Objective To find effective measures to reduce blood transfusion reactions through retrospective analysis on adverse reactions of blood transfusion .Methods Statistically analyse the the occurrence of adverse transfusion reactions from Jan .2011 to May 2015 .Results The total adverse transfusion reaction rate was 0 .31% (136/43 874) .The adverse reaction rate of cryoprecipi‐tate infusion was the highest(1 .27% ) in all kinds of blood components infusions ,followed by whole blood(0 .56 % ) ,red blood cell suspension(0 .44% ) ,frozen plasma (0 .32% ) ,platelets (0 .27% ) ,leukocyte -depletion red blood cell suspension (0 .00% ) and washed red blood cell(0 .00% ) .When considering the times of blood transfusion ,the reaction rates of once ,twice ,3 times and ≥4 times were 0 .19% ,0 .27% ,0 .66% and 1 .24% respectively .The incidences of adverse reactions caused by different blood transfu‐sion were statistically significant(P< 0 .05) .60 .29% adverse transfusion reactions happened to people with a history of blood transfusion .The most common adverse transfusion reaction was non-hemolytic reaction which accounted for 62 .50% ,followed by allergic reactions accounting for 33 .09% .Conclusion Leukocyte-depletion red blood cells suspension is proved to be safer ,and could reduce the transfusion reactions .Leukocyte-depletion red cell suspension could benefit the patient who needs blood transfu‐sion more than twice .

Objective To explore the killing effect of dielectric barrier discharge (DBD) plasma on tumor cells and to analyze the DBD-induced apoptosis mechanism.Methods Thiazolyl blue tetrazolium bromide (MTT) assay method was used to detect the killing effect of low temperature plasma on the cytotoxicity of normal spleen leukocytes and acute promyelocytic leukemia cells (LT-12) at different doses.The changes of reactive oxygen species (ROS) level were measured after plasma treatment.The cell apoptosis rate was detected by Annexin V/PI double staining at different doses.The expression of apoptosis-related genes and proteins was detected by qRT-PCR and Western Blot.Results MTT results showed that the killing effect of plasma treatment was dose-dependent and time-dependent.The cell survival rate after 8 hours of treatment decreased from 98％ to 63％ with the dose increasing from 30 s to 240 s.The survival rate decreased from 78％ (2 h) to 39％ (24 h) after the treatment with a same dose (e.g.240 s).Annexin V/PI double staining results demonstrated that the plasma effect can induce apoptosis,and the apoptosis rate was not only positively correlated with the plasma dose,but also with the post-plasma time.The longer the post-plasma time,the higher was the apoptosis rate.The apoptotic rate of the 60 s dose treatment after 12 h was 48％ that increased to 55.3％ with the dose of 120 s.The production of reactive oxygen species (ROS) detected by flow cytometry also showed a time correlation of the plasma treatment.After the plasma treatment,the ROS level immediately increased to 1.24 times,and sharply increased to 5.39 times after 20 h post-plasma.The experimental results of qRT-PCR and Western blot showed that the expression of the genes and proteins of Caspase family and Bcl-2 family was very active at 8 to 12 h post-plasma treatment.Conclusions Low-temperature plasma can effectively kill tumor cells,and apoptosis is the main mechanism of death.The molecular mechanism of apoptosis of tumor cells induced by low temperature plasma was preliminary confirmed.

OBJECTIVETo develop a hydroponic Nicotiana cultivation system for rapid and high-yield transient expression of recombinant proteins under laboratory conditions.

METHODSTo establish the hydroponic cultivation system, several parameters were examined to define the optimal conditions for the expression of recombinant proteins in plants. We used the green fluorescent protein (GFP) and the geminiviral plant transient expression vector as the model protein/expression vector. We examined the impact of Nicotiana species, the density and time of Agrobacterium infiltration, and the post-infiltration growth period on the accumulation of GFP. The expression levels of GFP in Nicotiana leaves were then examined by Western blotting and ELISA.

RESULTSOur data indicated that a hydroponic Nicotiana cultivation system with a light intensity of 9000 LX/layer, a light cycle of 16 h day/8 h night, a temperature regime of 28 degrees celsius; day/21 degrees celsius; night, and a relative humidity of 80% could support the optimal plant growth and protein expression. After agroinfiltration with pBYGFPDsRed.R/LBA4404, high levels of GFP expression were observed in both N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants cultured with this hydroponic cultivation system. An optimal GFP expression was achieved in both Nicotiana species leaves 4 days after infiltration by Agrobacterium with an OD(600) of 0.8. At a given time point, the average biomass of N. tobaccum (cv. Yuyan No.5) was significantly higher than that of N. benthamiana. The leaves from 6-week-old N. benthamiana plants and 5-week-old N. tobaccum (cv. Yuyan No.5) plants could be the optimal material for agroinfiltration.

CONCLUSIONWe have established a hydroponic cultivation system that allows robust growth of N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants and the optimal GFP expression in the artificial climate box.

Gene Expression Regulation, Plant ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Hydroponics ; methods ; Plants, Genetically Modified ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Tobacco ; genetics ; growth & development

Objective To investigate the relationship between vitamin D level and thyroid peroxidase (TPO) level in Qinghai Province.Methods In 2015 and 2016,a stratified cluster sampling method was used to select 1 873 people in 9 regions of Qinghai Province,including 935 males and 938 females,the average age was (52.3 ± 10.9) years old,there were 1 040 Han people and 833 Tibetans.Fasting venous blood samples were collected,and vitamin D and TPO levels were detected by a fully automated chemiluminescence immunoassay system,vitamin D and TPO levels were compared in different gender,ethnic group,and altitude populations.Results The vitamin D level was (16.3 ± 7.5) μg/L,and abnormal rate of vitamin D was 89.2％ (1 670/1 873);the TPO level [median (quartile)] was 23.8 (16.0-35.0) U/ml,and the positive rate of TPO was 26.8％ (502/1 873).The vitamin D levels of males and females were (16.9 ± 7.6) and (15.6 ± 7.6) μg/L,respectively,the difference was statistically significant (t =3.684,P ＜ 0.01);the TPO levels were 23.8 (16.4-31.0) and 23.8 (15.9-37.6) U/ml,respectively,the difference was not statistically significant (Z =-1.084,P ＞ 0.05).The vitamin D levels of the Han people and the Tibetans were (16.2 ± 7.8) and (16.3 ± 7.3) μg/L,respectively,the difference was not statistically significant (t =-0.110,P ＞ 0.05);the TPO levels were 23.5 (15.7-34.8) and 24.0 (16.5-35.1) U/ml,respectively,the difference was not statistically significant (Z =-0.078,P ＞ 0.05).The vitamin D levels of middle-altitude (1 500-3 000 m) and high-altitude (＞ 3 000 m) populations were (16.6 ± 7.7) and (15.8 ± 7.5) μg/L,respectively,and the difference was not statistically significant (t =2.126,P ＞ 0.05);the TPO levels were 23.4 (16.0-33.0) and 24.0 (16.0-36.8) U/ml,respectively,the difference was not statistically significant (Z =-1.296,P ＞ 0.05).There was negative correlations between vitamin D level and TPO level,altitude (r =-0.150,-0.052,P＜ 0.05),and no correlation with ethnic group (r =0.003,P ＞ 0.05).Conclusions The vitamin D level is negatively correlated with TPO level in Qinghai population.The abnormal rate of vitamin D is high,and vitamin D level of males is higher than that of females.

Objective To investigate the iodine nutritional and thyroid stimulating hormone level of patients with thyroid nodules in different regions of Qinghai Province and analyze the characteristics of changes in different regions.Methods In 2014-2016,thyroid nodules in 9 regions of Qinghai Province (Tibetan areas:Xiewu,Nangqian,Jiegu,Guoluo;non-Tibetan areas:Xining,Huzhu,Menyuan,Minhe,and Ledu) were selected and serum thyroid stimulating hormone (TSH) and urine iodine were measured.Results A total of 553 thyroid nodules,the median urinary iodine (MUIC) was 160.8 μg/L and the median TSH was 2.97 mU/L.The iodine nutritional status was at an appropriate level.Among them,MUIC (206.8 μg/L) in thyroid nodules in the Menyuan area was slightly higher than the appropriate amount,there was a significant difference in MUIC among different region (x2 =47.747,P ＜ 0.05);of TSH in thyroid nudules in the 9 regions,the differences were statistically significant (x2 =34.832,P ＜ 0.05).Non-Tibetan areas were compared with Tibetan areas,there was a significant difference in MUIC (155.6,185.6 μg/L),TSH (2.68,3.45 mU/L,Z =-3.677,-5.410,P ＜ 0.05);Among them,the differences was statistically significant between MUIC (152.8,187.7 μg/L) of women with thyroid nodules (Z =-3.504,P ＜ 0.05);there was a statistically significant difference in TSH levels among men (2.58,3.46 mU/L) and women (2.80,3.44 mU/L) with thyroid nodules (Z =-3.613,-4.040,P ＜ 0.05);there were no significant differences in MUIC levels among thyroid nodules of each age groups (P ＞ 0.05);of the TSH level in 30-and 50-＜ 65 years groups (2.63,3.17;2.25,3.58 mU/L),the differences were statistically significant (Z =-2.892,-3.233,P ＜ 0.05),and other groups were no significant differences (P ＞ 0.05).Conclusion The iodine nutrition of patients with thyroid nodules in these regions of Qinghai Province is generally at an appropriate level,the MUIC and TSH levels in Tibetan areas were lower than those in non-Tibetan areas,and iodine nutrition status and TSH levels should be monitored for key populations.

Objective:To summarize the clinical phenotype and CUX2 gene variation characteristics of developmental epileptic encephalopathy type 67 confirmed by whole exome sequencing. Methods:Clinical data of 1 case diagnosed as CUX2 gene mutations related developmental epileptic encephalopathy type 67 in the Children′s Hospital Affiliated to Zhengzhou University in January 2021 were collected, the patient′s clinical characteristics, genetic testing, head imaging, electroencephalogram results and treatment were summarized, and the patient was regularly followed-up every 3 months. At the same time, the domestic and foreign literatures on epileptic encephalopathy caused by CUX2 gene mutation were reviewed. Results:The proband was a 6 years and 4 months old girl. The main clinical manifestations included focal origin progression to bilateral tonic-clonic seizures, retardation of intellectual, language, and motor development, autistic behavior, hyperactivity disorder, and involuntary hand clapping. The video electroencephalogram showed extensive spiny slow wave and multi-spiny slow wave emission in waking and sleeping stages, and spiny slow wave and spiky slow wave emission in bilateral anterior head in sleeping stage. Brain magnetic resonance imaging (MRI) plain scan and T 2-fluid attenuated inversion recovery (T 2-FLAIR) thin layer scan showed that the signal of the left hippocampus was higher than that of the right, and the left hippocampus was slightly swollen. One month later, the brain MRI and T 2-FLAIR were reexamined. The left hippocampal signal was still slightly higher and decreased, and the hippocampal volume was slightly reduced. Whole exome sequencing showed the CUX2 gene with c.1768G>A(p.Glu590Lys) heterozygous missense variant, which was a reported de novo pathogenic variant and both of her parents were wild-type. A total of 10 cases of new heterozygous missense variants in CUX2 gene [c.1768G>A (p.Gelu590Lys)] were reported in 4 literatures. No relevant cases have been reported in China. Conclusions:Developmental epileptic encephalopathy type 67 is relatively rare. The main clinical features are seizures, global developmental delay, movement disorders, athetosis, autism and hyperactivity disorder. The heterozygous missense variant c.1768G>A (Glu590Lys) of CUX2 gene maybe the genetic cause of this case.

Clinical data and genetic mutation characteristics of a patient with Coffin-Siris syndrome by 6q25.3 deletion were summarized. The child was a 7-year and 6-month old girl who had feeding difficulties, repeated infection, language and motor retardation, low intelligence, laryngeal cartilage dysplasia, thick eyebrows, sparse teeth, hairy back, hyperactivity and aggressive behavior, seizures and ataxia. There was no abnormality in chromosomal karyotype analysis by proband; genomic copy number variant sequencing (CNV-seq) indicated approximately 4.27 Mb heterozygous deletion in chromosome 6q25.3 region, with 17 genes including ARID1B gene, father maternal CNV-seq showing no abnormalities. Trio-whole-exome sequencing showed the proband missed all exons 1-20 of the ARID1B gene, with wild-type parents. The proband had severe clinical symptoms and haplodose insufficiency which was the genetic etiology.