Expression of PD1/PD-L1 in gastric carcinoma and its correlation with clinicopathological characteristics and prognosis / 临床与实验病理学杂志

Purpose To investigate the expression of pro-grammed death1 (PD-1), programmed death-ligand1 ( PD-L1) in gastric carcinoma and its correlation with clinicopathological characteristics and prognosis. Method Immunohistochemistry was used to dectect the expression of PD-1/PD-L1 in 75 cases of gastric cancer and adjacent tissues of cancer, without chemother-apy and radiotherapy from 2011 to 2016 in The Affiliated Jiangsu Shengze Hospital of Nanjing Medical University. To analyzed the relationship between expression of PD-1/PD-L1 in gastric carci-noma and clinicopathologic characteristics, and analyzed rela-tionship of overall survival time in all patients. Result The tumor cells expressed 37. 3% (28/75) PD-L1 positive rate in 75 cases, which associated with age, tumor diameter and differ-entiation. PD-1 positive rate in the tumor infiltrating lympho-cytes was 46. 7% (35/75), which associated with pTNM stag-ing. The positively rate of PD-L1 expression in tumor cells and PD-1 expression in the tumor infiltrating lymphocytes were sig-nificantly higher than that in the adjacent tissues of carcinoma (P＜0. 001). Kaplan-Meier and Long-rank results showed the overall survival time was significantly related with tumor diame-ter, differentiation, invasion depth, metastasis and pTNM stag-ing ( P ＜0. 05), but unrelated with PD-1 and PD-L1 expres-sion. But if the follow-up time was 14 months, the prognosis of PD-L1 positive patients were significantly lower than that of the negative ones. Conclusion Expression of PD-1/PD-L1 in cancer cells are significantly higher than that in para-carcinoma tissue. Positive expression of PD-L1 in tumor cells may contrib-ute to evaluation of short-term prognosis, and has nothing to do with the overall survival rate.

Epidemiological survey of childhood asthma in 2010 in urban Baotou, China / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the prevalence rate of childhood asthma in 2010 in urban Baotou, China, as well as the characteristics of attacks and the status of diagnosis and treatment of childhood asthma.</p><p><b>METHODS</b>More than 10 000 children (0-14 years) were selected from 3 secondary schools, 3 primary schools, 6 kindergartens, and 4 community vaccination sites in urban Baotou by cluster random sampling between September 2009 and August 2010. A standardized preliminary questionnaire was used for screening out suspected cases, which were then confirmed or excluded by a clinician; the confirmed cases underwent further questionnaire survey. Double entry and validation was adopted for all data using Epi-Info software, and analysis was performed using SPSS 13.0.</p><p><b>RESULTS</b>A total of 11 323 children were surveyed. Asthma was diagnosed in 127 cases (including 121 children with typical asthma and 6 children with cough variant asthma), with a prevalence rate of 1.12%. The prevalence rate of asthma in male children was significantly higher than that in female children (1.51% vs 0.72%; P<0.01). The prevalence rate of asthma in 2010 was significantly increased compared with that in 1990 (0.55%) and 2000 (0.88%) (P<0.05). Systemic glucocorticoid use decreased significantly from 60.2% in 2000 to 25.9% in 2010 (P<0.01); inhaled corticosteroid use increased significantly from 13.6% in 2000 to 85.8% in 2010 (P<0.01); antibiotic use decreased from 98.1% in 2000 to 66.9% in 2010 (P<0.01). The multivariate logistic regression analysis showed that family history of allergy, allergic rhinitis, chronic cough, and recurrent respiratory tract infection were independent risk factors for childhood asthma.</p><p><b>CONCLUSIONS</b>The prevalence rate of childhood asthma in urban Baotou shows an increasing trend. Inhaled corticosteroids have been widely used.</p>

Changes and clinical significance of CD8(+) T cell subset in patients with aplastic anemia, myelodysplastic syndrome and acute myeloid leukemia / 中国实验血液学杂志

This study was purposed to detect the balance and the activity change of cytotoxic T cell subsets in aplastic anemia (AA) patients, myelodysplastic syndrome (MDS) patients and acute myeloid leukemia (AML) patients, and to explore the cellular immune mechanism for abnormal hematopoiesis of the three diseases, so as to provide experimental basis for the choice of clinical treatment. The proportion of the cytotoxic T cells and part of the T-cells subsets in peripheral blood were detected by flow cytometry in 35 cases of MDS, including 19 refractory anemia (MDS-RA), 16 refractory anemia with excess blasts (MDS-RAEB), 17 AA, 15 AML patients and 10 normal donors respectively. The results showed that compared with the control group, the percentage of Tc1, Tc1/Tc2, CD8(+)HLA-DR(+), CD3(+)CD8(+)CD28(+), CD8(+)CD45RO(+) cells was significantly higher and the percentage of CD8(+)CD45RA(+) was significantly lower in AA and MDS-RA group. There was no difference in the percentage of Tc2 cells between AA/MDS-RA and normal controls; the percentage of CD8(+)CD45RO(+) cells was significantly higher and the percentage of Tc1, CD3(+)CD8(+)CD28(+), CD8(+)HLA-DR(+) was significantly lower in MDS-RAEB group, the percentage of CD8(+)CD45RA(+) was lower but the difference was not significant, and there was no difference in the percentage of Tc, Tc1/Tc2 cells between MDS-RAEB group and the control group. The percentage of Tc2 cells was significantly higher and the percentage of other parameters was significantly lower in AML group than those of normal controls. It is concluded that the cellular immune statuses in AA, the different stages of MDS and AML are different. In AA and the early stage of MDS, the balance of Tc1/Tc2 shifts to Tc1, and the activation of T-cell subsets increases. In the late stage of MDS and AML, the balance of Tc1/Tc2 shifts to Tc2, the activation of T-cell subsets decreases. The former may be closely related to bone marrow failure while the latter may be one of the important mechanisms in which the malignant clones escape from immune effect.

Early apoptotic changes in human spermatozoa and their relationships with conventional semen parameters and sperm DNA fragmentation / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To investigate whether early apoptotic changes in spermatozoa can be significant markers for sperm quality.</p><p><b>METHODS</b>Two early apoptotic changes in the semen of 56 men were assessed using Annexin V (AN)/propidium iodide (PI) staining for phosphatidylserine externalization and JC-1 staining for mitochondrial membrane potential (MMP). The results were compared with conventional semen parameters and DNA fragmentation identified using the TUNEL assay.</p><p><b>RESULTS</b>The different labeling patterns in the bivariate Annexin V/PI analysis identified four distinctive spermatozoa populations. The percentage of AN(-)/PI(-) spermatozoa positively correlated with conventional semen parameters and MMP, but negatively correlated with TUNEL (+) spermatozoa. As for the AN(-)/PI(+) fraction, we found an opposite result in comparison to AN(-)/PI(-) spermatozoa. The level of early apoptotic AN(+)/PI(+) spermatozoa negatively correlated with MMP and sperm motility. The level of late apoptotic AN+/PI+ spermatozoa negatively correlated with conventional semen parameters and MMP, and positively correlated with TUNEL (+) spermatozoa. MMP positively correlated with conventional semen parameters, but negatively correlated with TUNEL (+) spermatozoa.</p><p><b>CONCLUSION</b>Although early apoptotic AN+/PI(-) spermatozoa only negatively correlates with sperm motility, the differences in proportion of each subpopulation of spermatozoa (especially, the percentage of AN(-)/PI(-) spermatozoa), and decreased MMP might be significant markers for diagnosing male infertility. They possibly bring additional information to predict the outcome of in vitro fertilization.</p>

Deletion of spiramycin 3-O-acyltransferase gene from Streptomyces spiramyceticus F21 resulting in the production of spiramycin I as major component / 生物工程学报

Spiramycin (SP) belongs to the 16-member macrolide antibiotics. It contains three components,namely SP I, SP II and SP III, which differ structurally in the acylation moieties on the C3 of the lactone. The SP I component contains a hydroxyl group at C3. SP II, and SP III are formed by further acetylation or propionylation of the C3 of SP I, by the same 3-O-acyltransferase (3-O-AT) . The study focused on simplifying spiramycin components. Theoretically, disruption/deletion of the 3-O-AT gene will reduce/stop the acylation of SP I to SP II and SP III. In this study, degenerated primers were designed according to the conserved regions of 3-O-acyltransferase, MdmB and AcyA in the medicamycin and carbomycin producers of S. mycarofaciens and S. thermotolerans, respectively, and an 878bp DNA fragment was amplified from the spiramycin-producer of S. spiramyceticus F21. Blast analysis of the 878bp DNA fragment suggested that it encoded the 3-O-acyltransferase (3-0-AT, sspA) gene for spiramycin biosynthesis. The flanking regions of this 878bp DNA fragment were then amplified by single-oligonucleotide-nested PCR, and a total of 4.3 kb DNA was obtained (3457nt among the 4.3kb fragment was sequenced, and deposited in GenBank DQ642742),covering the whole putative 3-O-acyltransferase gene, sspA. The sspA was then deleted from the S. spiramyceticus F21 genome by double cross-over homologous recombination, mediated by temperature-sensitive plasmid pKC1139. A comparison was done of the components of spiramycins produced by the sspA-deleted mutant strain with that of the parent strain by HPLC analysis, which showed that sspA-deleted mutant produced SP I (72%), SP II (18%), and SP III (9.6%), whereas parent strain produced SP I (7.8%), SP II (67%), and SP III (25%), respectively, demonstrating the role of ssp A in the acylation of SP I into SP II and SP III. The ssp A-deleted mutant strain obtained in this study may be used for the production of SP I, or may serve as a good starter for the construction of spiramycin derivatives.

Cloning and expression of human interleukin-26 in Escherichia coli / 生物工程学报

To clone human interleukin-26 (hIL-26) and express it in E. coli efficiently. Two pairs of primers were synthesized according to the hIL-26 gene reported on GenBank. The hIL-26 gene was cloned by nest PCR following the first round RT-PCR from human peripherial blood monocytes total RNA, and then the PCR product was cloned into pMD18-T vector. Colony PCR, restriction analysis and sequence analysis showed that the gene cloned was the same as the reported hIL-26. The recombinant was cut with BamHI and EcoR I to obtain the hIL-26 fragment, and then the fragment was inserted into pBV220 which was cut with the same enzymes. The recombinant expression vector was induced to express hIL-26 at 42 degrees C, SDS-PAGE analysis showed that the recombinant protein accounted for up to 20% of the whole protein of E. coli, and the protein was also confirmed by Western blotting. Purity of the protein was found to be above 90% after purified with molecular sieve. After renaturalized with glutathione buffer, the promoting effect of it on the production of IFN-y in PBMC was detected by RT-PCR. A recombinant bacterial strain for expressing hIL-26 with biological activity was constructed successfully.

Presence of autoantibodies in sera of patients with sporadic idiopathic hypoparathyroidism / 中华检验医学杂志

Objective To explore whether autoreactive antibody presents in patients with sporadic idiopathic hypoparathyroidism(sIHP).Methods The subjects including 26 patients with sIHP and 112 genealogical members as well as 60 age-and sex-matched healthy controls.Anti-parathyroid antibodies in the sera were assayed by indirect immunofluorescence.The levels of calcium,phosphorus and magnesium as well as intact parathyroid hormone(iPTH)in the sera were tested.Results Positive autoantibodies against parathyroid tissue were demonstrated in 10 patients(38%)with sIHP,significantly higher than that of in genealogical members(10%,?~2=13.42,P

Selection of Cd~(2+) Binding-peptides and Their Affinities for Heavy Metal Ions / 微生物学通报

Selection and affinities of Cd binding peptides were assayed by phage random dodecapeptide library and affinity chromatography. Two Cd binding peptides were obtained, it was found that the affinities of Cu~ 2+ ,Co~ 2+ ,Zn~ 2+ ,Ni~ 2+ for Cd binding peptides were higher than that of Cd~ 2+ and Cr~ 2+ after detection of the amplified Cd binding peptides displayed phages to different heavy metal-charged resins; the detoxification of E.coli to Ni~ 2+ and Cd~ 2+ was enhanced when infected by Cd binding peptide displayed phages as compared with those of the control. The interaction of Cd binding peptide displayed phages with heavy metals charged resins was also observed under microscope. The work would be of great value and consequences for the study of interaction between heavy metals and proteins(peptides), as well as thedetoxification and bioremediation of heavy metals.