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 identify pathological mutation of D4Z4 in a child with facioscapulohumeral muscular dystrophy (FSHD) presented initially as mental retardation.Methods Wechsler Intelligence Scale for Children Revised in China (WISC-Ⅳ) was used to assess the patient's IQ.Other clinical data was also collected.With genomic DNA extracted from peripheral blood samples,the child and his parents were subjected to medical exome sequencing and copy number variation analysis by next generation sequencing (NGS).The D4Z4 repeats and their origin source were determined by molecular combing.Results By the WISC-Ⅳ test,the child was found to have a total IQ of 41,with a speech comprehension IQ of 45,and perceptual inference index IQ of 52.No pathological mutation was detected by NGS.By molecular combing method,the child was found to carry a D4Z4 spanning 5.2 kb with a copy number of 2.Analysis of his parents indicate that the mutation was de novo.Conclusion The D4Z4 copy number variation may account for the FSHD and mental retardation in the child.The molecular combing method can be used to identify the number of repeat units and facilitate the diagnosis of FSHD.

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 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.

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.

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.

Objective To explore the clinical characteristics and treatment of a family with inherited generalized epilepsy with febrile seizures plus (GEFS+) caused by STX1B gene mutation and to review the literature.Methods Clinical data of a child with GEFS+ and his family members who visited Guangzhou Women and Children Medical Center in August 2017 were collected.DNA samples of the proband and his parents,his grandparents were analyzed by the next-generation sequencing and confirmed by Sanger sequencing.A literature search with "STX1B " as the key word was conducted at PubMed,China National Knowledge Infrastructure and Wanfang databases to include recently published studies (up to May 2018).Results The proband was a 2-year-old boy who was admitted to our hospital because of frequent epileptic seizures in a month,with focal seizures evolving into the generalized bilateral tonic-clonic epileptic attacks after febrile seizure.His seizures were well controlled by levetiracetam.His father presented febrile seizure in early childhood stage and epilepsy in adult stage,which were under control with lamotrigine.The proband's grandmother had a history of febrile seizure.Other family members had no history of convulsion.Ictal electroencephalogram showed low voltage fast activities (8-10 Hz) originating from the central region of the brain.Heterozygous mutation of STX1B gene (c.705C＞G,p.Asn235Lys) was detected in the proband,his father and grandmother.The mutation has not been reported previously.Furthermore,no other family members carried the mutation at c.705 locus of STX1B gene.No article in Chinese was found,and 4 articles in a language other than Chinese provided the complete case data of 9 mutation loci in 33 patients (4 families and 4 sporadic cases).With this research data (3 cases in 1 family) included,there were 36 cases and 10 mutation sites in STX1B gene.Of these mutations,there were 5 missense mutations,3 nonsense mutations,1 insert mutation and 1 deletion mutation;and according to the mutation distribution,there were 4 mutation sites in exon 3,2 in exon 8,2 in exon 9,and 1 in exon 1.There was incomplete penetrance in the family,so different types of epileptic attacks occurred among different family members.Conclusions In this study,a de novo mutation of STX1B gene in a family with GEFS+ was defined,which would expand the gene mutation spectrum and provide basis for family genetic counseling.Clinical heterogeneity was found in this family.Seizures caused by STX 1B gene mutation were sensitive to antiepileptic drugs.

Objective:To understand the agglutination characteristics of different subtypes of avian influenza viruses, we selected erythrocytes from different sources to find suitable erythrocytes for influenza environmental sample detection.Methods:Different subtypes of avian influenza viruses, which were isolated from environmental sample between 2009 and 2016 in China, were selected to do hemagglutination assay using 5 animal erythrocytes (chicken, turkey, guinea pig, horse, and sheep). Flow cytometry was used to detect expression level and type of sialic acid receptors of different erythrocytes, and the characteristics of the receptor binding domain (RBD) of the viral hemagglutinin protein were analyzed by amino acid sequence.Results:In this study, a total of 28 strains of avian influenza virus including 14 subtypes were detected. The result showed that all viruses could agglutinate with turkey and guinea pig erythrocytes and the rest three erythrocytes were unable to produce agglutination with some viruses; among them, one H9N2 virus (A/environment/Anhui/43762/2015) did not agglutinate with chicken erythrocytes, one H1N1 virus (A/environment/Shandong/76972/2014) and two H9N2 viruses (A/environment/Chongqing/79449/2014 and A/environment/Anhui/43762/2015) did not agglutinate with horse erythrocytes, two viruses of H9N2 (A/environment/Chongqing/79449/2014 and A/environment/Anhui/43762/2015) and two viruses of H13N8 (A/environment/Qinghai Lake/166/2012 and A/environment/Qinghai Lake/13/2012) did not agglutinate with sheep erythrocytes. The result of flow cytometry showed that two sialic acid receptors, α-2, 3 and α-2, 6, were detected on the surface of erythrocytes of turkey, chicken and guinea pig, but the expression ratios of the two receptors were different. Only the expression of α-2, 3 sialic acid receptors was detected in horse and sheep erythrocytes. Sequence analysis suggested that amino acid substitution in key regions of viral hemagglutinin protein RBD may be an important factor affecting the binding properties of different erythrocytes.Conclusions:Our result suggested that turkey and guinea pig erythrocytes are the most sensitive in the hemagglutination test. Receptor expression and type of erythrocytes from different sources can significantly affect the agglutination reaction of different subtypes of avian influenza virus, and the amino acid changes in key regions of RBD can also affect the result of agglutination reaction.