Objective To evalute the feasibility of a double hand allogroft to reconstuct the limbs'function. Methods A brain dead donor was chosen under the test of blood group in ABO and Rh,HLA match,PRA and the mixed leukocyte culture.The level of the transplantated hands was 5 cm proximal to the wrists The procedure of the allotransplantation was similar to that of the autologous replantation.The immunosuppressants were taken two days before the operation,in the operation and after the operation. The circulation,the vital signs and the rejection were under the supervision. Results The progress was satisfactory. No rejection was found postoperation.After 10 weeks the callus in both ends of radius and ulna was obvious.After 5 months sensation recovered.After 7 months the internal fixation(plates and screws) was removed and tenolysis was done. Now the recipient could use engrafted hands to wash face,make the bed,wear clothes,brush teeth,peel oranges,eat with a spoon,tie the shoelace slowly,pick up a telephoneor and use the conroller of Television. Conclusions With the immunosuppressive agents,the double hands allograft can survive.The growth rate of bone and nerve is faster than that of autologous replantation.

Objective To construct DNA vaccine containing MOMP gene of Chlamydia trachomatis and to observe immune response in mice. Methods Mice of 4 - 6 weeks old were immunized with pcDNA3.1-MOMP or pcDNA3.1 intramuscularly at a dose of 100 μg. Booster immunizations were employed at 2-week interval for two times. Specific antibody in the sera of mice and the level of IFN-γ in murine spleen lymphocyte supernatant were detected by ELISA. The proliferation response of spleen cells was detected by MTT assay. Results Significant specific antibody titers were observed and the highest titer was 1: 1 024 in mice after three times immunization with pcDNA3.1-MOMP. The proli-feration response of spleen cells were significantly higher than that of mice injected with pc DNA3.1. IFN-γ reached(532.0 + 45.4)pg/mL in immunized mice. Conclusion Strong responses of humoral and cellular immunity can be evoked by DNA vaccine of pcDNA3.1-MOMP in mice.

Objective To construct the recombinant plasmid containing the major outer membrane protein(MOMP) gene of Chlamydia trachomatis and expres s MOMP protein in E.coli BL21. Methods The MOMP gene was amplified by polymera se chain reaction from the genome of Chlamydia trachomatis serovar D. The amplif ied fragment was directly inserted into pUCm-T vector and verified by DNA sequen cing. MOMP gene was then subcloned into the prokaryotic expression vector pET-22 b(+). The recombinant protein of MOMP was purified by Ni-NTA affinity chromatogr aphy and identified by SDS-PAGE and Western blot. Results The MOMP gene, which is about 1 200 bp, was successfully amplified and cloned. The DNA sequence of t he cloned MOMP gene was the same as that published by the GenBank. SDS-PAGE anal ysis showed that the relative molecular weight of this fusion protein was about 47 kDa which was consistent with the theoretically predicted value, and the spec ificity of this recombinant protein was confirmed by Western blot. Conclusions The MOMP gene of Chlamydia trachomatis was successfully cloned and expressed in the prokaryotic expression system, which may lay the foundation for the developm ent of Chlamydia trachomatis vaccine.

Objective To evaluate the clinical effect of umbilical cord mesenchymal stem cells (MSCs) for treatment of chronic dilated cardiomyopathy complicated with systolic heart failure.Methods Fifty-nine patients with chronic dilated cardiomyopathy and systolic heart failure in Henan Provincial People's Hospital from December 2013 and December 2014 were selected and divided into treatment group (n =30)and control group (n =29).All patients were given routine drug treatment.Based on this,the patients in treatment group were given umbilical cord MSCs 20mL by intracoronary transplantation;the patients in control group were given the same volume of saline.The changes of cardiac function,left ventricular ejection fraction (LVEF),left ventricular end diastolic diameter(LVEDD),6 minutes walking distance,the amino-terminal pro-brain natriuretic peptide (NT-proBNP) levels,the re-hospitalization rate,mortality rate of patients in the two groups were observed and compared at 1 month and 6 months after treatment.Results The cardiac function grading of the New York Heart Association (NYHA)of patients in the two groups were improved at 1 month and 6 months after treatment (P ＜ 0.05);the NYHA cardiac function grading of patients in the treatment group was significantly better than that in the control group at 1 month and 6 months after treatment (x2 =12.64,16.75;P ＜ 0.05).There was no statistic difference in LVEDD,LVEF,serum level of NT-proBNP and 6-min walking distance of patients between the two groups before treatment (P ＞ 0.05).There was statistic difference in LVEDD,LVEF,serum level of NT-proBNP and 6-min walking distance of patients in the two groups before treatment and 1 month,6 months after treatment(P ＞ 0.05).There was no statistic difference in LVEDD of patients between the two groups at 1 month after treatment (P ＞ 0.05);the LVEF,6-min walking distance of patients in the treatment group were significantly higher than those in the control group,but the serum level of NT-proBNP of patients in the treatment group was significantly lower than that in the control group at 1 month after treatment (P ＜ 0.05).At 6 months after treatment,the LVEDD,serum level of NT-proBNP of patients in the treatment group were significantly lower than those in the control group,but the LVEF and 6-min walking distance of patients in the treatment group were significantly higher than those in the control group (P ＜ 0.05).The mortality rate of patients in the treatment group (6.67％,2/30) was significantly lower than that in the control group (24.14％,7/29) (x2 =4.99,P ＜ 0.05).The re-hospitalization rate of patients in the treatment group and control group was 16.67％ (5/30) and 31.031.(9/29) respectively;there was no statistic difference in the re-hospitalization rate of patients between the two groups (x2 =1.68,P ＞ 0.05).Conclusion Umbilical cord MSCs can improve the cardiac function and cardiac remodeling,reduce the mortality rate of patients with chronic dilated cardiomyopathy complicated with systolic heart failure.

BACKGROUND: Both therapeutic effects of stem cell transplantation and differentiation mechanism remain controversial;however, plentiful clinical studies have indicated that stem call transplantation might improve heart function.OBJECTIVE: To evaluate the efficacy of intracoronary autologous bone marrow mononuclear call transplantation for the treatment of chronic heart failure.DESIGN: Case analysis. PARTICIPANTS: A total of 80 patients with chronic heart failure who were selected from Henan People's Hospital from 2003 to 2008 were divided into two groups: autologous bone marrow mononuclear call transplantation group (n=48) and conventional drug therapy group (n=32). There were no significant differences in sex, age, accepting conventional drug therapy,echocardiogram before transplantation, plasma brain natriuretic peptide level, and myocardial perfusion imaging between the two goups (P>0.05).METHODS: A one-year following up before and after cell transplantation, conventional drug therapy was performed in the two groups. Based on conventional drug therapy, autologous bone marrow mononuclear call suspension (10 mL) was slowly poured into coronary artery, and the cell number was adjusted to (3.1±1.6)×10 8. MAIN OUTCOME MEASURES: Changes of heart function were measured using ultrasound apparatus; plasma brain natriuretic peptide levels were detected using double antibodies immumofluorescence method; area of myocardial perfusion defect region was detected using single photon emission computed tomography.RESULTS: Patients in the two groups finished the one-year follow-up, and adverse effects and complications were not found before and after cell transplantation. ①Compared to before cell transplantation, end-systolic volume (ESV) of the left ventricle was significantly decreased in the autologous bone marrow mononuclear cell transplantation group after one year (P < 0.05), but ejection fraction (EF) of the left ventricle was significantly increased (P < 0.05). However, both ESV and EF were not changed in the conventional drug therapy group (P> 0.05). Compared to conventional drug therapy group, ESV and EF were changed significantly in the autologous bone marrow mononuclear call transplantation group after one year (P < 0.05). ②Compared to before cell transplantation, plasma brain natriuretic peptide levels were significantly decreased in the two groups (P < 0.05 or 0.01). Area of myocardial perfusion defect region was significantly decreased (P < 0.05 or 0.01), and the area change in the autologous bone marrow mononuclear call transplantation group was significantly greater than conventional drug therapy group (P < 0.05).CONCLUSION: Autologous bone marrow mononucleer call transplantation is safe and feasible for the treatment of chronic heart failure, and it also can remarkably improve heart function and myocardial perfusion within one year.

Objective:To observe the changes of metabolic intensity value of points on the face before and after needling bilateral Hegu (LI 4) in healthy people and provide scientific basis for association between Hegu (LI 4) and face/mouth. Methods:A total of 45 healthy college students were selected in this study. Using medical thermography and Pennes bio-heat transfer model, the infrared thermograph images on the face before and after needling bilateral Hegu (LI 4) were collected to observe the distribution of metabolic intensity value on the face before acupuncture and changes in these values after needling bilateral Hegu (LI 4). Results:Before acupuncture, Cuanzhu (BL 2) had the maximal metabolic intensity value. Its mean value was (0.71±0.23) W. Quanliao (SI 18) had the minimal metabolic intensity value. There were no left-right statistical significances in metabolic intensity values. After needling bilateral Hegu (LI 4), the metabolic intensity values of most points on the face were increased. Kouheliao (LI 19) obtained the maximal increase: 0.35 W on average; and Yangbai (GB 14) obtained the minimal increase: 0.08 W on average. Conclusion:Points on both sides in healthy people have good symmetry in metabolic intensity value. After needling bilateral Hegu (LI 4), the metabolic intensity values of points on the face were increased, especially points around the lips, which accords with the pathway of the Large Intestine Meridian on the head and face. This provided some scientific foundation for the association between Hegu (LI 4) and face/mouth.

Objective To investigate the specific humoral immune response and cellular immune response induced by DNA vaccine with Neisseria gonorrhoeae porin B (PorB) fused with B subunit of Escherichia coli heat-labile enterotoxin B (LTB) in mice. Methods Target genes of porB, ltB and ltB-porB were amplified by polymerase chain reaction (PCR) and cloned into eukaryotic vector pcDNA3.1(-). The recombinants were identified by PCR, enzyme digestion and DNA sequencing.The vectors were transfected into Hela cells, and expressed proteins were checked by cytoimmunofluorescence. Female BALB/c mice were intranasally immunized with recombination vectors. The humoral immune response and cellular immune response were detected by enzyme linked immunosorbent assay (ELISA) and methyl thiazolyl tetrazolium (MTT) colorimetric assay. The expressions of recombination vectors in intranasal mucosal tissues of the immunized mice were detected by immunohistochemistry. The means between groups were compared by analysis of variance. Results All the three recombinants were expressed in Hela cells and intranasal mucosal tissues. The PorB specific IgG in serum and sIgA in vaginal secretions in DNA vaccine immunized mice were significantly higher than those in controls (P＜0.01 ; P＜0.05). Moreover, the sIgA level in pcDNA3.1 (-)/ltB-porB group was higher than that in peDNA3, 1(-)/porB group (P＝0. 002). The levels of interferon-gamma (IFN-γ) and interleukin-4 (IL-4) in the supernatants and stimulation index (SI) of spleen lymphocyte culture in pcDNA3, 1(-)/porB group were (170.04±23.89) pg/mL, (114.68±14.27) pg/mL and 1. 68±0.19, respectively; and those in pcDNA3, 1(-)/ltB-porB group were (161.42±27.50) pg/mL, (124.16±19.04) pg/mL and 1.73±0.28, respectively; which were both higher than those in pcDNA3.1(-)/ phosphate buffered saliae (PBS) group (P＜0. 01; P＜0.05) and pcDNA3.1 (-)/ltB group (all P＜0.05), while there was no significant difference between pcDNA3.1 (-)/ltB-porB group and pcDNA3. 1 (-)/porB group (0. 998, 0. 696, 0. 994; all P＞0.05). Conclusions The constructed DNA vaccines are all successfully expressed in Hela cells and murine intranasal mucosal tissues. The mucosal immunization of the vaccines [pcDNA3. 1 (- )/porB and pcDNA3.1 ( -)/ltBporB] could induce humoral immune response and cellular immune response, especially mucosal immune response. It is confirmed that mucosal adjuvant LTB could promote PorB to induce higher level of mucosal immune response in mice.

Objective:To analyze the changes in biological characteristics including infectivity, growth and pathogenicity of Chlamydia muridarum ( Cm) after serial passage in vitro in special conditions in order to provide reference for screening attenuated live vaccines and virulence-related genes. Methods:Wild-type Cm strain (G0) was cultured for several passages using conventional cell culture method under alternate unassisted and assisted culture conditions. Then, the 28th generation (G28) of Cm was selected and compared with the parental G0 strain in terms of centrifugation dependence, attaching ability, intracellular growth curve, plaque size and fallopian tube lesions after genital tract infection in a mouse model. Results:Compared with the parental G0 strain, the G28 strain showed significantly decreased dependence on centrifugation during cell infection ( P<0.05) and increased attachment capacity to cells ( P<0.05). No significant differences were observed in the growth curves 32 h after cell infection or in the plaque sizes between the parental G0 and G28 strains. In the in vivo virulence test, fallopian tube lesions were observed in 87.5% of G0-infected mice and 37.5% of G28-infected mice ( P<0.05). Conclusions:Compared with the parental G0 strain, the G28 strain showed significantly enhanced in vitro infection ability, but decreased in vivo pathogenicity, which brought hope for further identification of virulence genes, isolation of attenuated strains with single genotype and development of live attenuated Chlamydia vaccines.

Objective To investigate the compliance of secondary prevention and the relationship with the long-term outcomes in patients undergoing percutaneous coronary intervention(PCI).Methods 589 patients undergoing PCI were followed-up,and factors including major adverse cardiac events(MACE)),smoking status and the usage of antiplatelet agents,angiotensin converting enzyme inhibitor(ACEI)/angiotensin Ⅱ receptor blocker(ARB),statins,beta blocker,calcium channel blocker and nitrates were recorded.Results The average follow-up time was 18.92 months.At discharge,588 patients(99.83%)were prescribed clopidogrel for(7.89±4.96)months;there were 31 patients(5.26%)who completely discontinued antiplatelet therapy during follow-up.At discharge,the prescription rate of aspirin,ACEI/ARB,beta blocker,statins,calcium channel blocker and nitrates was 98.98%,41.94%,63.50%,83.02%,19.69%and 46.52%respectively,whereas at follow-up,these were decreased to 94.4%,35.99%,55.86%,65.89%,17.49%and 35.31%.At follow-up,there were still 105 current smokers(17.83%).Complete cessation of antiplatelet therapy and current smoking were related to the increased risk of non-fatal myocardial infarct(9.68%v.s.1.08%,P＜0.01);smoking(4.76%v.s.0.83%,P＜0.01)andMACE(19.35%v.s.6.45%,P＜0.01);smoking(11.43%v.s.6.20%,P＜0.05).Conclusion Most patients can adhere to secondary prevention during follow-up,however,the compliance with secondary prevention should be improved further.Cessation of antiplatelet therapy and current smoking contribute to poor prognosis.

Isopentenyl flavonoids are a class of characteristic components in Sophora flavescens Ait. (S. flavescens). They have biological activities such as anti-tumor, anti-bacteria, anti-inflammol/ Lation and anti-oxidation. In this paper, the structural types, toxicology and pharmacological effects of isopentenyl flavonoids from S. flavescens were briefly reviewed. Furthermore, the worth of further study on pharmacokinetics, pharmacodynamics, toxicology, action targets, molecular mechanisms and structure-function relationships of isopentenyl flavonoids were proposed. The deep exploration on functional characterastics of isopentenyl flavonoids of S. flavescens and their application on development of innovative drugs are of great significance to further improve the added value of isopentenyl flavonoids and expand their application fields.