Obgective To analyze the demographic data,non-specific items,pathogens and antibiotic sensitivity between the children with early-onset and late-onset sepsis,in order to guide the diagnosis and treatment of neonatal sepsis.Methods Three hundred and fifty-two cases with positive blood culture were retrospectively recruited and divided into an early-onset group and a late-onset sepsis group according to the onset of sepsis.Results Of 352 cases,144 cases (40.91％) were the early-onset children while 208 cases (59.09％) were the late-onset children,and in the late-onset group,108 cases occurred due to nosocomial infection.Most neonates of the early-onset term were term infants [107/144 cases (74.31％)],while the preterm infants [77/208 cases (37.02％)] and low birth weight infants[70/208 cases(33.65％)] accounted for the majority of the late-onset group.The asphyxia,perinatal intrauterine distress,meconium-staining amniotic fluid and premature rupture of fetal membranes ≥ 18 h occurred more frequently in the early-onset group [21/144 cases (14.58％),14/144 cases (9.72％),26/144 cases (18.06％),31/144 cases (21.53％)],respectively,while those in the late-onset group were [17/208 cases (8.17％),9/208 cases(4.33％),13/208 cases(6.25％),17/208 cases(8.17％)],respectively,there were significant differences (x2 =4.622,3.886,5.950,13.345,all P ＜ 0.05) between 2 groups.In the early-onset group abnormal temperature[72/208 cases(34.62％)vs 30/144 cases(20.83％)],vomiting or abdominal distention[109/208 cases (52.40％) vs 35/144 cases (24.31％)],lethargy [79/208 cases (37.98％) vs 38/144 cases (26.39 ％)] and umbilicalitis or skin pustule [33/208 cases (15.87 ％) vs 11 / 1 44 cases (7.64 ％)] occurred more frequently in late-onset group,and there were significant differences (x2 =7.853,8.763,5.153,5.265,all P ＜ 0.05).Besides,more cases in the late-onset group had elevated immature neutrophil vs total neutrophil count ratio [27/184 cases (14.67％)] and C-reactive protein value [76/206 cases (36.89％)],compared with those in early-onset group [9/133 cases (6.77％),38/143 cases(26.57％)],and there were significant differences (x2 =4.794,4.087,allP ＜ 0.05).Compared with early-onset group,patients in the late-onset group were more likely to suffer from suppurative meningitis [17.79％ (37/208 cases) vs 8.33％ (12/144 cases);x2 =6.348,P ＜ 0.05].In terms of pathogens,the main pathogens in the early-onset group were gram negative bacteria[39.58％ (57/144 cases),including detection of Klebisella pneumoniae in 21 cases and E.coli in 20 cases] and coagulase negative staphylococcus[32.64％ (47/144 cases)].In late-onset group,the main pathogens were gram positive bacteria [58.65％ (122/208 cases)],including detection of coagulase negative staphylococcus in 90 cases(43.27％) and E.coli [17.79％ (37/208 cases)].There was no significant difference in prognosis between 2 groups(x2 =1.187,P =0.552).Conclusions Early-onset sepsis and late onset sepsis differ in the clinical manifestation and laboratory findings.Distinguishing neonatal early-onset and late onset septicemia is of clinical significance in choosing appropriate antibiotics.

Objective To investigate the efficacy of ventilation with selective lobar collapse for thoracic surgery in elderly patients with chronic obstructive pulmonary disease (COPD).Methods Thirty ASA Ⅱ or Ⅲ patients with COPD,aged 65-80 yr,with a body mass index of 16-28 kg/m2,undergoing radical resection of esophagus cancer,were randomly divided into 2 groups (n=15 each):one-lung ventilation (OLV) group and ventilation with selective lobar collapse group (group SLC).In group OLV,OLV was performed,while in group SLC,the balloon of the blocker was placed at 0.5 cm below the opening of the upper lobe bronchus and the lower lobe was collapsed when chest was open.The patients were mechanically ventilated (VT =7-8 ml/kg,RR =14-16 bpm,I∶E=1∶1.5-2.0,FiO2 =100％).Peak pressure (Ppeak),plateau pressure (Pplat),airway resistance (Raw),and dynamic lung compliance (Cd) were measured at 10 min of two-lung ventilation in supine position (T0),at 5,45 and 90 min of OLV or selective lobar collapse (T1-3) and at 10 min of two-lung ventilation in lateral position after the end of operation (T4).Arterial blood samples were obtained at To,T3 and T4 for blood gas analysis.Oxygenation index (OI),alveolar-arterial oxygen gradiant (P(A-a)O2),and respiratory index (RI) were calculated.Results Ppeak,Pplat and Raw were significantly lower at T2-4,while Cd was significantly greater at T1-4,OI was significantly higher at T3,4,and P(A-a)O2 and RI were significantly lower at T3,4 in group SLC than in group OLV.Conclusion The thoracic operation can be completed safely using ventilation with selective lobar collapse and OLV,however,ventilation with selective lobar collapse can improve oxygenation and provides better ventilatory efficacy than OLV in elderly patients with COPD.

Objective To explore the change of copeptin in plasma and its significance in patients with intracerebral hemorrhage combined with stress ulcer.Methods Eighty patients with intracerebral hemorrhage were collected.Forty-nine patients of pure intracerebral hemorrhage and 31 patients of intracerebral hemorrhage combined with stress ulcer were included.Thirty healthy people were taken as controls.The level of copeptin in plasma was measured and compared in all subjects.Results The level of copeptin in plasma in patients with pure intracerebral hemorrhage and intracerebral hemorrhage combined with stress ulcer was significantly higher than that in controls:(303.684 ± 68.691),(527.034 ± 74.111) ng/L vs.(121.460 ± 53.364) ng/L,and the level of copeptin in plasma in patients with intracerebral hemorrhage combined with stress ulcer was significantly higher than that in patients with pure intracerebral hemorrhage.The differences were statistically significant (P ＜ 0.05).Conclusion The level of copeptin in plasma in patients with pure intracerebral hemorrhage increases significantly,and it is much higher in patients with intracerebral hemorrhage combined with stress ulcer.

Objective To investigate the effects of microRNA-196a(miR-196a) inhibitory sequences transfection on HOXB8 expression in PANC1 cells.Methods PANC1 cells were divided into control group,miR-196a inhibitory sequences group and siRNA control group.Liposomal transfection method was applied to transfect miR 196a inhibitory sequences and siRNA control into PANC1 cells.RT-PCR and Western blot were used to detect the expressions of miR-196a and HOXB8 mRNA and protein.Results After miR-196a inhibitory sequences transfection,when compared with that of siRNA control group,the expression of miR-196a was significantly decreased (0.05 ± 0.054 vs.0.839 ± 0.025,t =3.12,P ＜0.05) ; and the expression of HOXB8 mRNA was significantly increased by 1.57 folds (2.20 ± 0.07 vs.1.29 ± 0.10,t =3.86,P ＜ 0.05),the expression of HOXB8 protein was also obviously increased (0.90 ± 0.03 vs.0.40 ± 0.10,t =3.11,P ＜ 0.05).Conclusions MicroRNA-196a down-regulates the expression of HOXB8.

Objective To explore the effect of TNF-α in intracellular Ca~(2+)-overload associated myocardial injury. Methods Thirty Wistar rats were randomly divided into three groups:control group,Ca~(2+)+-paradox group,and pentoxifylline treatment group. The intracellular Ca~(2+)-overload rat model was established by pre-filling the rats with Langendorff for 20 minutes,isolated rat hearts subjected to Ca~(2+)-depletion for 5 minutes and Ca~(2+)-repletion for 30 minites(Ca~(2+)-paradox). Changes in hemodynamics indexes were monitored continuously. TNF-α in cardiac tissues was tested by ELISA method,and nuclear factor-κB(NF-κB)in cardiac tissues was detected with Western blot. Results Dramatic depression in left ventricle contraction function was found in the Ca~(2+)-paradox hearts:significant decrease in left ventricular diastolic pressure(LVDP),markedly elevated left ventricular end diastolic pressure(LVEDP),decreased dP/dt ratio,increased TNF-α content,decreased cytosolic/homogenate NF-κB ratio. All these changes in Ca~(2+)-paradox group were significantly attenuated upon the treatment with 100 μmol/L pentoxifylline. Conclusions Activation of NF-κB and increased production of TNF-α may play important roles in cardiac injury associated with intracellular Ca~(2+)-overload.

Objective To compare the efficacy and safety of E1B gene-deleted adenovirus (H101)and ethanol in treating advanced pancreatic carcinomas by intratumoral injection combined with intravenous gemcitabine.Methods We constructed an orthotopic nude mouse model of pancreatic carcinoma through cancer cell injection into pancreas.A total of 54 nude mice were randomly allocated to 6 groups to accept H101,ethanol or saline (control) intratumoral injection,combined with or without intravenous gemcitabiein.The animals were sacrificed 4 weeks after the treatment and the pancreatic tumors were collected to determine the size,existence of metastasis,distribution of virus by indirect immunofluorescence and apoptosis in tumor by TUNEL and electron microscope.Results All mice completed the scheduled treatment,while 3 died in 48 hours after ethanol injection resulting in a mortality of 16.7％ (3/18).On the contrary,no mice died in the adenovirus injcction group.The average tumor size in group of H101 intratumoral injection combined with intravenous gemcitabie was significant smaller than that in group of saline injection with or without systemic gemcitabie (P =0.008,0.040,respectively).Similar differences were observed between ethanol intratumoral injection and control groups (P =0.012,0.041).Meanwhile,the H101 was absent in all the other organs except the pancreas,which meant that the selectivity of the H101 was tremcndous.The virus combine gemcitabie group had higher apoptosis rate in tumor (83.2 ± 35.7) ％,determined by TUNEL.Conclusion E1B gene-deleted adenovirus intratumral injection in combination with intravenous gemcitabine treating pancreatic carcinomas is efficient and safe,in spite of its lower effectiveness than ethanol.

Objective To investigate the expression of DNA-methyltransferases 3B(DNMT3B)gene in human pancreatic carcinoma and to evaluate its relationship with elinicopathologic parameters.Methods 42 samples of pancreatic carcinoma tissues and 42 para-carcinoma tissues and 10 normal pancreatic tissues were collected and the expression of DNMT3B mRNA and protein Was detected by real.time PCR and immunohistochemistry techniques.Results The expression of DNMT3B mRNA(RQ level)in human pancreatic carcinoma tissues and para-carcinoma tissues,normal pancreatic tissues was 9.4±5.9,1.02±0.71 and 0,respectively,and the difference was statistically significant(P<0.05).The rate of expression of DNMT3B protein in human pancreatic carcinoma tissues,para-carcinoma tissues and normal pancreatic tissues were 83.3%,14.3%and 10%,respectively,and the difference wag also statistically significant(P<0.01).The expression of DNMT3B mRNA correlated significantly with clinical staging,differentiation degree of the tumor and lymph node metastasis(P<0.01 or P<0.05).The expression of DNMT3B protein correlated significantly with the location ofthe tumor and lymph node metastasis(P<0.01 or P<0.05).The expression of DNMT3B mRNA and protein Was not assecimed with age,sex,neural invasion,tumor size,sernm CEA and CA19-9.Conclusions Highly expressed DNMT3B mRNA and protein may indicate the lymph node metastasis and poor prognosis in human pancreatic carcinoma.

Objective To investigate the feasibility of induction of experimental chronic pancreatitis rat model with L-arginine.Methods Animals were randomly divided into control group,arginine 12 h group,arginine 24 h group and arginine 7 d group with 10 rats in each group.L-arginine solution was intraperitoneally injected twice with an interval of 1 h.Serum amylase and glucose levels at corresponding time points were detected and histopathological scores of pancreas were evaluated.Collagen in pancreas was stained with Van Gieson method.Results Serum amylase levels were (1 634±890 ) U/L,( 3 872±2 676 ) U/L,( 3 307±2 197)U/L and (1 561±304) U/L in control group,arginine 12 h group,arglnine 24 h group and arginine 7 d group,respectively.The serum amylase level in arginine 7 d group was significantly lower than those in arginine 12 h group and arginine 24 h group (P ＜ 0.05 ).There was no significant difference in serum glucose level among all the groups.Histopathological scores were 0.8±0.4,5.1±2.6,6.5±2.2 and 4.5±1.6,respectively.The histopathological score of arginine 7 d group was significantly lower than those in arginine 24 h group (P ＜ 0.05 ).Obvious collagen could be found in pancreatic parenchyma in arginine 7 d group,while little collagen was found in pancreatic tissue in control,arginine 12 h and arginine 24 h groups.Conclusions Injection of L-arglnine induced fibrosis in pancreatic parenchyma and proliferation of tubular complex 7 days later,and it could be used for chronic pancreatitis model induction.

Objective To investigate the expression of SUFU protein in human pancreatic carcinoma tissues and to explore the relationship between the expression of SUFU protein and the clinicopathologic parameters.Methods The expression of SUFU protein in 28 samples of pancreatic cancer tissues and 20 adjacent normal pancreatic tissues and 4 normal pancreatic tissues was detected by immunohistochemical method.And the relationship between the expression of SUFU protein and the clinicopathologic parameters were determined.Results SUFU protein was positively expressed in cytoplasm and nucleus of pancreatic carcinoma cells, while it was not expressed in the duct, acinar and islet of tumor-adjacent tissues and normal pancreatic tissues, and difference was statistically significant (P ＜ 0.05).The high SUFU protein expression was related to the clinical stage ( P ＜ 0.05 ), but not the age, gender, tumor location , tumor size, lymph node metastasis and liver metastasis.Conclusions SUFU protein was highly expressed in pancreatic cancer, and SUFU may play certain role in the pathogenesis of pancreatic cancer.

Objective To construct a nucleic acid vaccine expressing H.pylori HpaA and inter- leukin-2 gene and to identify the immunogenicity of the vaccine proteins in vitro and protection in vivo. Methods The H paA gene fragment was amplified by polymerase chain reaction(PCR) from the genomic DNA of the standard H.pylori strain 17874.Mouse interlukin(IL)-2 gene was amplified from pClneo- IL-2.The HpaA and IL-2 were cloned into pUCmT vector.After DNA sequences of the amplified HpaA gene and IL-2 were confirmed,both were cloned into the eukaryotic expression vector pIRES through a serial of enzyme digestion and ligation reactions.The recombinant plasmids were screened by PCR and restriction enzyme digestion.Then,recombinant pIRES-HpsA-IL-2 was transfected to COS-7 cells using Lipofectamine~(TM)2000.The immunogenicity of HpaA and IL-2 protein was detected by SDS- PAGE and Western blot.The recombinant plasmids were transformed to LB5000 and then to final host SL7207.The recombinant strains were passaged repeatedly.The mice were challenged with H.pylori after 4 weeks of inoculation of nucleic acid vaccine.H.pylori infection was detected by rapid urease test.Results The amplified HpaA gene fragment and IL-2 were confirmed by sequence analysis.The eukaryotic expression vector plRES and the pIRES-HpaA-IL-2 construction were confirmed by PCR and restriction digestion.The expressions of HpaA(30 000) and IL-2(14 000)protein by pIRES-HpaA-IL- 2 were detected by Western blot.The in vivo study showed that 75.0% and 58.4% of mice vaccinated by HpaA-IL-2 and HpaA,respectively,were protected anaigst H.pylory infection,which was signifi- cant different in comparison with PBS control (P