AIM:To observe the preliminary efficacy of conbercept injected intravitreally for the treatment of wet age-related macular degeneration(wAMD).METHODS:Seventeen wAMD patients (18 eyes) were selected to receive conbercept injection.All patients were given a single conbercept injection every month,3 times.Before and after 1,2,3mo of the injection,the best corrected visual acuity (BCVA),intraocular pressure (IOP,measured by Non-contact tonometer),fundus photography,fundus fluorescein angiography(FFA),indocyanine green angiography(ICG),optical coherence tomography(OCT) examination and the complications incidence were compared.RESULTS:Three months after conbercept injection,the BCVA improved in 15 eyes (83%),stable in 3 eyes (17%).Before treatment,the average central macular thickness was 421.72±54.43μm,at 1 and 2 and 3mo after treatment,the average central macular thickness was 337.89±25.88μm,293.56±26.87μm,266.89±19.10μm respectively.There were significant differences compared with before and after injection(P<0.05).In the final follow up,FFA and ICG showed that the leakage in macular area disappeared in 15 eyes (83%),still existed in 3 eyes (17%),in those 3 eyes the injection was given for one or two times till the leakage disappeared.Elevated intraocular pressure occurred in 2 cases (26mmHg,23mmHg),after 1d down to normal.Another patient showed postoperative envy,given left ofloxacin eye drops after 2d,then back to normal.There was no serious ocular adverse reactions.CONCLUSION:Intravitreal injection conbercept for wAMD can significantly improve the visual function,reduce the macular edema and the leakage with higher safety and less complications.However the prolonged efficacy needs further observation.

Acute Disease ; Adult ; Chromatography, Gas ; Female ; Humans ; Lung ; chemistry ; Phosphines ; analysis ; blood ; poisoning

Objective To assess the diagnostic value of MRCP before LC.Methods 944 cases with chronic calculous cholecystitis underwent MRCP before LC from June 2004 to June 2007 in our department.incidence rate of cholecvstolithiasis together with common bile duct stones and incidence rate of anatomic abnormity of bile duct were collected.Results The incidence rate of cholecvstolithiasis together with common bile duct stones were 8.1%(77/944),and the oecurence ofACBDS were 1.2%(11/944).The incidence rate of anatomic abnormity of bile duct were 3.7%(35/944).ConclusionMRCP can not only offer a excellent diagnostic value of ACBDS and anatomic abnormity of bile duct,but also reduce the occurrence of CBDS remainder and iatrogenic bile duct iniuries.

Objective To establish a big animal model of secondary infection of acute necrotizing pancreatitis (ANP). Methods Thirty young pigs were allocated to experiment group ( n = 20 ) or control group (n = 10). The ANP model was induced by retrograde injection of a mixture solution of 5% sodium taurocholate and 5% trypsin (0. 5 ml/kg body weight) into the main pancreatic duct and ligation of the proximal end of the main pancreatic duct, and then the second step was injecting 3 ～ 4 ml living Escherichia coli (E coli) suspension (108/ml) to the necrotic area of the pancreas by fine needle aspiration technique under CT guidance in the experiment group, and by injecting 3 ～ 4 ml inactivated E coli in the control group using the same method. Multi-slice spiral CT dynamic enhanced scan was performed in both groups 1 day and 2 or 3 days after ANP modeling and 5 days after bacterial injection to calculate the CTSI score. Serum amylase, blood WBC count and blood bacterial culture was performed in both groups. 5 days later, the animals were scarified to observe the infected or necrosis foci, and perform smear, bacterial culture and pathologic examinations of the tissue around the infected or necrosis foci. Results The ANP secondary infection model was successfully established in 16 of the 20 animals in the study group, with a success rate of the 80.0% (16/20). There were 17 foci where the positive rate of bacterial culture was 100% (17/17 foci), and the success rate of blood bacterial culture was 68.8%(11/16). In the control group, the ANP model was established successfully in 7 of 10 animals (70%), except for one case of contamination, only one foci was identified;the positive rate of bacterial culture and the success rate of b|ood bacterial culture was 14.3% (1/7). Serum amylase and white blood WBC count increased with similar trends, WBC count in the study group was significantly higher than that in the control group (P<0.01). The mean CT severity index(CTSI) was all ≥4 in beth groups, indicating the severity was moderate to severe. Conclusions A stable and reliable model of secondary infection of ANP in big could be established satisfactorily by injecting active E. coli into the pancreatic necrosis tissue under CT guidance, which helps further pathogenic mechanism studies and clinical studies, especially imaging studies.

Objective To explore the mechanism of protecting cells from hypoxia/ reoxygenation (H/R) injury by constructing specific small interference RNA (siRNA) to inhibit NALP3 expression in rat renal tubular epithelial cells (NRK-52E).Methods (1) To establish the H/R injury model of NRK-52E by regulating the pressure of N2 in incubator to hypoxia condition,the cells were cultured with hypoxia for 1 h and then with reoxygenation for 1 h,2 h,4 h,8 h,16 h and 24 h.The activity of lactae dehydrogenase (LDH) in the culture medium,cell count and cell viability,the expression of NALP3 were determined by biochemical method,trypan blue exclusion and Western blotting.(2) The siRNA was transfected into NRK-52E.The irrespective siRNA transfected group wasused as control.NALP3 expression was examined by Western blotting.(3) The cells were divided into 4 groups:control group,H/R group,irrespective siRNA transfected group and NALP3-siRNA transfected group.To establish the H/R injury model of NRK-52E by regulating the pressure of N2 in incubator to hypoxia condition,the cells were cultured with hypoxia for 1 h and then with reoxygenation for 4 h.And the expression of NALP3 was determined by Western blotting.(4)Cellular apoptosis was examined by Annexin V/PI staining and flow cytometry.NF-κB DNA binding activity,IκB-α,Bcl-2 and Bax expression were examined by EMSA and Western blotting.Results (1)Compared with the control group,the activity of LDH significantly increased,cell count and cell viability significantly decreased (all P＜0.05).The expression of NALP3 significantly increased and peaked at 4 h after H/R.(2)The specific siRNA could efficiently inhibit NALP3 expression in NRK-52E.Compared with the irrespective siRNA transfected group,the protein expression of NALP3 was significantly down-regulated in NALP3 siRNA transfected group (P＜0.05).(3)After hypoxia 1 h and reoxygenation 4 h,the activity of LDH and the expression of NALP3 increased.Compared with the irrespective siRNA transfected group,LDH concentration in media and the expression of NALP3 significantly decreased in NALP3-siRNA transfected group.(4)After hypoxia 1 h and reoxygenation 4 h,NF-κB DNA binding activity was increased,IκB-α phosphorylation and degradation,Bcl-2 and Bax were significantly up-regulated.However,compared with the irrespective siRNA transfected group,NF -κB DNA binding activity,IκB-α degradation and Bax/Bcl-2 were significantly decreased (P＜0.05) in NALP3-siRNA transfected group.At the same time,the ratio of apoptosis was significantly increased in three groups than that in control.Compared with the irrespective siRNA transfected group,the ratio of apoptosis in NALP3-siRNA transfected group was significantly decreased (P＜0.05).Conclusions H/R induces the expression of NALP3 in NRK-52E.The synthesized siRNA can inhibit the expression of NALP3 and protect NRK-52E from hypoxia/reoxygenation injury.The mechanism may be via inhibiting the activation of NF-κB,modulating expression of Bcl-2 and Bax,as well as decreasing cell apoptosis.

OBJECTIVETo detect the binding sites and characteristics of the adduct from the reaction of formaldehyde, acetaldehyde, acrolein with DNA.

METHODSFormaldehyde, acetaldehyde, acrolein were reacted with four kinds of deoxyribonucleoside monophosphate (dNMP) in buffered solutions with neutral pH. The reaction products were separated by high performance liquid chromatograph (HPLC) and characterized by UV spectroscopy.

RESULTSThe reaction of formaldehyde, acetaldehyde with dG was separated and detected by HPLC. The reaction of acrolein, formate, acetic acid, Mercapturic acid with dG was not separated and detected by HPLC, while the dominant dNMP binding with formaldehyde, acetaldehyde was also determined.

CONCLUSIONFormaldehyde, acetaldehyde could bind with dGMP to express genotoxic effects.

Acetaldehyde ; chemistry ; toxicity ; Acrolein ; chemistry ; toxicity ; Aldehydes ; chemistry ; toxicity ; Chromatography, High Pressure Liquid ; DNA ; chemistry ; DNA Damage ; drug effects ; Formaldehyde ; chemistry ; toxicity ; Guanosine Monophosphate ; chemistry

Objective:To investigate the efficiency of carbon nanotube(CNT)-PAMAM mediated entrance of anti-survivin oligonucleotide into HepG2 cells,and its effects on the proliferation of HepG2 cells.Methods:CNT-PAMAM-anti-survivin oligonucleotide compounds were prepared and characterized by AFM and 1% agarose gel electrophoresis analysis.TEM was used to observe the distribution of CNT-PAMAM-ASODN compounds in HepG2 cells.CNT-PAMAM-ASODN compounds were added into the medium and co-cultured with HepG2 cells for 24 h,48 h,72 h,and 96 h at 37℃,5% CO_2.MTT method was used to detect the effects of ASODN and CNT-PAMAM-ASODN on the proliferation of HepG2 cells.Results:CNT-PAMAM-ASODN compounds were successfully synthesized via AFM and agarose gel electrophoresis.TEM showed that the compounds were located in the cytoplasm.When CNT-PAMAM-ASODN(1.0 ?mol/L)and ASODN(1.0 ?mol/L)were used for a 48 h culture,the inhibitory rates of HepG2 cells were(45.97?4.28)% for CNT-PAMAM-ASODN compounds group,(9.33?0.85)% for ASODN group,and(6.37?0.69)% for CNT-PAMAM group.CNT-PAMAM-ASODN compounds at 1.5 ?mol/L inhibited HepG2 cells by(70.22?7.25)%,and the inhibitory effects were in a time-and concentration-dependent manner.There was statistical difference between experiment group and control group(P

OBJECTIVE To investigate the HIV infections states in tumor patients for clinical diagnosis,treatment and to prevent HIV infection in the tumor hospital.METHODS The result of HIV detection in tumor patients from Dec 2000 to Aug 2006 was analyzed by the review statistics analysis.RESULTS Totally 48 101 tumor paients were detected,and the number of tumor patients with positive HIV antibody result was 51(0.106%).Among the positive patients there were 21 cases with blood transfusion history,14 cases with blood donating experience,2 cases with both these two kinds of experiences and 14 cases without the two kinds of experiences.Their rate was separately 41.0%,27.5%,4.0% and 27.5%.Most of the HIV positive patients had no clinical synptoms.CONCLUSIONS The HIV positive rate of patients with blood transfusion or blood donating is significantly higher than the patients without these experiences.The routine detection for the HIV before the operation,blood transfusion or other traumatic detection is very necessary.

Objective To construct plasmid vectors of calmodulin(CaM)Mg2+binding site mutants,and to express,purify and identify the mutant proteins. Methods Three kinds of cDNAs coding for the mutated CaM were cloned into pGEX?6P?3 plasmid vectors. These recombinant plasmids were transfected into Escherichia coli BL21 to express GST fusion proteins of CaM mutants. The fusion proteins were purified with Glutathione?Sep?harose 4B beads and PreScission protease. Results Both enzyme digestion analysis and DNA sequence identification proved the successful con?struction of the CaM mutant plasmids. SDS?PAGE results showed the high purity of each CaM mutant protein. The concentrations of three CaM mu?tants were around 1.0 mg/mL. Conclusion Prokayotic expression vectors of CaM Mg2+binding site mutants were successfully developed,and the eli?gible CaM mutant proteins were obtained. This study provided an important basis for further study on CaM’s biological function.

Objective To investigate a method for the purification of the N?terminal peptide fragment(NT)of the myocardial calcium channel Cav1.2,and characterize its interaction with calmodulin(CaM). Methods EscherichiacoliBL?21 cells were transformed with plasmid pGEX?6p?3/NT harboring the NT?GST fusion gene. The cells harboring pGEX?6p?3/NT were cultured and protein expression was induced with isopropyl?β?D?thiogalactoside(IPTG). Then,the GST?NT fusion protein was purified by using glutathione Sepharose 4B(GS?4B)beads. GST was cleaved off with the PreScission protease,and SDS?PAGE was performed to detect the purity and relative molecular weight of the purified peptide. Further, GST pull?down assay was performed to characterize the interaction of the NT peptide with CaM. Results SDS?PAGE analysis showed that the NT peptide was successfully purified,with high purity. Results of the GST pull?down assay showed that the NT peptide could interact with CaM. Conclusion This study establishes a method for the purification of the NT peptide and lays the foundation for further research on the interaction partners and biological functions of NT.