Objective To investigate the genotype in Chinese patients with nonclassical 21 hydroxylase deficiency (NC 21OHD). Methods Eight patients with NC 21 OHD, 35 patients with classical one and 20 normal controls were studied as followed: CYP21 gene was amplified into fragment 1 (exon1→exon3) and fragment 2 (exon3→exon10) through PCR with specific primers. Second round PCRs were performed using fragment 1 and 2 as template, and PCR products were digested by restrictive endonucleases to analyze mutations by 3% 4% agarose gel electrophoresis. Results (1) The most frequent mutation in 8 cases of Chinese NC 21 OHD was P30L(6/16,37.5%), followed by V281L(4/16,25.0%). NC 21 OHD also carried mutations causing moderate or severe degree of enzymatic compromise, such as i2g (3/16, 18.8%), Q318X and R356W (1/16, 6.3% respectively), and I172N (3/16, 18.8%). (2) In ACTH 1 24 stimulation tests of 8 NC 21 OHD patients, basal 17 OH progesterone level was (23.9?28.4)?g/L and stimulated 17 OH progesterone level was (92.0?83.7)?g/L. (3) Genotype had strong correlation with phenotype in 21 OHD patients. Conclusion (1) As compared with caucasians, the most frequent mutation in Chinese NC 21 OHD is P30L, not V281L. (2) Much more attention should be paid to patients with hyperandrogenism to screen NC 21 OHD.

Objective To analyze the clinical features and risk factors of invasive fungal infections (IFI) in children with acute leukemia.Methods Ninety-six acute leukemia children complicated with IFI admitted in Guangzhou Women and Children's Medical Center during January 2005 and February 2017 were retrospectively reviewed, and 96 cases of acute leukemia without IFI admitted at the same period were randomly selected as control group.The clinical manifestations of IFI were analyzed, multivariate Logistic regression was used to study risk factors of the complication of IFI in pediatric acute leukemia.Results Among 96 children complicated with IFI, fungus were detected in samples from sputum, bronchoalveolar lavage fluid, or blood in 78 cases, in which 42 cases (43.75％) were oral infection, 36 cases (37.50％) were pulmonary infection.Candida albicans (33.33％, 26/78) was the most commonly isolated pathogen, followed by Candida parapsilosis (20.51％, 16/78) and Candida tropicalis (20.51％, 16/78).Univariate analysis revealed hormone-containing chemotherapy, neutropenia (＜ 0.5 × 109/L), time duration of neutropenia ≥ 10 days, usage of carbapenem antibiotics and combined drug administration ≥2 types were associated with fungal infection (P ＜ 0.05 or ＜0.01).Multivariate Logistic regression showed that the time duration of neutropenia ≥ 10 days (OR =11.390, 95％ CI 4.145-55.263, P ＜ 0.01),usage of carbapenem antibiotics (OR =4.825, 95％ CI 1.681-13.842, P ＜ 0.01) and hormone-containing chemotherapy (OR =2.220, 95％ CI 1.542-8.246, P ＜ 0.05) were the independent risk factors of IFI.Conclusion Rational usage of antibiotics and effective measures taken to restore the granulocytes can help to reduce the incidence of IFI in children with acute leukemia.

25 years group. Among 129 cases who had taken antihypertensive drugs before operation, only 43 cases continued to use drugs after operation, therefore, rate of antihypertensive drugs administration decreased significantly after operation (P

Aged ; Antineoplastic Agents ; therapeutic use ; Benzamides ; therapeutic use ; Drug Resistance, Neoplasm ; Exons ; Gastrectomy ; methods ; Gastrointestinal Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Gastrointestinal Stromal Tumors ; drug therapy ; metabolism ; pathology ; surgery ; Humans ; Imatinib Mesylate ; Liver Neoplasms ; drug therapy ; secondary ; Male ; Piperazines ; therapeutic use ; Point Mutation ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Pyrimidines ; therapeutic use

Objective: To acquire recombinant nematode anticoagulant peptide ( NAP) with high anticoagulant activity. Methods: Pichia pastoris GS115 strain was transformed with recombinant yeast expression vector pPICS. 5K-rNAP. Expression of rNAP was induced with methanol after the identification of positive strains. NAP expressed in the collected yeast culture supernatant was confirmed with SDS-PAGE and Western blot. The biological activity of the products was validated with PT (prothrombin time) , INR (international normalized ratio) and APTT (activated partial thromboplastin time) , respectively. Results: The yeast strains expressing NAP were identified. The rNAP was secreted into culture supernatant with a molecular weight of about 10 kDa due to glycosylation, which is a little bigger than that predicted (8.7kDa). The anticoagulant efficiency of rNAP was confirmed with the in vitro assays. Conclusion: The recombinant nematode anticoagulant peptide with high biological activity was successfully expressed in Pichia pastoris and can be used in the future development of novel anticoagulant agent.

Objective: To construct a polycistron tandem repeated Echistatin (Ecs) gene. Methods: Three Ecs genes with independent initiation and termination codon were ligated tandem through restriction enzyme sites after amplified with 3 pairs of primers using pMD18T-Ecs as template. The polycistron Ecs gene was inserted into pET30a and expressed in E.coli BL21(DE3) with IPTG induction. The expression results were identified by 18% SDS-PAGE and Western blot. Results: The expression of Ecs polycistron was accomplished with 18% expression level of total protein determined by SDS-PAGE and Western blot. Conclusion: The successful expression of Ecs polycistron provided a new method for the preparation of low molecular weight protein.

Objective To analyze the clinical characteristics of urinary tract infection after kidney transplantation and to analyze the distribution of pathogenic virulence factors.Methods From January 2014 to June 2016,a total of 255 cases were subjected to DCD renal transplantation in our center,and 45 cases of urinary tract infection occurred after operation.Among them,urinary tract E.coli infection occurred in 32 cases.The virulence factors gene aer,hly,irp2 and iucD in the 32 strains of E.coli isolated from urine were tested using PCR and multiple PCR.Results Thirty-two cases (71.1％) of urinary tract infection after renal transplantation was caused by E.coli,and divided into 9 cases of nosocomial infection group,and 23 cases of community-acquired group.The positive rate for aer,hly,irp2 and iucD in Nosocomial infection group was 66.7％,33.3％,88.9％ and 77.8％,and that in community-acquired group was 30.4％,8.70％,39.1％ and 34.7％,respectively.There were statistically significant differences among four kinds of virulence factors between two groups (P＜0.05).The positive rate of virulence factors was significantly higher in nosocomial infection group than in community-acquired groups.The total resistance rate was 84.38％,and there was significarit difference in multi-drug resistance rate between two groups (P＜0.01).Conclusion E.coli is the main pathogenic bacteria of urinary tract infection after renal transplantation,especially nosocomial infections.The detection rate of virulence factor is high,and the multiple drug resistance is outstanding,so the attention should be paid clinically.

Objective To explore the peripheral blood T lymphocytes cell level in different grades of cerebral gliomas pa-tients at the time of before and after operation and its clinical significance .Methods A total of 80 cases of brain tumor patients from February 2010 to February 2012 in this hospital were chosed as study objects ,included 57 cases of glioma ,23 cases of other brain tumors ;57 cases of glioma were divided into 19 cases of low grade group and 38 cases of high grade group accorded to WHO grading standards ,and 50 cases of healthy people in same period were selected as control group .Venous blood in three groups were extracted at 1st day and 1st week after operation ,to detected the level of T lymphocyte subsets in peripheral blood and analyze the relationship between the different grade gliomas and prognosis .Results The peripheral blood CD3 + ,CD4 + ,CD4 + /CD8 + in control group were significantly higher than that of other brain tumor group and the glioma group ,CD8 + was significantly lower than that of the two groups(P< 0 .05) .Compared with other tumor group ,peripheral blood CD3 + ,CD4 + ,CD4 + /CD8 + in glioma group de-creased significantly ,CD8 + increased significantly(P< 0 .05) ;before and after the operation ,the CD3 + and CD4 + /CD8 + levels were significantly higher in low grade group than in the high grade group ,CD8 + was significantly lower in low grade group than in the high grade group(P< 0 .05) ,at the time of 1st week after operation ,CD3 + and CD4 + /CD8 + increased ,CD8 + decreased significant-ly of two groups when intra-group comparison (P < 0 .05) .The median survival time were 31 months in low grade group and 13 months in high grade group ,and the singnificance was found in two groups(P< 0 .05) .The median survival time were 34 months in CD4 + /CD8 + > 1 group and 17 months in CD4 + /CD8 + < 1 group ,the singnificance was found in two groups(P< 0 .05) .Conclusion The immune function of patients with brain glioma is inhibited ,the higher the malignancy of the tumor ,the more obvious of im-mune inhibition ;T lymphocyte subsets level could be used as evaluating index of malignant degree and prognosis of brain glioma .

Objective To explore a simpler, more economic and reproducible method to reproduce a model of high oxygen induced acute lung injury (HALI) in rats. Methods An animal feeding box equipped with a controllable high oxygen was designed. 100 Sprague-Dawley (SD) rats were divided into normal control group and HALI group by random number table method, with 50 rats in each group. Each group was randomly subdivided into five subgroups according to the duration of exposure to high oxygen, namely 0, 24, 48, 72 and 96-hour subgroups, with 10 rats in each subgroup. The rats in normal control group were kept in cages with ambient air, and the rats in HALI group were kept in an oxygen tank in which the oxygen concentration was higher than 90% volume ratio, with the temperature maintained at 25-27 ℃, humidity of 50%-70%, and CO2 concentration < 0.5% for 23.5 hours every day. The arterial blood of rats was collected for analysis of blood gas at all time points, and the oxygenation index (OI) and respiratory index (RI) were calculated. Then the rats were sacrificed and the right lung was harvested, which was sectioned and stained with hematoxylin and eosin (HE). The changes in histopathology were observed with light microscopy, and pathological score was recorded. The left lung was harvested for the measurement of the wet/dry weight ratio (W/D). Results With the prolongation of high oxygen exposure time, the degree of lung injury in HALI group was gradually increased, and the degree of derangement of alveolar structure appeared in an increasing degree, with destruction of the alveolar wall, widening of alveolar space, and appearance of edema, and inflammatory cell infiltration. A small quantity of red blood cells exudation could be found in some rats. The pathologic changes were most obvious at 48-72 hours after exposure. With the prolongation of high oxygen exposure time (0, 24, 48, 72, 96 hours), the OI (mmHg, 1 mmHg = 0.133 kPa) in HALI group was gradually decreased (446.67±29.93, 306.19±37.23, 269.70±29.00, 253.81±43.40 and 245.58±35.25), RI, pathological score of lung tissue and W/D ratio were gradually increased [RI: 0.25±0.04, 0.31±0.06, 0.38±0.06, 0.46±0.07 and 0.44±0.03; pathological score of lung tissue: 0.00±0.00, 0.90±0.74, 2.90±1.20, 4.70±1.57 and 4.80±1.23; lung W/D ratio: 3.84±0.61, 4.14±0.46, 4.56±0.34, 5.32±0.27 and 5.18±0.25]. Statistically significant differences were found in 72-hour group as compared with that of other groups (all P < 0.05), while no significant difference was found between 96 hours and 72 hours groups (all P > 0.05). There were significant differences in changes between 24, 48, 72, and 96 hours as compared with those of the normal control group: OI (mmHg): 24 h 306.19±37.23 vs. 435.65±25.34 and 96 h 245.58±35.25 vs. 465.42±24.75; RI: 24 h 0.31±0.06 vs. 0.24±0.04 and 96 h 0.44±0.03 vs. 0.24±0.06. The same as true in pathological scores of lung tissue: 24 h 0.90±0.74 vs. 0.00±0.00 and 96 h 4.80±1.23 vs. 0.00±0.00; lung W/D ratio: 24 h 4.14±0.46 vs. 3.79±0.44 and 96 h 5.18±0.25 vs. 4.12±0.91, all P < 0.05. Conclusions A self-designed high oxygen box is simple, easy to operate and reproduction of HALI model can be attained. Sustained exposure to high concentrations of oxygen (≥ 90%) for 24 hours can replicate the HALI model successfully, and the most serious injury appears at 48-72 hours after exposure.

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