BACKGROUNDTo construct recombinant retroviral vector expressing HBcAg in eukaryotic cells.

METHODSThe HBV core gene fragment was amplified by using PCR from pADR which contains complement nucleotide sequence of HBV subtype adr and cloned into retroviral expression plasmid pLXSN, then transfected into packing cell (PT67) with lipofec AMINE. After 2-3 weeks selection with G418, large colonies were isolated and transferred to individual plates. Virus-containing medium was collected and used to infect NIH3T3, EL4 and mouse bone marrow derived dendritic cells(DC). DNA was extracted from infected cells and tested by PCR. Indirect immunofluorescence and FACS were used to detect the expression of HBcAg. Cell mediated immunity of immunized C57BL/6 mice with transduced DC was analyzed.

RESULTSThe insertion of HBV core gene fragment in the recombinant retroviral plasmid was confirmed by PCR as well as enzyme digestion with EcoR1 and BamH2. The viral titer reached 3 x 10(5) CFU/ml. The result of PCR showed that the HBV core gene had been integrated into the genome of infected NIH3T3 cells. Indirect immunofluorescence and FACS analysis showed that HBcAg had been expressed in these cells. HBcAg specific CTL responses could be generated in mice immunized with retrovirus transduced DC.

CONCLUSIONSHBV core gene had been integrated into eukaryotic cells with retroviral vector and target gene had been expressed efficiently. These results may have some impact on gene therapy of chronic hepatitis B.

3T3 Cells ; Animals ; Cloning, Molecular ; Dendritic Cells ; metabolism ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; genetics ; Humans ; Mice ; Recombination, Genetic ; genetics ; Retroviridae ; genetics ; Transfection

OBJECTIVETo investigate the factors of long-turn survival of liver cancer after surgical treatment.

METHODSFive hundred and twenty-two cases of liver cancer that received surgical treatment in 14 years were analyzed retrospectively.

RESULTSComparison between the small liver cancer (< 5 cm) and the greater one (> 10 cm) revealed that the small liver cancer had a higher survival rates than the greater one [3 year (61.25 +/- 4.41)% versus (45.90 +/- 6.98)%; 5 year (53.84 +/- 5.68)% versus (30.21 +/- 10.23)%]. There were same results between single-nodule and two or more than two nodule [3 year (61.86 +/- 3.69)% versus (38.31 +/- 4.97)%; 5 year (55.40 +/- 4.91)% versus (28.01 +/- 6.31)%], between child I and child II or more than II [3 year (60.68 +/- 3.68)% versus (49.88 +/- 4.13)%; 5 year (50.99 +/- 5.10)% versus (36.39 +/- 7.58)%], and between single segmentectomy of the liver and two or more than two segmentectomy [3 year (68.65 +/- 4.95)% versus (49.88 +/- 4.13)%; 5 year (65.38 +/- 5.69)% versus (37.98 +/- 5.70)%].

CONCLUSIONSSmall liver cancer, single-nodule, good hepatic function and minor resection were important factors to prolong survival further.

Adolescent ; Adult ; Aged ; Child ; Female ; Follow-Up Studies ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; mortality ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate

The purpose of the present study was to examine the kinetic process of hemoglobin (Hb) carrying and releasing oxygen. Under the standard conditions (pH 7.4, Po(2) 20 mmHg, 20 °C) the blood samples of chicken, rabbit, frog and carp were equilibrated in oxygen content analyzer with calibrated gas mixture A (0.5% CO(2) and 99.5% N(2)). Then the blood samples were exposed to gas mixture B (21% O(2), 0.5% CO(2) and 78.5% N(2)). After equilibration, the blood samples were exposed to gas mixture A again. During the whole process, Po(2) of blood samples was detected in real-time. The time spent in blood Po(2) changing from 0 to 21 kPa was recorded carefully. The results indicated that the kinetic curve of Hb carrying oxygen presented a shape of "S". It was similar to the Hb oxygen dissociation curve (Hb ODC). Based on the curve, T(50), a new kinetic parameter, was established. T(50) is the time of 50% O(2) saturation of Hb. It can reflect the efficiency of Hb carrying oxygen. Through comparing of T(50), the efficiency of Hb carrying oxygen among 4 species of animals was: frog < carp < rabbit < chicken. In the phase I of Hb carrying and releasing oxygen kinetic curve, the slope in carp was much larger than that in rabbit; the time [(1 411±6) s] of Hb releasing oxygen in chicken was longer than that in other 3 animals. These differences reflected the variety of efficiency of Hb carrying and releasing oxygen. In addition, the kinetic features of Hb carrying oxygen were likely to become an important index to evaluate the function of Hb carrying oxygen, especially in evaluating the ability of artificial blood substitute. On the basis of the analysis of the kinetic curve of Hb carrying oxygen and Hb ODC, another new important efficacy parameter E(50) was proposed. E(50) reflects the relationship between the time of 50% O(2) saturation of Hb and environmental Po(2). E(50) can be used as a synthetic index to assess the efficiency of Hb carrying oxygen.
Animals ; Anura ; Carps ; Chickens ; Hemoglobins ; metabolism ; Hydrogen-Ion Concentration ; Kinetics ; Oxygen ; metabolism ; Rabbits

Objective To investigate selcnium(Se) levels of environment and human body in Gaomi City and Zichuan District of Shandong. Methods Lijiaying Township in Gaomi City of Weifang City, Zhaili Township and Longquan Township in Zichuan District of Zibo City were selected. Two farming soil samples at different spot, local wheat and corn, residents nail samples from 3 to 4 families were collected in each natural village in the investigated towns. The contents of Se were detected by 2,3-diamino naphthalene fluorescence method. Results Se level of the soil, wheat, corn, and nails in Lijiaying [(0.054 ± 0.019), (0.022 ± 0.009), (0.018 ± 0.007), (0.365 ± 0.108)mg/kg] was significantly lower than that in Zhaili [(0.425 ± 0.080), (0.130 ± 0.043), (0.098 ± 0.026), (0.751 ± 0.134)mg/kg] and Longquan[(0.487 ± 0.153), (0.112 ± 0.030), (0.097 ± 0.029), (0.735 ± 0.145)mg/kg;P < 0.01]. In Lijiaying, Se was deficient in soil, wheat, corn(< 0.200, < 0.025 mg/kg), above Se deficiency diagnosis and below Se-adequate level in the nail, while in Zhaili and Longquan, the Se level in the soil (0.425, 0.487 mg/kg), wheat(0.130, 0.112 mg/kg), corn (0.098, 0.097 mg/kg), nails (0.751, 0.735 mg/kg) was adequate (≥0.400 mg/kg). Conclusions The external environment is Se-deficient in Lijiaying, Se-adequate in Longquan and Zhaili. The selenium level in human body is consistent with the external environment.

OBJECTIVETo investigate the clinical effect of atlantoaxial pedicle screw internal fixation for treatment of atlantoaxial dislocation.

METHODSSixteen patients with atlantoaxial dislocation were treated from Dec. 2005 to June 2007, included 10 males and 6 females, aged from 38 to 45 years old (means 40.5 years). Among them 12 patients combined with nerve injury, according to ASIA grade: there were 3 cases in grade B, 5 cases in grade C, 4 cases in grade D. All patients received preoperative CT, radiograph and skull traction. Intraoperative posterior approach general spine pedicle screw-rod orthopaedics fixation system used and iliac bone block were implanted in space of posterior atlantal arch and axial vertebral plate. The outcome and complications were observed in the near future.

RESULTSThere was no vascular or neural injury found. The patients were followed up for 12 to 24 months (means 18 months). All head pain, acid storm symptoms were improved after operation. According to the Odom's clinical efficacy evaluation standard, 12 cases were excellent, 4 were good. Eleven cases of 12 with nerve injury recovered significantly. By ASIA classification: 1 cases was in grade B, 2 cases were in grade C, 5 cases were in grade D, 4 cases were in grade E. No looseness or breakage of screw occurred. Bony fusion was achieved in all cases.

CONCLUSIONPosterior approach atlantoaxial pedicle screw internal fixation have the advantages of direct screw placement, short-segment fusion, intraoperative reduction, fixation reliable, high fusion rate, and it can restablish the upper cervical vertebrae stability and help to recover the spinal cord and nerve function.

Adult ; Atlanto-Axial Joint ; diagnostic imaging ; injuries ; physiopathology ; surgery ; Bone Screws ; Female ; Fracture Fixation, Internal ; instrumentation ; Humans ; Joint Dislocations ; diagnostic imaging ; physiopathology ; surgery ; therapy ; Male ; Middle Aged ; Tomography, X-Ray Computed ; Treatment Outcome

The free membrane of Eudragit L100/S100 which is pH-sensitive, colon-specific was prepared by plane casting films. The film humidity, species and amount of plasticizers, the ratio of membrane material was investigated. The rate of membrane permeability and mechanical properties were used as indicators of orthogonal experiment, and its related properties were studied. The results show that the mechanical properties of the membrane and phragmoid capacity are the best when 30% TEC was used as plasticizer; the ratio of membrane material have little effect on the rate of membrane permeability and mechanical properties. By adjusting the species and amount of plasticizers, the ratio of Eudragit L100/S100, the free membrane which is colon-specific can be obtained.
Citrates ; chemistry ; Humidity ; Hydrogen-Ion Concentration ; Permeability ; Phthalic Acids ; chemistry ; Plasticizers ; chemistry ; Polymethacrylic Acids ; chemistry ; Technology, Pharmaceutical ; methods ; Tensile Strength ; Transition Temperature ; Triacetin ; chemistry

OBJECTIVETo investigate methods and therapeutic effects of sequential drugs treatment for central pain following spinal cord injury.

METHODSA total of 28 patients suffered from central pain following spinal cord injury were treated with sequential drugs from 1994 to 2008, including 23 males and 5 females, ranging in age from 25 to 59 years (mean 42 years). According to the patients' response to drugs, the therapy grade was adjusted step by step until the pain was relieved. Basing on VAS scores before and after drugs treatment, analgesic effect was evaluated. The first grade drugs: COX-2 inhibitors. The second grade drugs: Tricyclic antidepressant drugs (Amitriptyline) + COX-2 inhibitors + Carbamazepine. The third grade drugs: Tricyclic antidepressant drugs (Amitriptyline) + Gabapentin + Neurotropin/COX-2 inhibitors.

RESULTSThe pain of all of 28 patients was relieved to different extent. The VAS scores decreased by 23.3 +/- 1.2 in the first grade drugs treatment group. The VAS scores decreased by 54.5 +/- 3.8 in the second grade drugs treatment group. The VAS scores decreased by 65.8 +/- 5.1 in the third grade drugs treatment group (P<0.05).

CONCLUSIONThe sequential drugs treatment for central pain following spinal cord injury has a good analgesia effect and little adverse reaction.

Adult ; Amitriptyline ; administration & dosage ; Carbamazepine ; administration & dosage ; Cyclooxygenase 2 Inhibitors ; administration & dosage ; Drug Therapy, Combination ; Female ; Humans ; Male ; Middle Aged ; Pain ; drug therapy ; Pain Measurement ; Polysaccharides ; administration & dosage ; Spinal Cord Injuries ; complications ; drug therapy

OBJECTIVETo investigate the specificity and sensitivity of the immunohistochemistry for hMLH1 and hMSH2 with detection of microsatellite instability (MSI) to identify the kindreds with hereditary nonpolyposis colorectal cancer and to analyse its value in clinical practice.

METHODSpecimens of 16 cases with HNPCC and 16 cases with sporadic colorectal cancer were detected by immunostaining with hMLH1 and hMSH2 and MSI was also detected.

RESULTSThe specificity and sensitivity of the immunohistochemistry for hMLH1 and hMSH2 were 91.7% and 87.5% respectively. The specificity and sensitivity of MSI were 100% and 75.0%. By combining two methods, the specificity and sensitivity were 91.7% and 93.8% respectively.

CONCLUSIONSBy combination of the immunohistochemistry for hMLH1 and hMSH2 and detection of MSI to identify the kindreds with HNPCC, the specificity and sensitivity are improved which is better than to use either of them alone. And it is very easy and cheap that it can be used in clinics.

Adaptor Proteins, Signal Transducing ; Adult ; Aged ; Carrier Proteins ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; DNA-Binding Proteins ; analysis ; Female ; Genomic Instability ; Humans ; Immunohistochemistry ; Male ; Microsatellite Repeats ; Middle Aged ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; Neoplasm Proteins ; analysis ; Nuclear Proteins ; Proto-Oncogene Proteins ; analysis ; Sensitivity and Specificity

Objective :To explore influence of dual-and triple-anticoagulant therapy on hemorrhage events ,laboratory in-dexes and cardiovascular events after percutaneous coronary intervention (PCI) in patients with atrial fibrillation (AF) complicated coronary heart disease (CHD).Methods : Clinical data of 100 AF + CHD patients were analyzed .All subjects received PCI and anticoagulant therapy after PCI .Dual-therapy group (n＝ 50) received dual-anticoagulant program and triple-therapy group (n＝50) received triple-anticoagulant program.Levels of blood C reactive protein (CRP) ,fibrinogen (Fg) ,D-dimer (D-D) and platelet count (PLT) before and after treatment ,incidence of hemorrhage events and adverse cardiovascular events during treatment were compared between two groups .Results : Compared with dual-therapy group af-ter 12-month treatment ,there were significant reductions in blood levels of CRP [(16-81 ± 4-49) mg／L vs.(7-86 ± 1-64) mg／L] ,PLT [ (298-47 ± 54-33)×109／L vs.(148-89 ± 23-11)×109／L] ,Fg [ (5-59 ± 1-41) g／L vs.(2-14 ± 0-56) g／L] and D-D [(0-37 ± 0-08) mg／L vs.(0-21 ± 0-06) mg／L] in triple-therapy group ,P＝0-001 all.Incidence rate of ad-verse cardiovascular events in triple-therapy group was significantly lower than that of dual-therapy group (6-00% vs. 22-00%, P＝0-021) ,there was no significant in incidence rate of hemorrhage events between two groups , P＝0-182-Conclusion : Triple-anticoagulant therapy can significantly improve therapeutic effect with high safety in AF + CHD pa-tients after PCI ,which is worth extending .

Objective To determine the carriage prevalence and antibiotics resistance profile of methicillin-resistant staphylococcus aureus(MRSA) and methicillin-sensitive staphylococcus aureus(MSSA) strains isolated from newborns. Methods A cross-sectional study was conducted in two hospitals in Shenzhen from August to November 2015. Cotton swab samples were collected from 1834 newborns. Staphylococcus aureus was identified by general laboratory tests, and antimicrobial-susceptibility profiles was tested. Results In total, 3.27% (60/1 834) Staphylococcus aureus (SA) strains were isolated, including 0.82% (15/1 834) MRSA and 2.45% (45/1 834) MSSA. Most MRSA and MSSA strains were resistant to penicillin and erythromycin but less than 10% strains resistant to linezolid. Drugs resistance rates of MRSA to erythromycin, moxifloxacin and clindamycin were all higher than that of MSSA (all P<0.05). The multidrug-resistant risk of MRSA (80.00%) was 12.36 times higher than that of MSSA (24.44%). Conclusions Multidrug-resistant SA especially MRSA strains isolated from newborns had the higher prevalence, the drugs resistance rate of MRSA strains was more serious than that of MSSA.