Risk factors of cardiovascular and cerebrovascular diseases in young patients with hyperhomocysteinemia / 中国基层医药

Objective:To investigate the risk factors of cardiovascular and cerebrovascular diseases in young patients with hyperhomocysteinemia.Methods:A total of 260 patients younger than 45 years old who received treatment at the Department of Emergency, Seventh Medical Center, Chinese PLA General Hospital from January 2018 to January 2019 were included in this study. Among these patients, 126 patients with serum homocysteine levels ≥ 15.0 μmol/L were included in the hyperhomocysteinemia group, and 134 patients with serum homocysteine levels < 15.0 μmol/L were included in the control group. Height, body weight, body mass index, blood pressure, homocysteine, fasting blood glucose, blood uric acid, total cholesterol, triacylglycerol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and N-terminal B-type natriuretic peptide were determined in each group. Changes in risk factors of cardiovascular and cerebrovascular diseases were compared between the hyperhomocysteinemia and control groups.Results:There were significant differences in body mass index [(26.42 ± 3.54) kg/m 2vs. (22.14 ± 3.22) kg/m 2, t = 10.21, P = 0.016], blood uric acid [(308.71 ± 78.44) μmol/L vs. (285.05 ± 92.09) μmol/L, t = 2.22, P = 0.027], the incidence of coronary heart disease (73/126 vs. 61/134, χ2 = 4.00, P = 0.045) and the incidence of stroke (19/126 vs. 6/134, χ2 = 8.39, P = 0.004) between the hyperhomocysteinemia and control groups. There were no significant differences in fasting blood glucose, blood lipid level, N-terminal B-type natriuretic peptide, and the incidences of diabetes mellitus and hypertension between the two groups (all P > 0.05). Conclusion:The related risk factors of cardiovascular and cerebrovascular diseases increase in young patients with hyperhomocysteinemia. The incidences of coronary heart disease and stroke are very high, and therefore timely intervention should be carried out.

Research progress of mesenchymal stem cells therapy for right heart failure associated with pulmonary arterial hypertension / 中华胸心血管外科杂志

Pulmonary hypertension is a progressive disease characterized by pulmonary vascular remodeling and eventually develops into right heart failure, which seriously affects the quality of life and safety of patients. Traditional drug therapy can alleviate disease progression, but the prognosis is poor.Mesenchymal stem cells have been shown to be effective in experimental pulmonary hypertension and right heart failure, which is an important research direction in the future.In this paper, the research progress of mesenchymal stem cells in pulmonary hypertension and right heart failure is reviewed.

Benefits and related problems of influenza immunization for health care workers / 中华实用儿科临床杂志

Vaccination is the most effective way to prevent influenza and severe outcoming caused by influenza viruses.Health care workers(HCW) are exposed to patients with influenza and they are at high risk of occupationally acquired influenza and of causing nosocomial infection among patients,increasing the incidence rate,the risk of severe and death of patients.Improving the influenza immunization in HCW can not only reduce the prevalence of themselves and keep a weel-oiled of health care facilities during the influenza seasons,but also reduce the risk of severe and death among patients and increase the influenza vaccine uptake in whole population.At present,the influenza immunization coverage of HCW is low.The obstacles and myths of influenza vaccine are barriers for vaccine uptake among HCW.The various strategies are critical in order to improve the influenza coverage rates of HCW.

In vitro expression of human cytomegalovirus UL148 RNA and prediction of its functional sites / 中华微生物学和免疫学杂志

Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.

Effective inhibition of human cytomegalovirus UL148 gene expression by external guide sequences in vitro / 中华实验和临床病毒学杂志

Objective@#To investigate the UL148 gene function of human cytomegalovirus (HCMV) low passage clinic isolate and new strategies for anti-HCMV treatment, the DNA-based external guide sequences (EGSs) were designed to inhibit UL148 RNA expression.@*Methods@#UL148 RNA secondary structure was analyzed by RNA structure technique, an appropriate region was chosen for DNA-based EGS57 synthesis, targeted the UL148 RNA. The M1RNA and UL148 RNA were generated by PCR for transcription in vitro. The UL148 RNA and M1RNA were transcribed in vitro under the function of T7 RNA polymerase. The UL148 was labelled by 32P. The cleavage reactions were carried out by mixing up EGS, M1RNA with UL148 RNA for 1 h. The products were separated by urea denaturing polyacrylamide gel electrophoresis and detected with Typhoon Phosphor Imager.@*Results@#UL148 RNA ranged from 58 to 72 sites was the binding position, and 57 was a cleavage site. EGS57 was designed and synthesized. EGS57 was combined with UL148 RNA to form the natural substrate of M1RNA. UL148 RNA and M1RNA were synthesized through T7 RNA polymerase catalyzing, and the products were conformed. After cleaving reactions, DNA-based EGS57 was shown to be able to cleave UL148 RNA efficiently in vitro by a complex with M1RNA.@*Conclusions@#UL148 RNA was cleaved efficiently by EGS57, and the cleaving site was conformed as expectation. It will provide the gene silent tool effectively for further study the function of UL148 gene.

Construction of a murine model of chronic graft-versus-host disease by haplo-identical spleen cell infusion / 中华器官移植杂志

Objective For providing experimental platform of chronic graft-versus-host disease (cGVHD),to establish a mouse model by haplo-identical spleen cell infusion.Methods The donor male mice (Balb/cH-2d) and the recipient (Balb/c C57BL/6) F1 H2-d/b (CB6F1) female mice were randomly divided into four groups:3 experimental groups injected with 3 107,6 107 and 9 107 spleen cells,respectively,while the control group received RPMI 1640 solution.H-2d and H-2b were checked to analyze the chimerism in bone marrow cells.Body mass,figure,cutaneous manifestation and survival of recipient mice were observed and scored every 3 days.Pathologic changes of target organs were observed and scored.Results Injection of 6 107 and 6 107 splenocytes in the recipient mice resulted in a chronic disease with a low level of parental cell engraftment steadily.As compared with 3 107 group,the incidence of cGVHD in 6 107 and 9 107 groups were significantly increased (P ＜0.01).But there was no significant difference between 6 107 and 9 107 groups (P＞0.05).Conclusion A murine model of cGVHD after haplo-identical spleen cell infusion of donor is successfully established by injection of 6 107 and 9 107 spleen cells.

Analysis of nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cy-tomegalovirus strains in Guangzhou / 中华微生物学和免疫学杂志

Objective To analyze the nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cytomegalovirus ( HCMV) strains isolated from infants in Guangzhou province. Methods The low passage strains of HCMV were isolated from urine samples of 10 infants with HCMV in-fection in Guangzhou province and identified by multiplex PCR.The UL138 genes were amplified, cloned and identified with sequencing.The sequences were analyzed together with the homologous sequences of 10 clinical isolates published in GenBank.The sequences of UL138 genes were analyzed by using bioinformatics softwares for investigation of the post-translational modification sites, isoelectric points and second structures of UL138 proteins.Results Three low passage strains of HCMV ( D2, D3 and D52) were isolated from in-fants with congenital HCMV infection.The complete sequences of UL138 genes of the three strains were sub-mitted to GenBank after sequencing identification with the GenBank accession numbers of DQ180375, DQ180387 and DQ180359, respectively.The UL138 gene sequences of the three clinical isolates were high-ly conservative.Among the 841 base pairs of the UL138 gene sequences, mutations were identified in 16 sites with base substitution, no any insertion and deletion mutation was found.The 16 mutations resulted in 7 amino acid changes.No additional or deleted sites were found with regard to the post translational modifi-cation sites of UL138 protein in all clinical isolates except the Toledo strain.The isoelectric point of UL138 protein was 6.51 for all clinical isolates.Conclusion The UL138 genes and the deduced amino acid se-quences of HCMV strains isolated from infants in Guangzhou were highly conservative, regardless of the poly-morphism of UL138 gene.This study paved the way for further investigation on HCMV infection and its path-ogenic mechanism.

Influence of Annao tablet on expression of transforming growth factor beta 1 in acute graft-versus-host disease murine / 实用医学杂志

Objective To study the influence of Annao tablet on the expression of transforming growth factor beta 1 (TGF-β1) in acute graft-versus-host disease (aGVHD) murine and to explore the interventional mechanism of TGF-β1 on aGVHD. Methods Hematopoietic stem cells of male Balb/c mice were transplanted to female C57BL/6 mice for the development of aGVHD murine model. Recipient mice were divided into Annao group and blank group randomly and respectively administrated with Annao soup (a kind of Chinese herb) and 0.9% sodium chloride intragastrically. Clinical symptom, survival time and body weight were recorded at 14th and 30th day and some sections of liver, small bowel and skin were taken for histological changes. Serum level of TGF-β1 were measured by enzyme-labeled immunosorbent assay (ELISA), splenocyte protein of TGF-β1 by Western Blot and TGF-β1 mRNA by fluorescent quantitation polymerase chain reaction (PCR). Results Serum level of TGF-β1 in both groups had no statistical difference (P = 0.305), but it rose to (148.31 ± 7.95) ng/mL at 14th day and (183.48 ± 5.91) ng/mL at 30th day in Annao group, which had significant difference when compared with that in blank group (P = 0.000). IOD/IODβ-actin value of TGF-β1 protein in Annao group was 0.33 ± 0.05 at 14th day and 0.56 ± 0.04 at 30th day, which was higher than that in blank group (P = 0.000) and the expression of TGF-β1 mRNA of splenocyte in Annao group was 1.24 ± 0.04 at 14th day and 2.14 ± 0.33 at 30th day which was much higher than that in blank group (P = 0.000). Conclusion Annao tablet helps to relieve symptoms of acute GVHD by raising serum level of TGF-β1 and intensifying expression of protein of TGF-β1 and its mRNA.

Haploidentical hematopoietic stem-cell transplantation for acute myeloid leukemia in first relapse after complete remission by standard induction chemotherapy / 中华器官移植杂志

Objective To investigate the therapeutic effects of haploidentical hematopoietic stem-cell transplantation (Haplo-PBSCT) for acute myeloid leukemia in first relapse after complete remission by standard induction chemotherapy. Methods Eighty-nine cases of AML in first relapse after complete remission by standard DA/Hi-Ara-C regimens induction chemotherapy were evaluated retrospectively. Fiftythree cases were grafted by haplo-PBSCT and 26 cases were treated with iDA/Mid-Ara-C or MA/ Mid- Ara-C agents. Results The second remission rate in haplo-PBSCT group and continuous chemotherapy group was 86. 7 % (46/53 cases) and 38. 1% (9/23 cases) respectively (P＜0. 01). Survival postprogression (SPP) at 36th month was 43. 4 % (23/53 cases) in haplo-PBSCT group and 11.5 % (3/26 cases) in continuous chemotherapy group (P ＜ 0. 05). Conclusion Haplo-PBSCT could significantly increase the second remission rate and prolong the survival time of patients with acute myeloid leukernia in first relapse after complete remission by standard induction chemotherapy.

Antitumor effect of allogeneic NK cells on human multi-drug resistant nasopharyngeal carcinoma cells subcutaneously transplanted in BALB/c nude mice / 中国肿瘤生物治疗杂志

Objective:To study the inhibitory effect of allogeneic natural killer(NK) cells on subcutaneously transplanted human multi-drug resistant nasopharyngeal carcinoma cells(CNE2/DDP) in BALB/c nude mice.Methods:Human leucocyte antigen(HLA) genotypes of CNE2/DDP cells and the genotypes of inhibitory killer cell immunoglobulin-like receptor(KIR) in NK cells(isolated from 3 healthy persons by immuno-magnetic microbead technique) were analyzed by PCR-SSP.Twelve BALB/c nude mice were evenly divided into 2 groups:the control group and the treatment group.Mice in the treatment group were injected subcutaneously with 1?106 CNE2/DDP cells together with 3?107 NK cells via the tail veins;mice in the control group were injected with 1?106 CNE2/DDP cells subcutaneously.The tumor formation time,tumor formation rate and changes of tumor size were observed.Three weeks after tumor formation,all the mice were killed and human NK cells in peripheral blood were analyzed by flow cytometry;the tumors were weighed and the tumor inhibitory rates were calculated.Results:The HLA genotypes of CNE2/DDPcells were A2,24,B18,35,Cw4,and 7;the KIR genotypes of the 3 healthy persons were KIR2DL1,KIR2DL3,KIR3DL1,and KIR3DL2.There were mismatches between the KIRs expressed in NK cells and HLA class Ⅰ molecules expressed in the CNE2/DDP cells.NK cells obviously inhibited the growth of CNE2/DDP xenograft in nude mice.The tumor formation periods of control group and NK cell group were(17.17?1.17) d and(24.83?1.47) d,respectively(P