ObjectiveTo investigate the association of HLA-Cw and -DRB1 alleles with psoriasis vulgaris,and to provide a clue to the study into the etiology of psoriasis.MethodsVenous blood samples were obtained from 81 patients with psoriasis vulgaris collected during 2006-2011 at the Department of Dermatology,First Affiliated Hospital of Inner Mongolia Medical College,as well as 100 age- and gender-matched healthy controls.Both the patients and controls are unrelated Mongolia in Inner Mongolia.PCR with sequence-specific primers(PCR-SSP) technique was used to genotype the HLA-Cw and DRB1 loci.ResultsThe patients with psoriasis vulgaris showed a significantly higher frequency of HLA-Cw*06(0.438 vs.0.175,Pc ＜ 0.01) and DRB1*07(0.241 vs.0.110,Pc ＜ 0.012),but a lower frequency of HLA-Cw*04(0.031 vs.0.150,Pc ＜ 0.01 ) and DRB1*04 (0.093 vs.0.235,Pc ＜ 0.01 ) than the healthy controls did.Increased frequencies of HLA-Cw*06 and DRB1*07 alleles were observed in patients with an onset before 40 years of age and those without a family history,together with a decreased frequency of HLA-Cw*04 and DRB1*04 alleles,compared with the healthy controls(Pc ＜ 0.05).The frequency of HLA-Cw*06 allele was significantly higher in patients with a positive family history and patients with an onset of no younger than 40 years of age than in the healthy controls (both Pc ＜ 0.05).ConclusionsHLA-Cw*06 and -DRB1*07 alleles may be susceptibility determinants to psoriasis vulgaris,while HLA-Cw*04 and -DRB1*04 alleles may be protective factors against psoriasis vulgaris,in Mongolia from Inner Mongolia.HLA-DRB1*07 allele may be a susceptibility gene for psoriasis,while HLA-Cw*04 and -DRB1*04 alleles may be protective factors against psoriasis,in patients with an onset before 40 years of age.

Objective:To explore the diagnostic value of serum cytokines and autoantibodies in children with central diabetes insipidus.Methods:A retrospective study was used.A total of 52 children diagnosed with central diabetes insipidus who were hospitalized in Qingdao municipal hospital affiliated to Qingdao Universityfrom January 2016 to January 2020 were selected as the study group, and 60 healthy children with similar gender and age were selected as the control group.The general data of the study group and the control group, 24 h urine volume, urine specific gravity and urine osmotic pressure before the start of water deprivation test, the results of pituitary magnetic resonance imaging (MRI), and the concentration of serum cytokines and autoimmune antibodies were compared to evaluate the value of serum cytokines and autoimmune antibodies in the prediction of central diabetes insipidus.Results:The 24 h urine volume in the study group (201.6 ± 9.9 mL/kg)was significantly higher than that in the control group(9.06±12.5 mL/kg). The urine specific gravity(1.006±0.002) and urine osmotic pressure((116.4±23.5) mOsm/L)were significantly lower than those in the control group((1.013±0.004) and (649.6±72.4) mOsm/L)( t=15.378, 13.298, 45.625, all P<0.001). MRI examination showed that the proportion of disappearance of high signal in posterior pituitary (46.1%(24/52)), the proportion of thickening of the pituitary stalk(32.7%(17/52))and abnormal signal in pituitary(7.7%(4/52)) in study group were significantly higher than the control group(0%(0/0)) (all P<0.05); the levels of serum IL-6, TNF-α, IFN-γ, IL-1β ((117.0±7.8), (234±21.4), (178.6±15.1), (87±6.5) ng/L)and autoimmune antibodies ((654.1±194.3) ng/L) in study group were significantly increased compared to the control((12.5±2.3), (32.5±9.7), (37.8±7.3), (22.6±4.7), (98.0±23.7) ng/L) ( t value were 17.031, 27.490, 15.670, 12.360 and 109.984, respectively; all P<0.05). Conclusion:The concentration of serum cytokines and autoimmune antibodies has a significant clinical value in the evaluation of central diabetes insipidus in children.Combined with head MRI examination, it has a certain value in the diagnosis, treatment and prognosis of central diabetes insipidus in children.

The aim of the present study is to investigate the effect of pulsed electromagnetic field (PEMF) on the activation of human monocytes (THP-1). Cultured THP-1 cells were exposed to PEMF stimulation with radiation of 32Hz or 64Hz respectively, using sinusoidal wave, and 1mT, twice a day, 30 minutes each time, with an interval of 8 hours, for 3 days. Those with 0Hz stimulation served as the controls. Monocytes activation was monitored by measuring both the release of monocyte chemoattractant protein-1 (MCP-1) from monocytes and their adhesion to monolayers of human umbilical vein endothelial cells (HUVECs). The adhesion of THP-1 cells to HUVECs was evaluated by cell counting method. The secretion of MCP-1 from THP-1 cells was detected by ELISA and MCP-1 mRNA expression was assessed by real time quantitative RT-PCR. The data showed that exposure to PEMF with above parameters could significantly inhibit the adhesion of THP-1 cells to HUVECs and decrease the MCP-1 mRNA and protein expression. The results demonstrated that exposure to PEMF of 1mT, 32Hz or 64Hz for 3 days could significantly inhibit the activation of THP-1 cells.
Cell Adhesion ; Cell Line ; Cells, Cultured ; Chemokine CCL2 ; genetics ; metabolism ; Electromagnetic Fields ; Human Umbilical Vein Endothelial Cells ; Humans ; Monocytes ; cytology ; RNA, Messenger ; genetics ; metabolism

Amniotic epithelial cells (AECs) have been expected to be a good cell source for stem cell-based cardiac repair. Activin A signaling is required for cardiac differentiation in human embryonic stem cells (ESCs), and bone morphogenetic protein-4 (BMP-4) is an important regulator that controls stem cell fates. Previous study has established an efficient protocol to generate cardiomyocytes from human ESCs via induction with Activin A and BMP-4. The aim of present study was to test the hypothesis that Activin A and BMP-4 could also induce AECs to differentiate into cardiomyocytes in vitro. Human AECs (hAECs) were isolated from human term placenta by trypsin digestion according to the previous reports. Freshly isolated hAECs were examined to detect the expression of cytokeratin 19 by immunocytochemistry. High-density undifferentiated hAECs at passage 1 were sequential treated with 100 ng/ ml human recombinant Actvin A and 10 ng/ml BMP-4. The expression of cardiac-specific genes was examined before and after in vitro induction of cellular differentiation. Freshly isolated hAEC could express cytokeratin 19, the specific marker of epithelial cells. The data showed that hAECs treated with Activin A and BMP-4 were able to express cardiac-specific genes, including Nkx2.5 and alpha-actinin. Our results demonstrated that Activin A and BMP-4 could induce cardiomyocyte differentiation of hAECs, which might be a novel approach to induce differentiation of AECs into cardiomyocytes-like cells.
Activins ; pharmacology ; Amnion ; cytology ; Bone Morphogenetic Protein 4 ; pharmacology ; Cell Culture Techniques ; Cell Differentiation ; drug effects ; Cells, Cultured ; Epithelial Cells ; cytology ; Humans ; Myocytes, Cardiac ; cytology ; Recombinant Proteins ; pharmacology

OBJECTIVETo research the isolation method, identification and screen for bioactivities endophytic fungi from ginkgo.

METHODEndophytic fungi from ginkgo were separated. By means of microdilution method, activities of endophytes against pathogenic fungi were tested. Then, using DPPH, the antioxidant activities were measured.

RESULTNine strains (16.1%) showed antifungal activities against Candida albicans, Cryptococcus neoformans, Trichophyton rubrum and Aspergillus fumigatus. Among these bioactive strains, the growth of T. rubrum was strongly inhibited by T-1-2-1, as the MIC80 was equal to fluconazole, the positive control. Five strains (8.9%) showed antioxidant activities. Among them sample T-3-2-2 and T-6-5-7 showed the strongest antioxidant activities.

CONCLUSIONEndophytic fungi of ginkgo would be potential and rich resources for drug development.

Antifungal Agents ; pharmacology ; Aspergillus fumigatus ; drug effects ; Candida albicans ; drug effects ; Cryptococcus neoformans ; drug effects ; Fluconazole ; pharmacology ; Fungi ; chemistry ; drug effects ; isolation & purification ; Ginkgo biloba ; microbiology ; Microbial Sensitivity Tests ; Trichophyton ; drug effects

Objective:To screen the neutralizing epitope of enterovirus 71 (EV71) and determine the specific minimum amino acid sequence that triggers immunity for providing a theoretical basis for the development of synthetic peptide vaccines.Methods:EV71 neutralizing antibody-specific binding clones were panned and sequenced using a phage display random 12-peptide library to obtain the key sequences of neutralizing epitopes. A series of peptides containing the key sequences with N-terminal acetylation (AC) and C-terminal linking to Keyhole limpet hemocyanin (KLH) were synthesized. Serum samples were collected after immunizing mice with the modified peptides. Then the immunogenicity of the peptides and the neutralizing activity of serum samples were analyzed by Western blot, ELISA and neutralization test.Results:After three rounds of panning, cloning and sequencing, KQEKDL was identified as the key motif. The serum samples collected from the mice immunized with the modified series of peptides containing key motifs had different degrees of binding ability to EV71 and VP1 protein. The serum samples of mice immunized the synthetic peptide containing only the minimum key motif (AC-KQEKDL-KLH) had the strongest response to the other three peptides and EV71 and the highest neutralizing titer.Conclusions:The EV71 neutralizing epitope was successfully screened using the phage display random peptide library. The key motif of KQEKDL might be the specific minimum amino acid sequence that triggered the immune system. This study provides a theoretical basis for better understanding the immune response mechanism, evaluating the immunogenicity of the antigens and further research and development of polypeptide vaccines.

Complete excision of the left gastroepiploic mesentery and complete dissection the No.4sb lymph nodes is the entrance and one of important steps of splenic hilar lymph node dissection. 4K laparoscopy has a higher level of surgical vision and more subtle degree of local anatomical recognition.Following the concept of mesentery anatomy and with the help of the unique advantages of 4K laparoscopy, the lymph node dissection of left gastroepiploic region is more thoroughly, the operation time is shorter, and there is no bleeding or micro-bleeding. At the same time, the maximum protection of the surrounding important organs and blood vessels, can further improve the radical and safety of the operation. The authors investigate the lymph node dissection along the left gastroepiploic vessels region from intermediate approach in 4K laparoscopic radical gastrectomy.

Objective To investigate the effect of preoperative oral midazolam on postoperative de-lirium in elderly patients with preoperative moderate-severe anxiety undergoing radical resection of colorectal cancer.Methods Eighty elderly patients undergoing laparoscopic surgery for radical resection of colorectal cancer,32 males and 48 females,aged 65-79 years,BMI 21-27 kg/m2,ASA physical status Ⅱ or Ⅲ,the state-trait anxiety inventory(STAI-S)≥38 scores at admission were selected.Patients were divided into two groups using random number method:control group and midazolam group,40 patients in each group.The midazolam group were administrated midazolam 7.5 mg per night till one day before surgery,while the placebo was administrated in the control group.The incidence of delirium 3 days after surgery and the STAI-S scores of one day before surgery were evaluated.The HR and MAP at entry,30 minutes after an-esthesia induction,1 hour,2 hours after anesthesia induction,and 30 minutes after extubation were recor-ded.The total dose of propofol,remifentanil and dexmedetomidine and the using rate of metaraminol were recorded.The visual analog scale scores 30 minutes after extubation,24 and 72 hours after surgery,the u-sing rate of tramadol,and the extubation time were recorded.Results Compared with the control group,the STAI-S scores of one day before surgery,and the incidence of postoperative delirium,the rate of using metaraminol,the VAS scores 30 minutes after extubation and 24 hours after surgery,the rate using of tram-adol were significantly decreased in the midazolam group(P＜0.05).There were no significant differences in total dose of propofol,remifentanil,and dexmedetomidine,extubation time between the two groups.Conclusion Preoperative oral midazolam can effectively reduce the incidence of postoperative delirium in elderly patients with preoperative moderate-severe anxiety undergoing radical resection of colorectal cancer.

Toxoplasma gondii cathepsin C proteases (TgCPC1, 2, and 3) are important for the growth and survival of T. gondii. In the present study, B-cell and T-cell epitopes of TgCPC1 were predicted using DNAstar and the Immune Epitope Database. A TgCPC1 DNA vaccine was constructed, and its ability to induce protective immune responses against toxoplasmosis in BALB/c mice was evaluated in the presence or absence of the adjuvant α-GalCer. As results, TgCPC1 DNA vaccine with or without adjuvant α-GalCer showed higher levels of IgG and IgG2a in the serum, as well as IL-2 and IFN-γ in the spleen compared to controls (PBS, pEGFP-C1, and α-Galcer). Upon challenge infection with tachyzoites of T. gondii (RH), pCPC1/α-Galcer immunized mice showed the longest survival among all the groups. Mice vaccinated with DNA vaccine without adjuvant (pCPC1) showed better protective immunity compared to other controls (PBS, pEGFP-C1, and α-Galcer). These results indicate that a DNA vaccine encoding TgCPC1 is a potential vaccine candidate against toxoplasmosis.
Animals ; B-Lymphocytes ; Cathepsin C* ; Cathepsins* ; DNA* ; Epitopes, T-Lymphocyte ; Immunoglobulin G ; Interleukin-2 ; Mice* ; Peptide Hydrolases ; Spleen ; Toxoplasma* ; Toxoplasmosis* ; Vaccines, DNA*

Genome shuffling methods were explored for Bacillus subtilis strain molecular breeding. Recycling protoplast fusion, recycling transformation and recycling universal transduction were used for genome shuffling in B. subtilis. Four strains with different nutrition-deficiency markers were used as initial strains. After five rounds protoplast fusion, transformation or transduction, the descendant with 4 markers had not been detected, and the rate of descendant with 3 markers were 4.53 x 10(-4), 1.64 x 10(-4), 4.47 x 10(-3), respectively. A computer program was made to simulate the recycling fusion process. Based on simulation result and comparing the genome shuffling result of B. subtilis in this experiment and that of Streptomyces coelicolor reported in references, effective genome shuffling needs a high recombination rate of at least between 10(-3) and 10(-2).
Bacillus subtilis ; classification ; genetics ; DNA Shuffling ; Genetic Techniques ; Genome, Bacterial ; genetics ; Protein Engineering ; Transformation, Bacterial