Construction and expression of circular RNA in vitro / 中国生物制品学杂志

@#Objective To construct encoding RNA that can be cyclized in vitro by using the permuted intron exon(PIE)strategy in the maturation process of eukaryotic mRNA,and transfect it into HEK-293T cells for expression.Methods The sequences of 5'and 3'cyclic arms with groupⅠcatalytic intron,the internal ribosome entry sites(IRES)of Coxsackievirus B3(CVB3)and the target gene were selected to construct the template plasmid. Linearization plasmid template obtained by PCR was used to synthesize linear RNA through in vitro transcription(IVT),which then started in vitro cyclization(IVC)by the addition of cyclization reagents to obtain circular RNA(circRNA). RNA cyclization was confirmed by agarose gel electrophoresis and ribonuclease R(RNase R)digestion. HEK-293T cells were transfected with circRNAs respectively carrying enhanced green fluorescent protein(EGFP),firefly luciferase(Fluc),and influenza virus hemagglutinin(HA)IVR-180 genes,to verify their expression with in vitro.Results With RNA cyclization,the main band of agarose gel electrophoresis became smaller and small fragments appeared. After RNase R digestion,only some circRNA bands remained.HEK-293T cells transfected with EGFP-circRNA showed significant green fluorescence under the fluorescence microscope.The Fluc expression values of HEK-293T cells transfected with Fluc-circRNA were on average 20 times higher than non cyclized RNA,and the relative light unit(RLU)scaled up with the increase of Fluc-circRNA transfection dose. Western blot analysis showed that HA protein was successfully expressed in HEK-293T cells transfected with HA-circRNA.Conclusion In this study,linear RNA was successfully cyclized in vitro and different proteins were expressed,which lays a foundation of the research of new influenza vaccines and mRNA vaccines.

CD44-HA Mediate Adhesion of MDA-MB-231 Cells and HL60 Cells under Fluid Shear Flow / 医用生物力学

Objective To explore how hyaluronic acid ( HA) in extracellular matrix regulates the adhesion ofCD44+tumor cells. Methods MDA-MB-231 cells or HL60 cells were perfused in a parallel plate chamber. Themovement of cells over immobilized HA was observed and analyzed to obtain the characteristics of cell adhesionand rolling. Results The adhesion number of MDA-MB-231 cells on HA substrate was positively regulated by HAconcentration, but not by HA molecular weight. Compared with physically adsorbed HA, immobilized HA byavidin-biotin could significantly improve the cell adhesion ratio. With the increase of shear stress in the range of30-50 mPa, the rolling velocity of cells increased and the adhesion ratio decreased, but the tether lifetime of cellswas not affected. In the same flow field, compared with MDA-MB-231 cells, HL60 cells with low expression ofCD44 rolled more quickly on immobilized HA, with shorter tether lifetime and much lower adhesion ratio(<1. 5% ). Conclusions Fluid shear stress might mediate the rolling velocity of MDA-MB-231 cells by regulatingthe CD44-HA association rate rather than their dissociation rate. The interaction between CD44 and HA is involved in the initial adhesion of HL60 cells, but it does not play a major role. This study will provide references for the design of anti-tumor drugs.

Prokaryotic expression, polyclonal antibody preparation, spatio-temporal expression profile and functional analysis of c-Myc of Helicoverpa armigera (Lepidoptera: Noctuidae) / 生物工程学报

c-Myc protein encoded by c-Myc (cellular-myelocytomatosis viral oncogene) gene regulates the related gene expression through the Wnt/β-catenin signaling pathway, and has received extensive attention in recent years. The purpose of this study was to express Helicoverpa armigera c-Myc gene (Ha-c-Myc) by using prokaryotic expression system, prepare the polyclonal antibody, examine the spatio-temporal expression profile of Ha-c-Myc, and investigate the possible function of Ha-c-Myc in regulating H. armigera sterol carrier protein-2 (SCP-2) gene expression. The Ha-c-Myc gene was amplified by PCR and cloned into a prokaryotic expression plasmid pET-32a(+). The recombinant plasmid pET-32a-Ha-c-Myc was transformed into Escherichia coli BL21. IPTG was used to induce the expression of the recombinant protein. Protein was purified by Ni2+-NTA column and used to immunize New Zealand rabbits for preparing the polyclonal antibody. The Ha-c-Myc expression levels in different developmental stages (egg, larva, prepupa, pupa, and adult) of H. armigera and different tissues (midgut, fat body, head, and epidermis) of the prepupa were determined by real-time quantitative reverse transcription PCR (qRT-PCR). Ha-c-Myc siRNA was synthesized and transfected into H. armigera Ha cells. The relative mRNA levels of Ha-c-Myc and HaSCP-2 in Ha cells were detected by qRT-PCR. Results showed that the pET-32a-Ha-c-Myc recombinant plasmid was constructed. The soluble Ha-c-Myc protein of about 65 kDa was expressed in E. coli. The polyclonal antibody was prepared. Western blotting analysis suggested that the antibody had high specificity. Enzyme linked immunosorbent assay (ELISA) showed that the titer of the antibody was high. Ha-c-Myc gene expressed at all developmental stages, with high levels in the early and late instars of larva, and the prepupal stage. Tissue expression profiles revealed that Ha-c-Myc expressed in various tissues of prepupa, with high expression level in the midgut, but low levels in the epidermis and fat body. RNAi results showed that the knockdown of Ha-c-Myc expression significantly affected transcription of HaSCP-2, leading to a 50% reduction in HaSCP-2 mRNA expression level. In conclusion, the Ha-c-Myc was expressed through a prokaryotic expression system, and the polyclonal anti-Ha-c-Myc antibody was obtained. Ha-c-Myc may promote the expression of HaSCP-2 and play an important role in the lipid metabolism of H. armigera. These results may facilitate further study on the potential role and function mechanism of Ha-c-Myc in H. armigera and provide experimental data for exploring new targets of green pesticides.

Nutritional assessment of servicemen in relation to area of duty and feeding habits: a Pakistani prospective

Abstract The present study was carried out to determine incidence of overweight and obesity in Pakistani servicemen with reference to their area of duty, feeding habits and also to identify risk factors. Accordingly, 2,501 servicemen selected from all over Pakistan using multiple stage stratified sampling protocol. Nutrition assessment performed using body mass index (BMI), waist to hip ratio (WHR) and dietary assessment using food frequency questionnaire. Collected data was analyzed using the SPSS version 25. Regression was used to find risk factors of obesity and WHR. Results indicated that about 1/4th of servicemen were smokers. Approximately, 1/5th of them were overweight and about one quarter were eating fruits and vegetables for 3 days/ week and 4 days/week, respectively. Only 1/3rd of them were physically active for at least 40 minutes per day. Age and fruits intake were significantly predicting BMI with a direct relation and vegetable intake was negatively correlated to BMI of the servicemen. Age and rank were significant predictors of WHR while, physical activity was negatively correlated to WHR. It is concluded and suggested from our study that there is a need to modify eating patterns and habits as well as improving physical activity on daily basis for healthy and long life of the servicemen.

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Sequence analysis of HA and NA genes of human infected H9N2 avian influenza virus in Yunnan province, 2019 / 中国热带医学

@#Abstract: Objective To investigate the molecular characteristics of the H9N2 avian influenza virus (AIV) causing human infection in Yunnan Province in 2019, and to provide the scientific basis for the prevention and control of avian influenza in Yunnan Province. Methods Influenza virus typing was performed by real-time RT-PCR in two influenza-like illness samples, and the Illumina Miseq high-throughput sequencer was used to determine the viral genome sequence. HA and NA gene sequence alignment and phylogenetic tree construction were performed using Mega7.0 software. Results Real-time RT-PCR results showed that two influenza-like illness samples were positive for H9N2 subtype. The full length of HA and NA were obtained by genomic sequencing. Sequence system evolution analysis showed that the HA and NA of the two AIVs in Yunnan Province were in the same evolutionary clade as A/Chicken/Zhejiang/HJ/2007 and belonged to the G57 type. The HA nucleotide and amino acid homology of the two AIVs were 93.92% and 95.00%, respectively, and the NA nucleotide and amino acid homology was 93.31% and 82.03%, respectively. The nucleotide (amino acid) homology of HA was 92.29%-96.94% (93.77%-98.43%) and 92.84%-94.92% (94.18%-96.23%), respectively, and NA nucleotide homology (amino acid) were 91.81%-97.60% (77.82%-94.83%), 94.38%-97.22% (85.47%-94.55%), respectively, compared with that of human infected H9N2 epidemic strains obtained in China from 2015 to 2020. Both AIVs HA protein cleavage site sequences were PSRSSR↓GLF, which was in line with the characteristics of low pathogenic influenza. The analysis of HA protein receptor binding site showed that amino acids at positions 109, 161, 163, 191, 202, 203 and 234 were consistent with the reference strains, while amino acids at position 198 were mutated to T. N166D and 168N mutations were also found in HA protein, and both AIVs had 7 potential glycosylation sites. Analysis of the erythrocyte binding site of NA gene found that there were amino acid mutations at positions 369, 402, 403, and 432, and amino acid deletion at positions 63-65 was found in the NA genes. There were 4 and 5 potential glycosylation sites in the two AIVs, respectively, and no drug resistance site mutations were found. Conclusions　The receptor binding sites, erythrocyte binding sites and glycosylation sites of HA and NA genes of H9N2 AIV in Yunnan Province have different degrees of variation, and monitoring and prevention and control should be strengthened.

Complete genome comparative analysis of Coxsackievirus A6 causing hand,foot and mouth disease and herpangina in Yunnan Province in 2018 / 中国生物制品学杂志

@#ObjectiveTo analyze the genome-wide characteristics of 17 strains of Coxsackievirus A6(CVA6)that cause hand,foot and mouth disease(HFMD)and herpangina(HA)in Yunnan Province in 2018,and understand the genetic differences between different pathogenic CVA6.MethodsA total of 1 909 stool samples clinically diagnosed as HFMD and HA in Kunming Children′s Hospital in 2018 were randomly selected for detection using enterovirus group A universal primers and screening of CVA6 positive samples. The CVA6 whole genome sequence was amplified with CVA6 whole genome primers,spliced by BioEdit splicting software,and analyzed for the whole genome characteristics by BioEdit,MEGA 7.0,Simplot,Heml 1.0 and Phyre2softwares.ResultsA total of 929 CVA6 positive samples were screened,and 17CVA6 complete gene sequences were obtained(9 of which were clinically diagnosed as HFMD and 8 were clinically diagnosed as HA). All 17 CVA6 strains were in type IV clade on the whole phylogenetic tree. No significant recombination occurred in HA and HFMD representative strains,while mutations occurred in non-structural protein 3D region. HFMD and HA representative strains showed differences in VP1 loci S597T,Q705L and Q663L. Online predictive analysis showed that the secondary structure of VP1 was consistent with that of CVA6 with no change.ConclusionThe 17 CVA6 strains causing HFMD and HA had high genomic homology,as well as nucleotide and amino acid differences,which may affect the replication and adaptability of CVA6.

Effect of activation of RIG-Ⅰ signaling pathway by LRRC23 on replication of influenza virus / 中国生物制品学杂志

@#ObjectiveTo investigate the effect of activation of RIG-Ⅰsignaling pathway by leucine rich repeat containing23(LRRC23)on replication of influenza virus.MethodsOverexpression and knock-down of LRRC23 were performed in A549 cells to investigate its effect on influenza virus replication. A549 cells were transfected with pcLRRC23 plasmid or siLRRC23(small interfering LRRC23)for 24 h and then infected with influenza virus A/jingfang/1/86(H1N1). The virus titer and HA protein expression level in the cell supernatant were determined by plaque assay and ELISA respectively.The expression of LRRC23,RIG-Ⅰ,MAVS,M1 and HA at gene and protein levels were determined by qRT-PCR and Western blot respectively. The interactions between LRRC23 and RIG-Ⅰwere analyzed by co-immunopre-cipitation(Co-IP)and immunofluorescence assay(IFA). IFN-β-luc and NF-κB-luc activities were determined by dual-luciferase reporter assay.ResultsThe LRRC23 overexpression significantly decreased the influenza virus titer and inhibited the expression of HA protein in the supernatant of A549 cells,while enhanced the NF-κB and IFNβ activations by activation of RIG-Ⅰ-MAVS signaling pathway,resulting in the inhibition of expressions of M1 gene and HA protein. Conversely,the knock-down of LRRC23 increased the protein expression level of HA in the supernatant of A549 cells,up-regulated the relative expression level of M1 gene and down-regulated those of RIG-ⅠmRNA and MAVS mRNA.ConclusionLRRC23 plays an essential role in innate antiviral response by inhibiting influenza virus replication through activation of RIG-Ⅰsignaling pathway.

Application progress of high performance liquid chromatography in detection of hemagglutinin content in influenza vaccine / 中国生物制品学杂志

@#At present,the most commonly used method for detecting hemagglutinin(HA)content in influenza vaccines is still single-radial immunodiffusion(SRID). However,the preparation of standards required by this method takes a long time,usually 2 ～ 3 months. Therefore,how to quantitatively analyze HA accurately has always been a difficult problem in the detection of HA content in the situation that reference products can not be obtained at the early stage of the pandemic influenza. High performance liquid chromatography(HPLC)has its own characteristics of rapidity,high sensitivity,good repeatability and high accuracy,which can rapidly determine HA content by using different separation principles and has been widely used in the detection of HA content in influenza vaccine. This paper reviewed the research progress of the application of HPLC in the determination of HA content in influenza vaccine.

Outcome of Single Intra-Articular Injection of High- Molecular-Weight Hyaluronic Acid in Patients with Knee Osteoarthritis

Background: Osteoarthritis (OA) is the most common type of rheumatic disease and a leading cause of disability. Current treatments aim at alleviating these symptoms by several different methods: non-pharmacological and pharmacological treatments and invasive interventions. Among the regenerative methods, hyaluronic acid (HA) is popular now-a-days where differences exist in concentration, molecular weight, dosage, expected duration of effects and added formulations. Based on HA molecular weight, these products are classified in two groups (high and low). Due to cost effectiveness, low molecular weight HA is commonly used. To find out the outcome of single intra-articular injection of high-molecular- weight HA in patients with knee osteoarthritis.Material & Methods:This was a randomized clinical trial conducted among purposively selected 55 patients suffering from Knee OA attending at the Physical Medicine and Rehabilitation outpatient department, DMCH during July 2020 to June 2021. Patients were randomly allocated into two groups; group A received single dose of high molecular weight HA and designed exercise program and group B received same exercise program only. Pre-treatment and 3 weekly post treatment assessment were done up to 12 weeks in each group. In each follow up visit, pain, and functional status were measured in Visual Analog Scale (VAS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) respectively and compared between group A and group B where was considered as significant (p value <0.050.Results:The mean age of the participants in group A and group B were 56.8 (±9.8) and 53.3 (±7.4) years respectively where majority of the patients in both groups were female patients with overweight. In group A, VAS and WOMAC significantly decreased from base line to each follow up till 12th weeks after treatment with high-molecular-weight HA (P<0.001). In group B, VAS and WOMAC significantly decreased from base line to each follow up till 12th weeks after treatment with conservative management (P<0.001). After 6 weeks of intervention, the mean VAS scores of the patients became 4.9 ±1.1 and 6.0±1.0 in group A and group B respectively which showed significant statistical difference (P<0.001). At the end of 12 weeks, the VAS score was significantly decreased in group A (2.8±1.3) than group B (4.9±1.7) (P<0.001). After 3 weeks of intervention, the mean WOMAC scores of the patients became 71.2±6.6 and 75.0 ±4.9 in group A and group B respectively which showed significant statistical difference (P<0.001). At the end of 12 weeks, the WOMAC score was significantly decreased in group A (61.9±6.6) than group B (68.1±4.6) (P<0.001). Conclusion:Single intra-articular injection of high-molecular-weight Hyaluronic Acid is effective in patients with knee osteoarthritis in reducing pain and improving functional impairment. Long term and large scale research studies are needed to establish the effectiveness and safety of this procedure in patients with knee osteoarthritis.

Role of P4HA2 in promoting cancer progression / 国际肿瘤学杂志

P4HA2, one of the important genes encoding the α subunit of collagen prolyl 4-hydroxylase (C-P4H) , is overexpressed in a variety of tumors and promotes tumor progression, and its high expression correlates with the poor prognosis of patients. In different tumors, P4HA2 can promote the proliferation, migration and invasion of tumor cells, increase the proportion of cancer stem cell population, and promote tumor cells to escape from immune system. An in-depth understanding of the role of P4HA2 in tumor progression may provide new ideas and insights for targeting P4HA2 to prevent or reverse tumor progression.

High expression of P4HA3 in obesity: a potential therapeutic target for type 2 diabetes

The aims of the present study were to evaluate the expression of prolyl 4-hydroxylase subunit alpha 3 (P4HA3) in adipocytes and adipose tissue and to explore its effect on obesity and type 2 diabetes mellitus (T2DM). We initially demonstrated that P4HA3 was significantly upregulated in the subcutaneous adipose tissue of obesity and T2DM patients, and its functional roles in adipocyte differentiation and insulin resistance were investigated using in vitro and in vivo models. The knockdown of P4HA3 inhibited adipocyte differentiation and improved insulin resistance in 3T3-L1 cells. In C57BL/6J db/db mice fed with a high fat diet (HFD), silencing P4HA3 significantly decreased fasting blood glucose and triglycerides (TG) levels, with concomitant decrease of body weight and adipose tissue weight. Further analysis showed that P4HA3 knockdown was correlated with the augmented IRS-1/PI3K/Akt/FoxO1 signaling pathway in the adipose and hepatic tissues of obese mice, which could improve hepatic glucose homeostasis and steatosis of mice. Together, our study suggested that the dysregulation of P4HA3 may contribute to the development of obesity and T2DM.

High expression of prolyl 4-hydroxylase subunit alpha-2 in lung adenocarcinoma indicates poor prognosis

Abstract Objective To analyze the Prolyl 4-Hydroxylase subunit Alpha-2 (P4HA2) expression in Lung Adenocarcinoma (LAUD). Methods The authors assessed P4HA2 expression in the LUAD tumor ecosystem using single-cell analysis. The authors analyzed the relationship between P4HA2 expression and clinical features in LUAD and Brain Metastasis (BM) cases. The authors assessed the biological functions of P4HA2 using The Cancer Genome Atlas-LUAD dataset. Results P4HA2 was more highly expressed in fibroblasts than in epithelial cells in normal lung and lung adenocarcinoma tissues (p < 0.001). P4HA2 was more highly expressed in malignant epithelial cells than in fibroblasts in the BM tissue (p = 0.002). P4HA2 expression was significantly higher in female cases than in male cases (p = 0.049) and was related to lymph node metastasis (p = 0.019) and a higher TNM stage (p = 0.020). High P4HA2 expression indicated a poor prognosis and served as an independent prognostic risk factor in lung cancer. P4HA2 was mainly enriched in the extracellular matrix organization, NADH regeneration, and canonical glycolysis. P4HA2 expression was negatively correlated with naive B cells, T-cells, CD8, and activated natural killer cells, but positively correlated with CD4 memory-activated T cells, regulatory T-cells, resting dendritic cells, and dendritic cell activation. P4HA2 messenger RNA expression was correlated with programmed death-ligand 1 and cytotoxic T-lymphocyte-associated protein 4. Conclusion P4HA2 is highly expressed in LUAD tumor cells, especially for the BM subtype, and is a valuable prognostic indicator of LUAD. It may be involved in a biological activity of distant metastasis of LUAD tumor cells and serve as a potential treatment target.

Genetic evolution of influenza B virus in Guizhou Province during 2017 to 2021 / 中华微生物学和免疫学杂志

Objective:To analyze the molecular evolution characteristics of HA and NA genes of influenza B/Yamagata (BY) and influenza B/Victoria (BV) lineage viruses in Guizhou Province, aiming to provide reference for scientific prevention and control of influenza. Methods:The prevalence of various types of influenza viruses in Guizhou Province from 2017 to 2021 was analyzed. The nucleic acid of influenza B viruses was extracted, and then the HA and NA genes were amplified by RT-PCR. Fourteen strains were sequenced and the sequences of 83 strains were obtained from GISAID. Homologies between the 97 influenza B viruses as well as the phylogenetic characteristics and amino acid site variations were analyzed. Results:Influenza A, BY and BV lineage viruses co-circulated in Guizhou Province and BV lineage was the predominant type. The homologies of HA and NA genes were 98.7%-99.4% and 98.4%-99.6% between BY lineage viruses and the reference vaccine strain B/PHUKET/3073/2013. BV lineage viruses shared 98.3%-99.3% and 98.9%-99.6% homologies with the reference vaccine strain B/Colorado/06/2017. The BY lineage strains in Guizhou Province mainly belonged to Y3 genetic group with HA gene in two branches of Y3-H1-2 and NA gene in three branches of Y3-N1-3. Three reassortant strains were found in Y3 clade. The isolated BV lineage strains mainly belonged to V1A-2 genetic group with HA gene in four branches of V1A-2 H1-4 and NA gene in five branches of V1A-2 N1-5. Twenty reassortant strains were found in V1A-2 clade and no inter-lineage reassortants were found. Analysis of variations at key amino acid sites showed that there was no mutation at epitopes in Y3 genetic group. However, there were point mutations at four main epitopes and a shift mutation in 190 helix in V1A-2 genetic group. There was no mutation in drug resistance sites. Conclusions:Various types of influenza viruses circulated in Guizhou Province. The homology between influenza B viruses and vaccine strains was decreasing. Different branches of HA and NA genes had been evolved and various forms of mutations were detected in the sequences. Intra-lineage reassortant strains and new varieties emerged. Surveillance of influenza B viruses should be strengthened.

Hábitos alimentarios en estudiantes universitarios de la Región de Bío-Bío, Chile, 2017 / Eating habits in university students of the Bío-Bío Region, Chile, 2017

Resumen Antecedentes: las conductas poco saludables, como baja actividad física, ayuno prolongado, con sumo de alimentos de alta densidad energética y baja ingesta de frutas y verduras repercuten en la salud. Objetivo: conocer los hábitos alimentarios de estudiantes del Campus San Andrés de la Universidad Católica de la Santísima Concepción, Región del Bío-Bío, Chile. Materiales y métodos: se aplicó la encuesta dicotómica "¿Es tu alimentación saludable?" en 350 estudiantes. Resulta dos: el 75 % de los encuestados tenía una alimentación no saludable o poco saludable, con mayor prevalencia en el rango etario entre 17-20 años (78 %). Se destaca el bajo consumo de frutas (<20 %), verduras (42 %) y agua (46 %), junto con una baja prevalencia de conductas saludables/responsables, como evitar alimentos azucarados (36 %) o embutidos (38 %) y revisar los etiquetados nutriciona les (37 %). Los hombres presentaron mayor consumo de pan, carnes blancas y agua (p<0,05), mientras que las mujeres declararon en mayor porcentaje evitar embutidos, revisar los etique tados nutricionales (p<0,01), preferir alimentos azucarados y comer de forma adecuada (p<0,05). Conclusiones: los estudiantes universitarios encuestados presentan hábitos alimentarios poco salu dables, asociados principalmente a bajo consumo de frutas y verduras.

Abstract Background: Unhealthy behaviors such as low physical activity, prolonged fasting, consumption of energy-dense foods and low consumption of fruits and vegetables have repercussions on health. Objective: To understand the dietary habits of students from the San Andres campus of the Universidad Católica de la Santísima Concepción, in the Bío-Bío region of Chile. Materials and Methods: The dichotomous survey, "Is your diet healthy?" was applied to 350 students. Results: The survey revealed that 75% of the students had an unhealthy diet, with a greater prevalence in those between 17-20 years of age (78%). It also demonstrated a low consumption of fruits (<20%), vegetables (42%) and water (46%), together with a low prevalence of healthy/responsible behaviors like avoiding sugary foods (36%), avoiding processed meats (38%), and reading nutrition labels (37%). Men reported a greater consumption of bread, white meats, and water (p<0,05), while women reported the greatest percentages of avoiding processed meats, read ing nutrition labels (p<0,01), and preferred sugary foods and eating properly (p<0,05). Conclusion: The surveyed university students presented poor dietary habits, primarily associated with low consumption of fruits and vegetables.

Reseach progress of P4HA regulating solid tumor genesis and development and its mechanism / 中国肿瘤生物治疗杂志

@#[摘 要] 脯氨酰4-羟化酶α亚基(prolyl 4-hydroxylase subunit alpha，P4HA)是胶原蛋白翻译后加工的关键酶，可通过影响胶原蛋白的结构、功能及稳定性诱导肿瘤微环境重塑，从而调节肿瘤细胞极性、运动和信号转导，其与多种实体肿瘤的侵袭、转移密切相关。此外，P4HA可调控肿瘤相关基因表达、增强肿瘤细胞恶性表型表达及耐药性形成，P4HA的失调在肿瘤的生长、侵袭以及治疗抵抗中起重要作用。本文通过概述P4HA的生物学结构、功能及其在实体肿瘤发生发展中的作用机制，对P4HA与肿瘤发生、转移及耐药性关系的研究进展加以阐述。

Correlation of DAT-positive hemolytic anemia with gender, age and ABO serological indicators / 中国输血杂志

【Objective】 To explore the correlation of DAT-positive hemolytic anemia(HA) with gender, age, and ABO serological indicators (anti-A, anti-B, anti-AB, anti-D; Ac, Bc, Oc), so as to provide references for clinical diagnosis. 【Methods】 2 832 HA patients who had underwent DAT tests in the Second Affiliated Hospital of Xi′an Jiaotong University in recent years were divided into the positive-DAT patients ( n=382 )and negative-DAT patients (n=2 450 ) according to the testing results. The correlation between the positive-DAT patients and the sex, age and ABO serological indicators was analyzed using SPSS 25.0 statistical software. 【Results】 DAT-positive HA risk in women were significantly higher than men (OR=1.712, CI95%= [1.365, 2.147], P<0.01), and it differed significantly by ages; it increased significantly with the presence of positive anti-A, negative Ac (OR=1.429, CI95%= [1.150, 1.775] ; P<0.01), and positive Oc(OR=21.369, CI95%= [13.813, 33.057] ; P<0.01). Men aged 31~40 were prone to develop mixed type(OR=19.304, CI95%= [2.208, 168.751] ; P<0.01), women aged 41~50 were prone to cold antibody(OR=3.981, CI95%= [1.545, 10.259] ; P<0.01); anti-AB positive patients were prone to mixed type (OR=1.852, CI95%= [1.027, 3.339] ; P<0.05), Oc positive was prone to warm antibody(OR=4.018, CI95%= [1.386, 1.646], P<0.01). 【Conclusion】 DAT-positive HA is significantly correlated with gender and age, and is positively correlated with anti-A, Oc positive and Ac negative.

Knowledge and Attitude towards Vitamin D Deficiency in Ha’il Region: A Community-Based Study

Background: Vitamin D deficiency represents a major health problem in Saudi Arabia. Therefore, this study aimed to assess the Knowledge and attitude towards vitamin D deficiency in Ha’il Region (Northern Saudi Arabia). Methodology: A total of 500 Saudi volunteers living in the city of Ha’il and surrounding towns were included during the period from January 2020 to March 2020. Adults aged 15 years or older were randomly selected regardless of gender, occupation or marital status. Results: Out of the 500 study subjects, 214 (42.8%) did previous vitamin D testing (72/207 (34.8%) males and 142/293 (48.5%) were females). Females were significantly better for testing their vitamin D status than males (p=0.0032). Conclusion: Vitamin D deficiency is highly prevalent in Ha’il Region. Although there is a relatively higher awareness level towards vitamin D sources, vitamin D testing, as well as knowledge about vitamin D deficiency-related disorders are poor among Saudi living in Ha’il Region.

The Prevalence of Symptoms of Shift Work Disorder (SWD) among Nurses in the Northwestern Region, Saudi Arabia

Aims: This is the first study investigating the prevalence of SWD concerning depression, anxiety, insomnia, and fatigue in the Kingdom of Saudi Arabia. This study looks at the relationship between SWD Symptoms and shift work per week and work hours per week. Methodology: This study makes use of the descriptive-correlational design, and there are 250 respondents. The focus of this study is the government hospitals in Ha’il, Saudi Arabia. This study used a combination of established questionnaires which are: Hospital Anxiety and Depression Scale, Epworth Sleep Scale, and Fatigue Severity Scale. The Statistical Package for Social Sciences (SPSS) version 22 was used in the analysis of data. Frequency, percentage, and standard deviation, mean and Pearson-r was utilized. Results: The mean score for SWD symptoms are as follows: anxiety is 6.20, depression is 6.80, insomnia is 8.07, and fatigue is 35.28. On the relationship between shift work per week and SWD Pearson-r revealed scores (r=0.05, n=199, p=0.50), (r=0.11, n=199, p=0.14), (r=0.04, n=199, p=0.60), (r=0.11, n=199, p=0.14) for anxiety, depression, insomnia, and fatigue respectively. Finally for the relationship between work hours per day and SWD symptoms, Pearson-r revealed scores (r=0.25, n=199, p=0.001), (r=0.26, n=199, p=0.03), (r=0.16, n=199, p=0.001), (r=0.25, n=199, p=0.03) for anxiety, depression, insomnia, and fatigue respectively. Conclusion: This study found out that the most prevalent shift work symptom is slight dozing. Shift work per week does not influence the prevalence of shift work symptom; instead, working hours per day has a minimal influence on the shift work symptom.

Is hyaluronic acid filler still a potential risk factor for an autoimmune reaction?

@#Background: Rejuvenation of the skin with hyaluronic acid (HA) filler is considered to be one of the most favourable procedures in the field of aesthetics. Nevertheless, some adverse effects still occur though infrequently, and are associated with its use. Previous research has suggested that HA filler may stimulate antibodies. Consequently, an investigation of the immune interactions associated with use of HA filler is an important area for investigation. Objectives: The aim of this research is to investigate whether HA filler influences the initiation of an autoimmune reaction in healthy women who had received HA filler by screening for autoantibodies in the blood. Results will be compared with agematched apparently healthy control women who did not receive the filler. Methods: Serum samples were obtained from 44 females who had received HA filler and 44 females who had not as a control group. The enzyme-linked immunosorbent assay (ELISA) technique was utilised to measure serum concentrations of antiThyroglobulin (Tg), anti -thyroid peroxidase (TPO), rheumatoid factor (RF), anti-nuclear antibody (ANA) and anticentromeres. Results: The number of women who tested positive for the measured autoantibodies was not statistically significant (p=0.803) between those who had received HA filler (n=10/44, 25%) and the control group (n=11/44, 22.7%). Conclusion: Based on our result HA filler procedures do not induce an autoimmune reaction in women who received HA filler compared to controls. And consequently, HA filler procedures are relatively safe, and these results contradict the findings of other non-controlled works.

Hyaluronic acid derivative-modified nano-structured lipid carrier for cancer targeting and therapy / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

To reduce the problems of poor solubility, high in vivo dosage requirement, and weak targeting ability of paclitaxel (PTX), a hyaluronic acid-octadecylamine (HA-ODA)-modified nano-structured lipid carrier (HA-NLC) was constructed. HA-ODA conjugates were synthesized by an amide reaction between HA and ODA. The hydrophobic chain of HA-ODA can be embedded in the lipid core of the NLC to obtain HA-NLC. The HA-NLC displayed strong internalization in cluster determinant 44 (CD44) highly expressed MCF-7 cells, and endocytosis mediated by the CD44 receptor was involved. The HA-NLC had an encapsulation efficiency of PTX of 72.0%. The cytotoxicity of the PTX-loaded nanoparticle HA-NLC/PTX in MCF-7 cells was much stronger than that of the commercial preparation Taxol®. In vivo, the HA-NLC exhibited strong tumor targeting ability. The distribution of the NLCs to the liver and spleen was reduced after HA modification, while more nanoparticles were aggregated to the tumor site. Our results suggest that HA-NLC has excellent properties as a nano drug carrier and potential for in vivo targeting.