Objective: To screen candidate antigen for therapeutic HBV vaccine with a novel adjuvant DC-Chol. Methods: BALB/c mice were injected with DC-Chol liposome and HBsAg prepared from CHO and Yeast respectively. One week later, IL-4, IL-2, IFN-?were measured by ELISA or ELISPOT. Results: The levels of IL-2, IFN-?of HBsAg from Yeast with DC-Chol liposome were 20 and 119 times higher respectively than those of HBsAg from CHO with DC-Chol liposome. ELISPOT assay showed that the counts of spot-forming cells of IL-4 and IFN-?of HBsAg from Yeast with DC-Chol liposome were 2.8 and 46.3 times higher respectively than those of HBsAg from Yeast with Al(OH)3. Conclusion: HBsAg prepared from Yeast together with DC-Chol liposome may be an appropriate candidate for therapeutic HBV vaccine .

OBJECTIVETo investigate the relationship between recurrent spontaneous abortion (RSA) and the level of interferon-γ (IFN-γ) and interleukin-4 (IL-4) in peripheral blood and gingival crevicular fluid of patients with chronic periodontitis.

METHODSFifty pregnant women with recurrent spontaneous abortion (gestational age: 5 - 28 weeks) and 40 pregnant women (gestational age: 39 - 41 weeks) were included and their periodontal status was examined. The level of IFN-γ and IL-4 in gingival crevicular fluid and peripheral blood was detected by enzyme-linked immunosorbent assay.

RESULTSNine RSA women and 5 in control group had periodontitis. The prevalence of periodontitis in RSA group was 18% (9/50) and 13% (5/40) in the control group (P < 0.05). The probing depth, attachment loss and bleeding index in RSA group were higher than those in the control group (P < 0.05). The level of IFN-γ was (52.98 ± 17.56) ng/L in gingival crevicular fluid of RSA group and (25.25 ± 7.93) ng/L in control group (P < 0.01). The level of IL-4 was (15.43 ± 1.77) ng/L in gingival crevicular fluid of RSA group and (19.62 ± 4.04) ng/L in control group (P < 0.01). The content of IFN-γ was (27.79 ± 3.59) ng/L in peripheral blood of RSA group and (18.39 ± 2.65) ng/L in control group (P < 0.05). The content of IL-4 was (15.88 ± 0.95) ng/L in peripheral blood of RSA group and (22.98 ± 4.30) ng/L in control group (P < 0.001). The content of cytokines in gingival crevicular fluid and peripheral blood in the same group was positively related (r > 0.8, P < 0.001).

CONCLUSIONSPregnant women of RSA have a higher chance of getting periodontitis. The change in the level of cytokines in periodontal tissue could result in excursion of maternal intrauterine immune environment to T-helper 1 (Th1). Periodontitis was related to RSA.

Abortion, Habitual ; blood ; metabolism ; Adult ; Chronic Periodontitis ; blood ; epidemiology ; metabolism ; Female ; Gingival Crevicular Fluid ; metabolism ; Humans ; Interferon-gamma ; blood ; metabolism ; Interleukin-4 ; blood ; metabolism ; Pregnancy ; Pregnancy Trimester, First ; Pregnancy Trimester, Second ; Prospective Studies

To explore the treatment of refractory anemia (RA), 7 cases of myelodysplastic syndrome (subtype of refractory anemia) were treated in combination of cyclosporin A (CsA) with stanozolol. Duration of treatment with CsA was 5 months-3 years (mean 13 months). The results showed that among 7 cases 6 were effective, 1 case no responded to treatment. 3 cases out of 6 effective cases achieved complete remission without transfusion dependence, 1 cases achieved partial remission, 2 cases were improved. During the investigation signs of leukemia ot other malignant tumors not were found in all cases. In conclusion, CsA treatment is effective for part cases of RA, side effects of drugs are tolerable for patients.
Adolescent ; Adult ; Aged ; Anemia, Refractory ; drug therapy ; Cyclosporine ; administration & dosage ; Drug Therapy, Combination ; Female ; Humans ; Male ; Middle Aged ; Stanozolol ; administration & dosage

OBJECTIVETo construct a high performance open source software engine based on IBM SPLASH algorithm for later research on pattern discovery.

METHODSGpat, which is based on SPLASH algorithm, was developed by using open source software.

RESULTSGNU Pattern (Gpat) software was developped, which efficiently implemented the core part of SPLASH algorithm. Full source code of Gpat was also available for other researchers to modify the program under the GNU license.

CONCLUSIONGpat is a successful implementation of SPLASH algorithm and can be used as a basic framework for later research on pattern recognition in biological sequences.

Algorithms ; Computational Biology ; Pattern Recognition, Automated ; Sequence Analysis, DNA ; Software

Objective:To stablish of chemiluminescent Southern blot detection system for examining HBV DNA replication intermediates in HepG2.2.15,and analyse the inhibition of HBV replication with three kind of drugs with different targets.Methods:The HBV DNA replication intermediates were extracted and analyzed by Southern blot with HBV probe,which(pTHBV1047) was labelling with digoxigenin.The results of the hybridization were detected by chemiluminescent,and the condition of hybridization was optimized.After treated with lamivudine,Bay41-4109,?-Galcer in different concentration,the HBV DNA from the HepG2215 cells were detected with the system.Results:the sensitivity of the system was 1pg of pTHBV1047,and HBV specific positive signals was detected with the DNA from HepG2.2.15.The three kinds of drugs can inhibit the HBV replication obviously with chemiluminescent Southern blot detection system,the IC50 were 1.53?mol/L,0.41?mol/L,0.01?mol/L.Conclusion:The HBV replication intermediates from the cell of HepG2.2.15 can reflect the antiviral effect accurately with different targets drugs and this mothod would be used in the study of Chinese-midicine.

A new species of genus shewarella Shewanellade decolorations S12, was isolated from activated sludge of a textile-printing waste-water treatment plant. In the anaerobic condition, S12 could conserve energy for growth by using Fe3 + as the terminal electron acceptor. At the optimal condition of pH8, temperature 30℃, ferric citrate 800mg/L, sodium lactate 2g/ L, yeast extract 0. 5g/ L , the cell growth increased with the raise of the amount of the Fe3+ reduction in 8k The effect of different carbon soucres, nitrogen sources, pH values and growth temperatures on the anaerobic Fe3 + reduction of Shewanella decolorations S12 was investigated. LB was favorable for Fe3 + reduction. Glucose and sodium lactate also were favorable for Fe3+ reduction. The cell growth and Fe3 + reduction increased with the raise of the amount of the yeast extract from 0 to 4g/L The amounts of the sodium lactate of 6g/ L and ferric citrate of 800mg/L were suitable for strain S12 growth and Fe3+ reduction. In the optimum initial pH value range of 6 -8 for Fe3+ reduction, strain S12 growth increased with the raise of the pH val- ue. Strain S12 could growth and reduce Fe3+ at the temperature range of 20 -40℃. The best temperature for strain S12 growth and Fe3 + reduction was 301.

Objective To detect plasma interleukins-35 (IL-35 )level in the patients with active tuberculosis complicated with bronchiectasis and to analyze its clinical significance.Methods Peripheral blood of patients with active tuberculosis from depart-ment of Dongguan 6th People′s hospital were collected,assigned to the active tuberculosis complicated with bronchiectasis group and active tuberculosis group.The healthy volunteers served as the control group.The plasma IL-35 level was measured by ELISA, and peripheral blood neutrophils and lymphocytes were detected by hematology analyzer.Results The levels of plasma IL-35 signif-icantly increased in both patients with active tuberculosis complicated with bronchiectasis and patients with active tuberculosis.The level of plasma IL-35 of patients with active tuberculosis complicated with bronchiectasis was significantly higher than that of the patients with active tuberculosis.The absolute value and percentage of peripheral blood neutrophils of patients with active tubercu-losis complicated with bronchiectasis were significantly higher than those of healthy volunteers.However,the percentage of periph-eral blood lymphocytes of patients with active tuberculosis complicated with bronchiectasis was significantly lower than that of healthy volunteers.Pearson correlation analysis showed that the absolute value of peripheral blood neutrophils of patients with ac-tive tuberculosis was positively correlated to the level of plasma IL-35.Conclusion IL-35 may play an important role in the progres-sion of active tuberculosis complicated with bronchiectasis.The determination of IL-35 may be helpful to the diagnosis of patients with active tuberculosis complicated with bronchiectasis.

Objective To knockout interferon alpha/beta receptor subunit 1(IFNAR1) gene in human colorectal adenocarcinoma cells Caco-2 using clustered regularly interspaced short palinmic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system to construct IFNAR1 knockout Caco-2 cell line.Methods The single guide RNA(sgRNA)sequence was designed to specifically recognize the exon region of IFNAR1 gene using CRISPR/Cas9 technology,and the LentiCRISPRv2-IFNAR1-sgRNA recombinant plasmid was constructed.Caco-2 cells were infected with the plasmid packaged by lentivirus and screened by puromycin resistance.The obtained monoclonal cell lines were cultured by limited dilution method,which were verified for the effect of IFNAR1 gene knockout by target gene sequencing and Western blot,and detected for the mRNA levels of CXC chemokine ligand 10(CXCL10)and interferon-stimulatd gene 20(ISG20)in IFNAR1knockout cells by adding exogenous IFNβ.Results Sequencing results of plasmid LentiCRISPRv2-IFNAR1-sgRNA showed that the insertion sites were all located at the sticky end of BsmBⅠenzyme digestion.Two IFNAR1 knockout monoclonal cell lines were obtained.The sequencing results showed that Caco-2-IFNAR1-KO1 had 5 bp deletion in the sixth exon of IFNAR1,and Caco-2-IFNAR1-KO2 had 18 bp deletion and 1 bp insertion in the seventh exon.Compared with wild-type Caco-2 cells,Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells showed no expression of IFNAR1 protein.Compared with no IFNβ stimulation,the mRNA levels of CXCL10 gene(t = 0.566 and 1.268 respectively,P>0.05)and ISG20 gene(t =1.522 and 1.733 respectively,P>0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells stimulated by 50 ng/mL IFNβ showed no significant increase.While compared with those of wild-type Caco-2 cells,the mRNA levels of CXCL10gene(t = 6.763 and 6.777 respectively,P<0.05)and ISG20 gene(t = 5.664 and 5.65 respectively,P<0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells decreased significantly under the stimulation of 50 ng/mL exogenous IFNβ.Conclusion Caco-2 cell line with IFNAR1 knockout was successfully constructed by using CRISPR/Cas9 technology,and the downstream molecules activated by IFNAR(interferon alpha/beta receptor)in this cell line were obviously inhibited,which provided a powerful tool for further exploration of the innate immune response and replication packaging mechanism of Caco-2 cells after virus infection.

Seven compounds were isolated from fermentation extract of cave-derived Metarhizium anisopliae NHC-M3-2 by silica gel, semi-preparative HPLC and other chromatographic methods. Their structures were elucidated by UV, IR, MS and NMR methods as 2,3-dehydroindigotide G (1), (-)-regiolone (2), naphtho-γ-pyrone (3), indigotide G (4), indigotide B (5) destruxin A (6) and destruxin B (7). Compound 1 is a new glycoside naphthopyranone compound. The anti-hepatitis B virus (HBV) activity of these compounds was evaluated. The EC₅₀ and CC₅₀ of compound 3 against HBV were 4.5 μmol·L^-1 and 92.3 μmol·L^-1, respectively. This is the first report of the antiviral activity of compound 3.

OBJECTIVEThis study aimed to explore the scene-trait coping style of military rescuers in Wenchuan earthquake in an effort to provide scientific evidences for mental intervention program for Chinese military personnel.

METHODSBy cluster sampling, a total of 151 military rescuers and 331 control servicemen were administered the military personnel scene-trait coping style scale (MPSTCSS).

RESULTSAll active coping factor scores, and passive coping factor scores of affection, health and economy in the rescuer group were significantly higher than those in control group (P < 0.05). The 21-above age subgroup, the 3-year plus service subgroup, and the officer subgroup had significantly higher active coping factor scores on military tasks, military experience and personal development than those of the 21-below age subgroup, 3-year minus service subgroup and the soldier subgroup, respectively (P < 0.05 or P < 0.01).

CONCLUSIONThe earthquake relief servicemen can cope with stressful situations better than control group by taking active coping style. The officers, servicemen older than 21 years, and servicemen with more service duration than 3 years could usually take active coping style.

Adaptation, Psychological ; Disasters ; Earthquakes ; Humans ; Military Personnel ; psychology ; Stress, Psychological