GFP in rAAV-2-eGFP transduced BMSC which was selected by neomycin ( G418). First, we confirmed GFP expression in BMSC through phase contrast fluorescent microscope, then on flow cytometer to detect the percentage of GFP expression.MAIN OUTCOME MEASURES: The efficiency of rAAV-2-eGFP transduction to BMSC. GFP expression was observed through phase contrast fluorescent microscope and flow cytometer at different time points after transduction.rAAV-2-eGFP to BMSC derived from normal volunteers and AML patients had no significant differences. GFP began to express 10 to 14 days after transduction, and the transduction efficiency ranged from 0. 3% to 1.4%. By changing infection condition, we could not make a higher transduction efficiency( P ＞ 0.05) . One round infection of BMSC by rAAV-2-eGFP at a MOI of 1 × 105 was ( 1. 030 ± 0. 034) %, 3 rounds of infection of BMSC by rAAV-2-eGFP at a MOI of 1 × 105 was (1. 140 ±0. 036)%, and coinfected by LipofectAMINE was (1. 380 ± 0. 054)%. However, 293 cell line which was the package cell of rAAV-2 could be efficiently transduced by AML patients transduced by rAAV-2-eGFP at MOI = 1 × 105: The percentage of GFP expression cell gradually decreased from 1.14% at day 12 after transduction to 0. 6% as cell passaged from 2 to 3, and maintained at a level of 0. 5% to 0. 6% later on till 61 days after transduction. After selected by neomycin(G418) 1 month later, rAAV-2-eGFP transduced BMSCs could maintain a long-term GFP expression at a level of 6.0% in vitro without significant decay within 100 days of observation period after transduction.CONCLUSION: The advantages of rAAV-2 mediated gene transduction lie in safety, no immune response to the host, and long-term expression maintained by the target gene. rAAV-2 and BMSC can be used for in vitro gene therapy, and as a systemic gene delivery system, it might be an alternative for systemic gene therapy in the future.

Objective To investigate the clinical featurcs of hepatic perivascular epithelioid cell carcinoma (PEComa) and the experiments in the surgical treatment of PEComa.Methods The clinical data of 16 patients with hepatic PEComa who received surgical treatment at the First Affiliated Hospital of Nanjing Medical University from January 2008 to January 2012 were retrospectively analyzed.The incidence,clinical manifestations,imagiong characteristics,surgical outcomes and pathological manifestations of this disease were analyzed.B sonography,hepatic function test,tumor markers test and epigastric computed tomography (CT) were applied to detect tumor recurrence and metastasis.The follow-up was ended in December 2012.Result Middle aged and female patients took large part of the patients.Of the 16 patients,8 had no subjective symptoms,and other patients had discomfort in the right upper quadrant,pain or tenderness of the liver.All the tumors were solitary,and most of them located at the right liver (11 tumors were in the right liver,4 in the left liver,1 in the caudate lobe).No specific features were detected by preoperative B sonography,while inhomogeneous low density in the tumor region was detected on by CT.All the patients received partial liver resection without morbidity and morality.The results of immunohistochemistry showed that thc expressions of HMB-45,Melan-A and vascular smooth muscle actin were positive.The mean time of follow-up was 27.9 months (range,9.0-46.0 months),no tumor recurrence and death was observed during the follow-up.Conclusions Middle aged females are susceptive to hepatic PEComa,and patients have no specific clinical presentations.Preoperative CT examination is helpful for differential diagnosis of PEComa,and partical hepatectomy can achieve satisfactory short-term clinical outcomes.

Objective To evaluate early enteral nutrition in patients after liver transplantation. Methods This is a prospective, randomized trial consisting of 63 patients divided into two groups to receive TPN and early enteral nutrition respectively for a week. Patients' nutritional status, liver function, nutritional expense and the incidence of postoperative infections and other complications were compared between the two groups. Result Early enteral nutrition after liver transplantation can improve the nutritional condition and it is helpful for the recovery of the liver function. It decreases the rate of postoperative infections and other complications. Conclusions Early enteral nutritional support is feasible alternative to TPN in patients of liver transplantation.

Objective To investigate the clinical significance of detecting cardiac troponin Ⅰ (cTnⅠ) and CK -MB in children with hand-foot-and-mouth disease and myocardial injury.Methods 90 children with hand-foot-and-mouth disease (observation group) were detected the level of serum cTnⅠ and CK-MB.At the same time,40 healthy children were chosen as control group.Results Compared with the control group,the levels of CK-MB and cTnl in observation group were significantly higher than those of the control group (t =8.92,5.46,all P ＜ 0.01),which indicated that children with hand,foot and mouth disease was easier to merge myocardial injury.CK-MB and cTnl levels in high-risk group were significantly higher than those in normal children group (t =9.17,6.13,all P ＜ 0.01),and the levels of CK-MB and cTnl were positively correlated with severe degree (r =0.767,0.683,all P ＜ 0.01).For children with hand,foot and mouth disease merged myocarditis,cTnl diagnostic sensitivity (53.8％) was lower than that of CK-MB diagnostic sensitivity (71.8％),but the specificity was better than that of CK-MB,and the diagnosis of cTnl detection window 2 weeks longer than CK-MB,but after 2 weeks both lose their clinical diagnostic significance.Conclusion Children with hand,foot and mouth disease easily merge myocardial damage,dynamic measuring CK-MB and cTnl levels could help early diagnosis of children with hand,foot and mouth disease whether merged myocardial damage,both applications can also complement each other,more timely and accurate reflection of disease progression and recovery,it is worth promoting.

Objective To investigate the clinical efficacy and prognostic factors of radical hepatectomy of hepatocellular carcinoma (HCC).Methods The retrospective case-control study was conducted.The clinicopathological data of 760 HCC patients who were admitted to the First Affiliated Hospital of Nanjing Medical University from August 2003 to June 2015 were collected.Surgical procedures were determined according to the location,number and size of tumors and anatomical relations among vessels.Observation indicators included:(1)intra-and post-operative situations:surgical procedures,operation time,volume of intraoperative blood loss,cases of intraoperative blood transfusion,postoperative complications,duration of postoperative hospital stay and pathological examination;(2) follow-up:1-,3-,5-year overall and tumor-free survival situations;(3) prognostic factors analysis of HCC patients.Follow-up using outpatient examination and telephone interview was performed to detect patients' survival up to January 2016.Measurement data with normal distribution were represented as-x±s.The survival curve and survival rate were respectively drawn and calculated by the Kaplan-Meier method.The univariate analysis and multivariate analysis were done using the COX regression model.Results (1) Intra-and post-operative situations:all the 760 patients underwent successful operations,including 419 undergoing anatomical hepatectomy and 341 undergoing non-anatomical hepatectomy.R0 and R1 resections were respectively applied to 742 and 18 patients.Two patients were combined with portal vein resection and reconstruction and 1 was combined with resection and reconstruction of inferior vena cava.Operation time,volume of intraoperative blood loss and cases of intraoperative blood transfusion were (226± 115) minutes,(714±706) mL and 88,respectively.Fifty-five patients had postoperative complications,including 20 with abdominal effusion or abscess,16 with pleural effusion,9 with recurrent fever,8 with incisional infection,7 with intra-abdominal hemorrhage,6 with liver failure,3 with pyloric or intestinal obstruction and 2 with renal failure (some patients with multiple complications).Of the 55 patients with postoperative complications,7 with hemorrhage underwent reoperation or interventional therapy and other patients underwent conventional symptomatic treatment.Of 55 patients,5 patients died and other 50 patients were improved.Duration of postoperative hospital stay was (14±6) days.There were 457 patients with minimum margin of tumors ≤ 1.0 cm and 303 with minimum margin of tumors ＞ 1.0 cm.(2) Followup:all the 760 patients were followed up for 1-139 months,with a median time of 25 months.The overall and tumor-free median survival times were 59 months and 31 months,respectively.The 1-,3-,5-year overall and tumor-free survival rates were 81.7％,63.4％,47.9％ and 68.7％,44.9％,29.6％,respectively.(3) Prognostic factors analysis of HCC patients:results of univariate analysis showed that clinical symptoms,alpha-fetoprotein (AFP),Barcelona clinic liver cancer staging,surgical procedures,intraoperative blood transfusion,minimum margin of tumors,number and diameter of tumors,tumor capsule,tumor differentiation,vascular cancer embolus,macrovascular invasion and tumor staging of American Joint Committee on Cancer (AJCC) were related factors affecting prognosis of HCC patients after radical hepatectomy [HR =1.39,1.50,1.92,0.65,1.45,1.68,1.96,1.66,2.26,1.50,2.68,3.37,2.00,95％ confidence interval (CI):1.08-1.79,1.16-1.94,1.68-2.20,0.50-0.84,1.04-2.02,1.28-2.20,1.54-2.49,1.42-1.94,1.69-3.02,1.22-1.85,1.99-3.60,2.61-4.36,1.77-2.27,P＜0.05].Results of multivariate analysis showed that AFP,number and diameter of tumors,tumor differentiation and tumor staging of AJCC were independent factors affecting prognosis of HCC patients after radical hepatectomy (HR=1.61,1.62,1.31,1.40,1.78,95％CI:1.14-2.26,1.22-2.14,1.06-1.63,1.10-1.79,1.27-2.51,P ＜ 0.05).Conclusions The anatomical and non-anatomical hepatectomies are safe and feasible for optional HCC patients,with a good long-term outcome.AFP,number and diameter of tumors,tumor differentiation and tumor staging of AJCC are independent factors affecting prognosis of HCC patients after radical hepatectomy.

ObjectiveTo investigate the therapeutic effects of the conditioning regimen with or without total body irradiation on allogeneic hematopoietic stem cell transplantation in acute leukemia.Methods We retrospectively evaluated clinical outcomes in 287 allo-HSCT recipients with acute leukemia (ALL- 105,AML-129,and AUL-53) who received myeloablative conditioning regimen with or without total body irradiation from January 2002 to August 2011.Two hundred and six patients obtained complete remission (CR) and 81 non-remission (NR) before transplantation.One hundred and ninety-nine patients received conditioning with total body irradiation (TBI+ Cy group,9 Gy given over 2 days),and 88 patients received busulfan (BuCy group,3.2 mg·kg-1 ·d-1 ),both followed by cyclophosphamide.ResultsThere were no statistically significant differences in hematopoietic reconstitution,regimen-related toxicity (RRT),graft-versus-host disease (GVHD) and relapse between two groups.For patients with AML and AUL,there was no significant difference in the 5-year survival between the two regimens (P＞ 0.05),while for ALL-CR patients,the TBI + Cy regimen had a higher over survival rate (52.0％ vs.31.3％,LogRank=4.249,P＜0.05) and DFS (50.4％ vs.27.8％,LogRank =4.445,P＜0.05) than BuCy.In TBI + Cy group and BuCy group,the proportion of CD19+ B cells at the first month after HSCT was (4.04 ± 1.86)％ and (1.47 ±0.99) ％ (P＜0.05),that of NK cells at 12th month after HSCT was (23.38 ± 12.19) ％ and (13.11± 7.99) ％ (P＜0.05),and that of CD4+ CD45RO+ cells at 9th month after HSCT was (14.63 ±6.17)％ and (9.07 ± 3.12)％ (P＜0.01),respectively.ConclusionUsing TBI-containing regimen was more effective for treating ALL-CR patients than busulphan-containing regimen,but no difference was found for long-term outcomes in patients with AML and AUL between the two regimens.The 9 Gy TBI-based regimens may not affect recipients' thymic function,T-cells reconstitution and immune tolerance,coming out a non-increase of GVHD.

Background::Heterotaxy (HTX) is a thoracoabdominal organ anomaly syndrome and commonly accompanied by congenital heart disease (CHD). The aim of this study was to analyze rare copy number variations (CNVs) in a HTX/CHD cohort and to examine the potential mechanisms contributing to HTX/CHD.Methods::Chromosome microarray analysis was used to identify rare CNVs in a cohort of 120 unrelated HTX/CHD patients, and available samples from parents were used to confirm the inheritance pattern. Potential candidate genes in CNVs region were prioritized via the DECIPHER database, and PNPLA4 was identified as the leading candidate gene. To validate, we generated PNPLA4-overexpressing human induced pluripotent stem cell lines as well as pnpla4-overexpressing zebrafish model, followed by a series of transcriptomic, biochemical and cellular analyses. Results::Seventeen rare CNVs were identified in 15 of the 120 HTX/CHD patients (12.5%). Xp22.31 duplication was one of the inherited CNVs identified in this HTX/CHD cohort, and PNPLA4 in the Xp22.31 was a candidate gene associated with HTX/CHD. PNPLA4 is expressed in the lateral plate mesoderm, which is known to be critical for left/right embryonic patterning as well as cardiomyocyte differentiation, and in the neural crest cell lineage. Through a series of in vivo and in vitro analyses at the molecular and cellular levels, we revealed that the biological function of PNPLA4 is importantly involved in the primary cilia formation and function via its regulation of energy metabolism and mitochondria-mediated ATP production. Conclusions::Our findings demonstrated a significant association between CNVs and HTX/CHD. Our data strongly suggested that an increased genetic dose of PNPLA4 due to Xp22.31 duplication is a disease-causing risk factor for HTX/CHD.