Objective To determine the pathogen profile and antibiotic resistance in aerobic isolates from blood cultures of neonates. Methods All blood culture reports (n=28120) from newborns admitted to the Department of Neonatology during 2002-2012 were analyzed, and the sensitivity patterns were recorded. Results A total of 1665 bacteria were isolated from 1606 blood culture-positive samples and the positive rate of blood cultures was 5.7%(1606/28120). Gram-positive bacteria were iso-lated in 1336 cases, with Staphylococcus epidermidis (902 cases) and Staphylococcus haemolyticus (206 cases) being the com-mon bacteria. Klebsiella pneumoniae (108 cases), followed by Escherichia coli (73 cases), were the major Gram-negative bacte-ria (235 cases). The determination of the antibiotic resistance of aerobic isolates was performed in 2012. Most Gram-positive iso-lates were sensitive to vancomycin and moxifloxacin, and more than 90%were resistant to penicillin while most of Gram-nega-tive isolates were sensitive to amikacin and imipenem. Conclusions Staphylococcus epidermidis, Staphylococcus haemolyticus, Klebsiella pneumoniae and Escherichia coli remain to be the principal organisms responsible for neonatal sepsis.

OBJECTIVE To observe the disinfection efficacy of nano-light catalytic air disinfector(Jian Bai Le disinfector) in ophthalmic outpatients operating room.METHODS The efficacy of disinfection of the disinfector in the operating room without and with staff was detected respectively.This efficacy of disinfection was compared with that of ultraviolet ray.RESULTS The average eliminated rate of natural bacteria in the air was 88.9% vs 89.7%,and the average bacterial colony number was 63.9 vs 75.0 CFU/m3,respectively,after disinfection by the disinfector or by ultraviolet ray without staff.When the device worked continuously with staff,Their average bacterial colony number was 26.6,92.2,150.0 and 155.3 CFU/m3,vs 150.2,166.7,355.1 and 738.4CFU/m3 in the period of 30,60,120 and 180 minutes after operation,respectively.CONCLUSIONS The bactericidal efficacy of disinfector is distinctly better than that of ultraviolet ray lamp.

The effects of targeted silencing of heparanase gene by small interfering RNA (siRNA) on invasiveness and metastasis of osteosarcoma cells (MG63 cells) were investigated in the present study. Two complementary oligonucleotide strands were synthesized and inserted into pGenesil-1 vector based on the mRNA sequence of heparanase gene. The expression vector containing short hairpin RNA (pGenesil-shRNA) was constructed successfully. MG63 cells were randomly allocated into 3 groups: blank group, empty vector (pGenesil) transfected group and expression vector (pGenesil-shRNA) transfected group. Under the induction of Lipofectamine 2000, the recombinants were transfected into MG63 cells. Heparanase gene expression level was detected by RT-PCR and Western blotting. Cell proliferation was measured by MTT assay. Cell invasiveness and metastasis were examined by cell adhesion and Transwell-ECM assays. HUVECs migration assay was applied for the detection of angiogenesis. As compared with negative controls, the mRNA and protein expression levels of heparanase were down-regulated by 76.1% (P<0.01) and 75.3% (P<0.01) respectively in the pGenesil-shRNA transfected group. Meanwhile, the proliferation, adhesiveness, invasiveness and angiogenesis properties of MG63 cells were all significantly inhibited. It was suggested that targeted silencing of heparanase gene by siRNA could dramatically inhibit the invasiveness and metastasis of osteosarcoma cells.

Purpose To investigate the role of S3I-201 on tubular interstitial lesion in lupus nephritis.Methods MRt/MpJ mice were designated as the control group.MRL/lpr nice were randomly divided into LN group,S3I-201 group and DMSO group.The serum and 24 h-urine were collected to detect the serum creatinine,blood urea nitrogen and urine protein.Immunohistochemistry was used to detect the expression of FN.Western blotting analysis was used to determine the expression of E-cadherin,α-SMA,MCP-1,ICAM1,STAT3 and p-STAT3.Results Compared with the expression level in control group,the protein level of α-SMA,MCP-1,ICAM1 and FN were increased in renal tissue of MRL/lpr mice,while the expression of E-cadherin was markedly decreased.And the STAT3 was activated in renal tissue of MRL/lpr mice.The administration of S3I-201 could inhibite the activation of STAT3 and ameliorate the expression of E-cadherin,α-SMA,MCP-1,ICAM-1 and FN.Conclusion S3I-201 can relieve the tubular interstitial leison,which maybe concerned with the phosphorylation of STAT3.

To express recombinant carboxypeptidase from Thermus aquaticus (Cpase Taq) in Pichia pastosis, the open reading frame coding thermostable Cpase Taq was optimized based on the preference of P. pastoris codon usage and synthesized in vitro. The novel gene was cloned into P. pastoris expression vector pHBM905A and the sequence coding 6xHis tag was fused with the ORF of Cpase Taq gene. The recombinant plasmid was named pHBM905A-Cpase Taq and transformed into P. pastoris GS 115. Transformants were induced with 1% methanol for 72 h until the enzyme yield reached 0.1 mg/ml. The enzyme was purified and its enzymatic properties were analyzed. The results showed that the specific enzyme activity reached maximum at 75 °C and pH 7.5, which was about 80 U/mg. It was the first report about the secretory expression of Cpase Taq in P. pastoris GS115. Because of its large-scale preparation, this enzyme may be applied in industrial hydrolysis of peptides into amino acids in the future.
Bacterial Proteins ; biosynthesis ; genetics ; Carboxypeptidases ; biosynthesis ; genetics ; Cloning, Molecular ; Codon ; Hydrolysis ; Open Reading Frames ; Pichia ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Thermus ; enzymology

Objective To observe the activation of anti-viral innate immune response of type Ⅰinterferon and inhibition of hepatitis B virus (HBV)genome replication in mice by HBV-3p-siRNA. Methods HBV-3p-siRNA was designed by targeting specific sequence of HBV S/P mRNA and was generated by in vitro transcription.Negative control siRNA (NC-siRNA)and non-modified HBV-siRNA were used as control groups.Blood samples were collected from tail vein of mice and the model of HBV-infected mice were established by hydrodynamic injection.Forty mice were divided into 4 groups with 10 in each group.The model group was only injected with pGL3.0-HBV1 .2 copy plasmid.The negative control group received peritoneal injection of NC-siRNA.HBV-siRNA group received peritoneal injection of HBV-siRNA and HBV-3p-siRNA group received peritoneal injection of HBV-3p-siRNA.The interferon-β(IFN-β)and hepatitis B surface antigen (HBsAg)in serum were detected by enzyme linked immunosorbent assay (ELISA).The copies of HBV DNA were assessed by fluore scence quantitative polymerase chain reaction (PCR ).The statistical difference between groups was determined using One way-ANOVA analysis by LSD or Dunnett T3.Results Serum level of IFN-β was (12.37±5 .32)pg/mL in model group,(22.61 ±6.29 )pg/mL in negative control group,(26.40±5 .39)pg/mL in HBV-siRNA group and (68.37± 21 .00 ) pg/mL in HBV-3p-siRNA group.The secretions of IFN-β into serum were significantly enhanced by HBV-siRNA and HBV-3p-siRNA compared with model group (F =23.988 and 46.523,respectively,both P <0.01).Serum level of HBsAg was (2 864.86±907.11 )ng/mL in model group,(2 198.86±456.89 )ng/mL in negative control group,(1 049.71 ± 396.28 )ng/mL in HBV-siRNA group and (640.86±383.08)ng/mL in HBV-3p-siRNA group.The expressions of HBsAg were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F = 23.537 and 39.144, respectively;P =0.025 and 0.010,respectively).Serum level of HBV DNA was (2.54 ×104 ±1 .46 × 104 )copy/mL in model group,(2.22×104 ±2.62×103 )copy/mL in negative control group,(3.59×103 ±2.88×103 )copy/mL in HBV-siRNA group and (2.65 ×103 ±1 .46×103 )copy/mL in HBV-3p-siRNA group.Serum level of HBV DNA were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F =15 .013 and 16.741 ,respectively,both P <0.05 ).All of the indicated siRNA used in the experiments showed no apparent effects on the body mass index of the mice models.Conclusion HBV-3p-siRNA,which induces the production of IFN-β and inhibits HBV replication through gene silencing in vivo ,may be a powerful bifunctional antiviral molecule.

10.3969/j.issn.1008-9691.2013.03.009

Objective To study the expression of Ca-A/K channel-related molecules glutamate receptor 2 and 1(GluR2/1) in hippocampus tissues of neonatal rats with hypoxic-ischemic brain damage (HIBD). Methods A total of 60 7-day-old Sprague-Dawley rats were randomly divided into sham operation group and HIBD group. Hippocampal tissues were obtained at 0 h, 1 h, 6 h, 24 h, 48 h and 72 h after HIBD. The expression of GluR2, GluR1 and autophagy marker protein Beclin-1, LC3 were detected by Western blot assay. Results Edema and focal softening and necrosis were observed 6 h after HIBD in the brains of neonatal rats. Compared with Con group, at each time point, the expression levels of GluR2 were lower while the levels of GluR1, Beclin-1 and LC3 were higher significantly in HIBD group (P<0.05). The protein levels of LC3, Beclin-1, GluR1 and GluR2 in hippocampus tissues of HIBD group were significantly different among different time points after the estab-lishment of HIBD model (F=10.65~701.14, P<0.01). The protein level of GluR2 was decreased from 1 h to 24 h after HIBD and reached the lowest level at 24 h. The levels of GluR1, Beclin-1 and LC3 were increased at 6 h, plateaued at 24 h and remained there until 48 h. The levels of these proteins returned back to the initial level at 72 h. Conclusions Ca-A/K channel-related mol-ecules GluR2 and GluR1 play important roles in the autophagic cell death of hippocampus tissues in neonatal rats with hypoxic-ischemic brain damage.

OBJECTIVE To investigate the countermeasures of preventing endophthalmitis in the process of large number of successive cataract phacoemulsification surgeries at the Lifeline-Express train.METHODS All measures of preventing postoperative endophthalmitis during and after cataract surgery procedures from Mar 2007 to Aug 2007 at the Second Lifeline-Express train were retrospectively analyzed and summarized.RESULTS There was not any postoperative endophthalmitis occurred from 2539 procedures of cataract phacoemulsification surgeries.CONCLUSIONS A series of the effective approaches as muticulous as possible are keys to prevent the postoperative endophthalmitis in the process of large number of successive cataract phacoemulsification surgeries.

This study was aimed to investigate the efficacy ofTong-Bu San-Sheng(TBSS) Decoction to reduce the toxicity and side effects of chemotherapy, as well as prolong progression-free survival (PFS) for advanced lung squamous carcinoma patients who received chemotherapy. A total of 83 lung squamous carcinoma cases were divided into two groups by patients’ wishes. The control group contained 41 cases were treated by the chemotherapy of gemcitabine plus cisplatin (GP). The trial group contained 42 cases were treated by chemotherapy plus Chinese herbal medicine TBSS decoction. The toxicity and side effects of chemotherapy, as well as short-term outcome were evaluated. PFS of patient was recorded. The results showed that there were no differences on granulocytopenia (P = 0.115) or short-term outcome (P = 0.081) for patients of both groups after chemotherapy. The percentages of nausea, vomiting and thrombocytopenia in the trial group were lower than that in the control group (P = 0.037,P = 0.040). The PFS of patients in the trail group were prolonged compared to patients in the control group (4.31 ± 0.24 VS 3.78 ± 0.16 month;P = 0.043). It was concluded that Chinese herbal medicine TBSS decoction cannot reduce granulocytopenia caused by chemotherapy, or improve the tumor response rate (RR) of short-term outcome. However, it can prolong PFS, relieve nausea, vomiting and thrombocytopenia during chemotherapy.