Objective To construct and express a recombinant plasmid pGEX-Sj 14-3-3-Sj32 of Schistosoma japonicum in Escherichia coli (E.Coli) BL21 (DE3).Methods Sj14-3-3 and Sj32 antigen genes were amplified by PCR from template of plasmids pGEX-Sj14-3-3 and pET28α-Sj32 which were extracted from recombinant bacteria BL21 (pET28α-Sj32) and BL21 (pGEX-Sj14-3-3) stored in Institute of Infectious and Parasitic Disease of the First Affiliated Hospital of Chongqing Medical University.Sj14-3-3-Sj32 fusion gene obtained with gene SOEing was cloned into the vector pGEX-1λT to construct pGEX-Sj14-3-3-Sj32 which was identified by double digestion.The recombinant plasmid pGEX-Sj 14-3-3-Sj32 was transformed into E.Coli BL21 (DE3).The recombinant strains were induced by isopropyl-β-d-thiogalactoside (IPTG),and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.Results The 1 750 bp Sj14-3-3-Sj32 fusion gene was successfully amplified by gene SOEing and cloned into the vector pGEX-1 λT verified by restriction analysis,the recombinant plasmid pGEX-Sj 14-3-3-Sj32 was successfully constructed.The molecular mass of the expressed recombinant protein was proximately 73 × 103 as detected by SDS-PAGE.Western blotting confirmed that the expressed protein could be recognized by the immune sera from rabbit infected with Schistosomajaponicum.Conclusion The recombinant plasmid pGEX-Sj14-3-3-Sj32 is successfully constructed and could be highly expressed in E.coli and the expressed recombinant protein has specific antigenicity.

Objective To construct a recombinant Bifidobacterium bifidum (Bb)vaccine[Bb (pGEX-Sj26GST-Sj14-3-3)] of Schistosomajaponicum (Sj) and analyze the expression of the fusion gene Sj26GST-Sj14-3-3 of Sj in Bb.Methods The recombinant plasmid pGEX-Sj26GST-Sj14-3-3 was electroporated into Bb to construct a recombinant Bb (pGEX-Sj26GST-Sj14-3-3) vaccine.Mter induction with isopropyl-β-D-thiogalactoside (IPTG),double restriction enzymes digestion and polymerase chain reaction (PCR) were used to identify the recombinant Bb (pGEX-Sj26GST-Sj14-3-3),expression of the recombinant protein was analyzed and identified by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.Results The recombinant plasmid pGEX-Sj26GST-Sj14-3-3 was successfully transformed into Bb identified by double restriction enzymes digestion and PCR.SDS-PAGE analysis showed that the relative molecular mass of the expressed recombinant protein was approximately 67 × 103.The expressed protein could be recognized by the immune sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant Bb (pGEX-Sj26GST-Sj14-3-3) vaccine of Sj is successfully constructed.The fusion gene Sj26GST-Sj14-3-3 can be expressed in recombinant Bb and the expressed target protein shows specific antigenicity.

Objective To explore the dose rate response characterization of four kinds of dosimeters for clinical application.Methods Within the range of 100-600 cGy/min,the dose rate responses of the PTW 0.6 cm3 ion chamber,0.015 cm3 ion chamber,Matrixx Evolution 2D diode array and MapCHECK 2D diode array under the same measuring conditions were measured.The dose rate response of the PTW 0.6 cm3 ion chamber under different energy and working voltage were analyzed.Results All ionization chambe.r types of measured equipment showed certain dose rate dependence for 6 MV X-rays.All the differences were below 1％.The dose rate dependence disappeared for 15 MV X-rays.The 2D diode array had strong dose rate dependence and the response difference was about 2％.Conclusions It is necessary to test and analyze the dose rate response of the measured equipment in treatment technology with dose rate varying,in order to ensure the precision of daily calibration and dose verification.

Objective:To isolate endophytic fungi from Panax ginseng and study their antifungal and antitumor activities.Methods:Endophytic fungi were isolated from 5-year-old garden ginseng and 15-year-old transplanted ginseng.The antifungal active endophytes were screened with Pyricularia oryzae P-2b model using mircodilution method,and the activities of endophytes against pathogenic fungi were tested in vitro.The antitumor activities of the endophytes were examined by MTT method in vitro.Results:Sixteen(33.3%)of the 48 endophytic fungi fully suppressed the activity of P.oryzae P-2b;11(22.9%)colonies showed satisfactory antifungal activities against Candida albicans,Cryptococcus neoformans,Trichophyton rubrum,and Aspergillus fumigatus;and 5(10.4%)colonies showed satisfactory antitumor activities against tumor cell lines MKN45,LOVO,HepG2,and HL-60.Among the bioactive colonies,Yuan-25 showed best antifungal activity,with its MIC80 aganinst Trichophyton rubrum being 4 mg/L,which was similar to that of fluconazole.Yuan-27 showed the best antitumor activity,with its IC50 similar to that of doxorubicin.Conclusion:Isolated endophytic fungi of Panax ginseng has antifungal and antitumor activities and is worth further exploring.

Objective To clinically observe obstructive sleep apnea syndrome(OSAS) before and after cerebrovascular disease attacking and find prevention ways.Methods Seventy cases of berebrovascular patients were observed the OSAS status and the clinic data were collected in the mean time.Results We found 70% patients with snore and suffocation,89% with day drowse before attacking.After attacking,82% in-patient cases appeared different degree lower ventilation or apnea during 1～7d sleeping,which the average paroxysm frequency were 79 times per night and the average time were 72s per paroxysm.The therapeutic effects were good through different measures.Conclusion The active measures should be taken on OSAS after cerebrovascular disease attacking,which has a high frequency of OSAS.

Child ; Congresses as Topic ; Humans ; Pediatrics

Objective To investigate the distribution and antibiotic resistance of isolated pathogens in neonatal sepsis. Methods The results of blood culture and drug susceptibility test in neonates sepsis from January 2012 to June 2013 were retro-spectively analyzed. Results One hundred and thirty-two strains were detected in the blood samples, with 100(75.76%)Gram-positive bacteria, 30 (22.73%) Gram-negative bacteria and 2 (1.52%) fungus. Staphylococcus epidermidis, Escherichia coli and Staphylococcus aureus were the three most common pathogens. Gram-positive cocci was strongly resistant to penicillin (100.00%), erythromycin, selectrin and ampicillin/sulbactam (62.50%-100.00%), but still sensitive to vancomycin and teico-planin. The resistance rate of Gram-negative bacilli to ampicillin was 100.00%, and the resistance rate to cefatriaxone, selectrin and cefuroxime was 61.54%-100.00%. The resistance rate to imipenem and piperacillin/tazobactam was lower. Conclusions The selection of sensitive antibiotics should be based on the pathogens and drug resistance testing for the treatment of neonatal sepsis.

This paper reviewed the practice of Handicap International (HI) in China and discussed the HI localized action plan using current theories and approaches of community-based rehabilitation.

Abstract: AIDS combined with Pneumocystis jirovecii pneumonia (PJP) and disseminated infections of Talaromyces marneffei and Cryptococcus neoformans are rare. This paper summarizes and analyzes the diagnosis and treatment of an AIDS patient with multiple fungal infections for reference. A 79-year-old male patient was admitted to the hospital with "stool habit change for more than 20 days". The white blood cell count was 4.57×109/L, the percentage of neutrophils was 81.8%, the absolute count of CD4+ lymphocytes was 6/μL, and the CD4/CD8 ratio was 0.17. HIV antibody positive was confirmed by CDC. The cerebrospinal fluid and alveolar lavage fluid were positive for Cryptococcus neoformans capsular antigen, and Pneumocystis jirovecii was found by the bronchoalveolar lavage fluid stained with hexamine silver. The cerebrospinal fluid culture was positive for Cryptococcus neoformans, and the blood culture was positive for Cryptococcus neoformans and Talaromyces marneffei. CT showed that bronchovascular bundles in both lungs were more thick, patchy and cable-like high-density shadows were seen in both lungs, and the edges were blurred. Nodular and cable-like high-density shadows were seen in the posterior apical segment of the left upper lobe, with clear margins. Infection of both lungs was considered, and secondary pulmonary tuberculosis occurred in the left upper lobe. After admission, the patient was treated with various anti-bacterial and fungal drugs due to recurrent fever, but the effect was not effective. The fever symptoms of the patient could not be significantly improved, and his condition continued to worsen, and he eventually died. The patient with AIDS complicated with bacterial and fungal infection, especially PJP infection in serious condifiton and has a poor prognosis for rapid development, so clinical attention should be paid to. 

OBJECTIVETo investigate the clinical effect and safety of anti-D immunoglobulin (anti-D) in the treatment of children with newly diagnosed acute immune thrombocytopenia (ITP) through a Meta analysis.

METHODSPubMed, EMBASE, Cohrane Library, Ovid, CNKI, and Wanfang Data were searched for randomized controlled trials (RCTs) published up to April 2017. Review Manager 5.3 was used for the Meta analysis.

RESULTSSeven RCTs were included. The Meta analysis showed that after 72 hours and 7 days of treatment, the intravenous immunoglobulin (IVIG) group had a significantly higher percentage of children who achieved platelet count >20×10/L than the anti-D group (P<0.05). There were no significant differences in platelet count after 24 hours, 72 hours, and 7 days of treatment between the anti-D (50 μg/kg) group and the IVIG group (P>0.05), and there were also no significant differences in platelet count after 24 hours and 7 days of treatment between the 50 μg/kg and 75 μg/kg anti-D groups (P>0.05). The anti-D group had a significantly greater reduction in the hemoglobin level than the IVIG group after treatment, but did not need transfusion. No children in the anti-D group or the IVIG group experienced serious adverse reactions.

CONCLUSIONSIntravenous injection of anti-D may have a similar effect as IVIG in improving platelet count in children with acute ITP, but it may be slightly inferior to IVIG in the rate of platelet increase after treatment. The anti-D dose of 50 μg/kg may have a similar effect as 75 μg/kg. The recommended dose of anti-D for treatment of ITP is safe.

Humans ; Immunoglobulins, Intravenous ; adverse effects ; therapeutic use ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; blood ; drug therapy ; Rho(D) Immune Globulin ; adverse effects ; therapeutic use