[Progress on the diagnosis and therapy about atopic diseases and primary immunodeficiency diseases].

Primary immunodeficiency diseases (PID) is a congenital disease caused by single gene germline mutation related to the immune system. PID patients have immune dysregulation, and are susceptible to infectious diseases, autoimmune diseases, autoimmune diseases, allergic diseases, and malignant tumors. The first symptom of some PID patients is atopic disease, therefore they go to the department of allergy, department of pediatrics and other relevant departments. How to identify and diagnose PID in allergic patients, to reduce diagnosis delay and prevent disease aggravation are the abilities that allergists, pediatricians, and doctors in other relevant departments need to master. This article summarizes the warning signs of PID in allergic patients and the mechanism of allergy combined with PID, and then summarizes the common types of PID in allergic patients, the evaluation, treatment and prevention in patients with PID and allergy.

[The 494th case: acute pancreatitis, acute acalculous cholecystitis and anaphylaxis].

A young male patient with abdominal pain and fever was diagnosed as acute hyper-triglyceridemicpancreatitis is clear. During the recovery of pancreatitis, the patient developed acute acalculous cholecystitis, as well as carbapenem-resistant Enterobacter infection and Cytomegaloviremia, and had anaphylaxis for several times after the use of antibiotics, which cannot be completely explained by drug allergy. This paper analyzes the possible causes of multiple diseases in the same patient in detail.

[Prevalence and risk factors of pterygium in Han and Yugur populations in Gansu Province, China].

Objective: To investigate the prevalence and risk factors of pterygium in Han and Yugur populations aged 40-79 years in Gansu Province, China. Methods: This was a cross-sectional study. A multistage cluster sampling method with urbanization level-based stratification was employed. Multiple logistic regression analysis was performed to evaluate the risk factors of pterygium. Results: A total of 4 193 people (1 840 males, 2 353 females; 3 035 Hans, 1 158 Yugurs) aged over 40 in Gansu Province were included in the study. Among them, 391 patients (9.3%) were found to have pterygium. The prevalence of pterygium adjusted for age and sex was 9.3%. The prevalence rates of Han and Yugur participants were 8.8% (267 patients) and 10.7% (124 patients), respectively, and there was no significant difference between them (χ²=3.629, P=0.057). Multivariate regression analysis showed that the risk factors of pterygium included age (OR=3.66, 95%CI: 2.26-5.92), length of residence in the countryside (OR=2.18, 95%CI: 1.41-3.38), and education level (OR=0.49, 95%CI: 0.29-0.83). In the Han group, the risk factors of pterygium were age (OR=3.84, 95%CI: 2.18-6.78) and length of rural residence (OR=2.02, 95%CI: 1.23-3.33), and a higher level of education (OR=0.36, 95%CI: 0.20-0.66) was a protective factor. Older age (OR=3.11, 95%CI: 1.13-8.59) and rural residential length ratio (OR=3.28, 95%CI: 1.09-9.88) were risk factors for pterygium in Yugur population. Conclusions: The overall prevalence of pterygium in Han and Yugur populations aged over 40 in Gansu Province, China was 9.3%, with no significant difference between the two nationalities. Older age and rural residency increased the incidence of pterygium, and a higher education level was a protective factor for pterygium.(Chin J Ophthalmol, 2020, 56:600-607).

The potential biological functions of circular RNAs during the initiation of atresia in pig follicles.

The specific expression profile and function of circular RNAs (circRNAs) in mammalian ovarian follicles, especially during the atresia process, are unclear. In this study, genome-wide deep circRNA sequencing was applied to screen circRNAs in healthy and early atretic antral follicles in pig ovaries. A total of 40,567 distinct circRNAs were identified in follicles, among which 197 circRNAs (108 upregulated and 89 downregulated) were significantly shifted during the early atresia process. Most differentially expressed circRNAs (DECs) lacked protein-coding potential. Annotation analysis of the DECs revealed 162 known host genes, or noncoding RNAs, and 10 intergenic regions. The key pathways in which these host genes are involved include the focal adhesion-PI3K-Akt-mTOR signaling pathway, vascular endothelial growth factor A (VEGFA)-vascular endothelial growth factor receptor 2 signaling pathway and transforming growth factor-beta signaling pathway. Further comparison analysis between host genes of DECs and the differentially expressed linear messenger RNA transcripts revealed the cotranscription of circRNAs and their linear mRNAs in inhibin beta units (INHBA and INHBB), glutathione S-transferase (GSTA1), and VEGFA. In addition, we predicted 196 pairs of potential circRNA-micro RNA (miRNA) interactions among 77 DECs and 101 porcine miRNAs. We have identified 16 functional miRNAs by comparing the 101 miRNAs to the functional miRNAs reported in mammal ovarian follicle atresia and granulosa cell apoptosis studies. Our study adds new knowledge to circRNA distribution profiles in pig ovarian follicles, offers a valuable reference for transcriptomic profiles in the initiation of follicular atresia, highlights warranted circRNAs for further functional investigation, and provides possible biomarkers for ovarian dysfunctions.

Upregulated exosomal miR-221/222 promotes cervical cancer via repressing methyl-CpG-binding domain protein 2.

OBJECTIVE: Methyl-CpG-binding domain protein 2, a target gene of miR-221and miR-222, plays a crucial role in a large body of human cancers. In this study, we aim to explore the mechanism by which miR-221/222 promotes cervical cancer. MATERIALS AND METHODS: We have analyzed mRNA expression of MeCP2 and MBD2 in cervical cancer tissues by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and examined the expression of miR-221/222 in C33A, HeLa, and CaSki cells by Western blot. RESULTS: We found that the expression levels of MBD2 and MeCP2 were significantly reduced in cervical cancer samples as detected by the analysis of MeCP2 in matched tumor-normal samples of patients with cervical cancer, indicating a reduction in a significant percentage of patients. At the same time, we found that miR-221 and miR-222, which targeted MBD2, were upregulated in cervical cancer samples. To further elucidate the relation between miR-221/222 and MBD2, we used a lot of cell lines such as C33A, HeLa, and CaSki. Surprisingly, we found that the expression levels of MBD2 and MeCP2 were significantly lower in HeLa and CaSki than in C33A, as detected by qRT-PCR. Western blot analysis of MeCP2 of HeLa and CaSki was significantly lower than in C33A. MiR-221/222 was significantly higher in HeLa and CaSki than in C33A by qRT-PCR. The knockdown of miR-221/222 in HeLa and CaSki resulted in the downregulation of miR-221/222 levels, which rescued the expression levels of MBD2 and MeCP2 in HeLa and CaSki. However, transfection of miR-221/222 on C33A could upregulate the expression levels of miR-221/222 and decrease the expression levels of MBD2 and MeCP2. CONCLUSIONS: Our results demonstrate that the upregulated miR-221/222 promotes cervical cancer by repressing MBD2 and MeCP2.

Targeted inhibition of ß-catenin by miR-320 and decreased MMP-13 expression in suppressing chondrocyte collagen degradation.

OBJECTIVE: Wnt/ß-catenin pathway plays a critical role in modulating embryonic development, cell growth, and differentiation. The over-expression of ß-catenin activates this pathway and up-regulates expression of matrix metalloproteinase-13 (MMP-13), and promotes matrix degradation and occurrence of osteoarthritis (OA). This study aims to explore the effect of miR-320 expression in OA chondrocyte and underlying mechanisms. PATIENTS AND METHODS: Chondrocyte tissues from OA patients and normal individuals were collected for the detection of expression levels of miR-320, ß-catenin, MMP-13, and alpha-1 chain of type II collagen (COL2A1). Dual luciferase reporter assay was performed to test targeted regulation between miR-320 and ß-catenin. IL-1ß was used to simulate in vitro cultured chondrocytes, which were transfected with miR-320 mimic and/or si-ß-catenin, followed by quantification of miR-320, ß-catenin, MMP-13, and COL2A1. RESULTS: In chondrocytes of OA patients, expression of microRNA (miR)-320 is decreased. Bioinformatics analysis revealed complementary binding sites between miR-320 and ß-catenin. Compared to control group, increasing levels of ß-catenin and MMP-13 expression with reduction of miR-320 and COL2A1 expressions were observed in OA chondrocytes. Transfection of miR-320 mimic and/or si-ß-catenin depressed expression of ß-catenin and MMP-13 inside chondrocytes, accompanied with elevation of COL2A1 expression. CONCLUSIONS: MiR-320 expression in OA chondrocyte is decreased, accompanied with up regulation of ß-catenin and MMP-13. MiR-320 can inhibit ß-catenin and MMP-13 expressions, elevates COL2A1 expression, which provides novel insights for the treatment of osteoarthritis.

[The localization diagnosis of patients with adrenocorticotropic hormone-dependent Cushing's syndrome in adolescence].

Objective: To elucidate the clinical characteristics and localization diagnosis of patients with adrenocorticotropic hormone (ACTH)-dependent Cushing's syndrome (CS) in adolescence. Methods: The clinical data, laboratory examination and localization diagnosis from 35 patients aged less than 18 years old with adolescent CS who were treated at Peking Union Medical College Hospital between January 1990 and March 2012 were analyzed. Results: There were 29 cases of Cushing's disease (CD) and 6 cases of ectopic ACTH syndrome (EAS). Compared to patients with EAS, those with CD were older at diagnosis[(15.2±2.7) vs (12.8±4.4) years], and had longer disease course[(1.9±1.5) vs (0.7±0.3) years]and higher serum potassium[(3.8±0.6) vs (2.5±0.7) mmol/L], however the plasma ACTH level[(15.4±14.9) vs (42.5±22.7) pmol/L]was lower (all P<0.05). If the cut-off of the ratio of 24-hour urine free cortisol (24 h UFC) after low-dose dexamethasone suppression test (LDDST) to before LDDST was 0.65, the sensibility to diagnose CD was 70.8%, and the specificity was 100%. If the cut-off of the 24 h UFC ratio after high-dose dexamethasone suppression test (HDDST) to before HDDST was 0.54, the sensibility to diagnose CD was 91.7%, and the specificity was 100%. If the cut-off of the plasma ACTH ratio of inferior petrosal vein[bilateral inferior petrosal sinus sampling (BIPSS)]to peripheral vein was 2, only 6 CD patients (6/8) met it. Conclusion: The study suggested that HDDST was more meaningful in the localization diagnosis of patients with ACTH-dependent CS in adolescence.

Association of single nucleotide polymorphism in melatonin receptor 1A gene with egg production traits in Yangzhou geese.

In the present study the melatonin receptor 1A gene (MTNR1A) was proposed to be a candidate gene for egg production in Yangzhou geese. A total of 210 goose blood samples were collected to investigate the association of the MTNR1A gene with the number of eggs produced. Using a direct sequencing method, a single nucleotide polymorphism (SNP; g.177G>C) was detected in the 5' regulatory region of the MTNR1A gene (Genbank ss1985399687). Two alleles (G and C) and three genotypes were identified. Association analysis results showed that the g.177G>C SNP significantly affected the level of egg production within a 34-week egg-laying period (P < 0.05). Furthermore, the geese with the GG genotype produced significantly more eggs compared to the geese with the CC genotype. Quantitative real-time PCR analysis showed that the MTNR1A gene was highly expressed in small intestine, granulosa cell and ovary compared to other examined tissues. In addition, the mRNA expression level of MTNR1A in ovary indicated that significantly higher expression levels were recorded for geese with the GG genotype compared to those with the CC genotype. Moreover, a luciferase reporter assay showed that the CC genotype had significantly lower promoter activity than did GG. These results suggest that the identified SNP in the MTNR1A gene may influence the number of eggs produced and mRNA expression levels in Yangzhou geese and could be considered as a useful molecular marker in goose selection and improvement, especially for egg production.

Effect of progranulin (PGRN) on the proliferation and senescence of cervical cancer cells.

We investigated the effect of progranulin (PGRN) expression on the proliferation and senescence of cervical cancer cells. PGRN small interfering RNA (siRNA) was introduced into the SiHa and HeLa cell lines of human cervical carcinoma using liposome-mediated transfection. The expression levels of PGRN in each cell line after transfection of PGRN siRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR). Senescence in the cell lines was detected using the ß-galactosidase-staining test, and proliferation was detected by clone formation. The RT-PCR assay showed that the expression of PGRN in all of the cell lines transfected with PGRN siRNA markedly decreased. In the clone-forming test, compared with the control group, the colony-forming ability in all cell lines decreased significantly after transfection with PGRN siRNA. The ß-galactosidase-staining experiments showed that the phenomenon of cell aging in the PGRN interference group was more obvious than in the control group. After the cervical cancer cells had been transfected with PGRN siRNA, cell senescence was accelerated and clone-forming ability was markedly reduced. This suggests that PGRN can promote the proliferation of the cervical cancer cell line; proliferation of cervical cancer cells is achieved by inhibiting their senescence.

Molecular cloning of the goose ACSL3 and ACSL5 coding domain sequences and their expression characteristics during goose fatty liver development.

It has been demonstrated that ACSL3 and ACSL5 play important roles in fat metabolism. To investigate the primary functions of ACSL3 and ACSL5 and to evaluate their expression levels during goose fatty liver development, we cloned the ACSL3 and ACSL5 coding domain sequences (CDSs) of geese using RT-PCR and analyzed their expression characteristics under different conditions using qRT-PCR. The results showed that the goose ACSL3 (JX511975) and ACSL5 (JX511976) sequences have high similarities with the chicken sequences both at the nucleotide and amino acid levels. Both ACSL3 and ACSL5 have high expression levels in goose liver. The expression levels of ACSL3 and ACSL5 in goose liver and hepatocytes can be changed by overfeeding geese and by treatment with unsaturated fatty acids, respectively. Together, these results indicate that ACSL3 and ACSL5 play important roles during fatty liver development. The different expression characteristics of goose ACSL3 and ACSL5 suggest that these two genes may be responsible for specific functions.

miR-34a targets the inhibin beta B gene, promoting granulosa cell apoptosis in the porcine ovary.

During ovarian follicular growth and development, only a few follicles actually ovulate. Recently, it was found that follicular atresia is triggered by granulosa cell apoptosis, but the molecular mechanism of follicular atresia was not understood. Using flow cytometry, we found that miR-34a promotes granulosa cell apoptosis in pig ovarian follicles. In addition, inhibin beta B was found to be a miR-34a target gene, based on luciferase reporter assays, quantitative RT-PCR and Western blotting. Taken together, our data indicate that miR-34a plays an important role in granulosa cell apoptosis by targeting the INHBB gene in the porcine ovary.

Effect of cholesterol on lipogenesis and VLDL-TG assembly and secretion in goose primary hepatocytes.

To investigate how cholesterol induces hepatocytic steatosis, we investigated the effect of cholesterol on hepatic lipogenesis and the assembly and secretion of very-low-density lipoprotein-triglycerides (VLDL-TGs) in goose primary hepatocytes. We found that cholesterol at 20 µg/ml increased the concentrations of extracellular VLDL, intracellular cholesterol, and intracellular TGs, while cholesterol at 30 µg/ml had a reduced effect (p < 0.05). Additionally, cholesterol at 20 µg/ml, but not at 10 or 30 µg/ml, increased the extracellular TG concentration. Cholesterol increased the fatty acid synthase (FAS) enzyme activity in a dose-dependent manner. Incubation with cholesterol increased the mRNA level of genes involved in lipogenesis, including sterol regulatory element-binding proteins (SREBPs), FAS, acetyl-CoA carboxylase-α (ACCα), and liver X receptors. The mRNA level of the acyl-CoA: diacylglycerol acyltransferase 1 (DGAT1) gene changed in response to cholesterol treatment in a dose-dependent manner. Similar to the regulation of extracellular VLDL and intracellular TG accumulation, the mRNA levels of the microsomal triglyceride transfer protein, forkhead box O1, and DGAT2 increased with treatment with 10 or 20 µg/ml of cholesterol, but decreased with treatment with 30 µg/ml of cholesterol (p < 0.05). Cholesterol had no evident effect on the mRNA level of the apolipoprotein B gene. Incubation with cholesterol at 20 and 30 µg/ml increased the nuclear SREBP-1 protein level (p < 0.05) and the binding affinity of the nuclear SREBP-1 to ACCα SRE probes. In conclusion, cholesterol not only activates the transcription of genes involved in fatty acid synthesis and TG accumulation, but also activates the transcription of genes involved in the assembly and secretion of VLDL-TG in goose primary hepatocytes.

Efficient CdSe quantum dot-sensitized solar cells prepared by a postsynthesis assembly approach.

A postsynthesis assembly approach, an ex situ ligand exchange route, was developed for fast (within 2 h) and high loading (34% coverage) deposition of CdSe QDs on TiO(2) films. With the combination of high-quality QD sensitizers and the effective deposition technique, a record photovoltaic performance with an efficiency of 5.4% was observed for the resulting cell device.

Comparison and characteristics of the formation of different adipose tissues in ducks during early growth.

The objective of the current research was to investigate the pattern of subcutaneous adipose tissue growth during Peking duck (Anas platyrhynchos) early development and to determine the reasons for regional differences. The morphological characteristics in 5 regions of subcutaneous tissue, including the neck area (NSF), chest area (CSF), lower abdomen area (ASF), back area (BSF), and leg area (LSF), were analyzed by comparing the morphology of the sections, adipocyte volume and number, and lipid content from wk 1 to 8. Moreover, the mRNA expression of several molecular marker genes, including 47-kDa tail interacting protein (TIP47), adipose differentiation-related protein (ADRP), and perilipin, were detected from wk 1 to 8 using quantitative real-time PCR. Our results revealed that the average cell number declined greatly as fattening proceeded (except in the NSF) and changed very little after wk 4 in all 5 regions. In contrast, the average cell volume and triglyceride content per cell increased gradually during early duck growth. The BSF and LSF lipid content had a different pattern of change than the other regions. The NSF, CSF, and ASF regions had the highest lipid content values at all stages, the BSF was intermediate, and the LSF was the lowest at all weeks except wk 3. The highest TIP47 expression level was found in the NSF from wk 1 to 2 and BSF at wk 1. The relative expression level of TIP47 was higher in the CSF than in the ASF and BSF at wk 4, and was higher in the NSF than in the ASF at wk 6. The highest levels of ADRP and perilipin were detected in the LSF. These results suggest that a combination of adipocyte hyperplasia and hypertrophy is mainly responsible for the development of duck adipose tissue before wk 4, after which adipose expansion is accomplished by adipocyte hypertrophy only. Adipocyte hyperplastic and hypertrophic capacity, fat storage capacity, and metabolic activity may be partial explanations for the regional differences during duck growth.

Effect of liver X receptor activation on the very low density lipoprotein secretion and messenger ribonucleic acid level of related genes in goose primary hepatocytes.

In this study, we investigated the role of liver X receptor (LXR) activation in hepatic assembly and in the secretion of very low density lipoprotein-triglycerides in goose primary hepatocytes. Goose primary hepatocytes were isolated and treated with the LXR agonist T0901317. Total triglyceride accumulation, intracellular and extracellular triglyceride concentrations, extracellular very low density lipoprotein concentration, and gene expression levels of LXRα, microsomal triglyceride transfer protein, acyl coenzyme A:diacylglycerol acyltransferase (DGAT) 1, and DGAT2 were measured in primary hepatocytes. We found a dose-dependent upregulation of total and intracellular TG accumulation when using 0, 0.01, 0.1, 1, and 10 µM T0901317, but the extracellular triglyceride and very low density lipoprotein concentrations were dose dependent only when the T0901317 concentration was below 1 µM; as compared with 1 µM T0901317, 10 µM T0901317 had an inhibiting effect (P < 0.05). The mRNA levels of all the detected genes increased in the presence of T0901317. The change in LXRα and DGAT1 was dose dependent, and the mRNA levels of microsomal triglyceride transfer protein and DGAT2 increased with a T0901317 concentration up to 1 µM, but decreased when treated with 10 µM T0901317 (P < 0.05). In conclusion, the secretion of very low density lipoprotein plays a role in pharmacologically activating the LXR-induced development of hepatocellular steatosis in geese.

Quantification of urine 17-ketosteroid sulfates and glucuronides by high-performance liquid chromatography-ion trap mass spectroscopy.

The measurement of androgen steroids has been utilized as a clinical indicator of adrenal function, androgen abuse, and as a prediction of general health or biological aging. An improved high-performance liquid chromatography-ion trap mass spectroscopic method with sonic spray ionization (SSI) technology for the quantification of individual urinary 17-ketosteroid sulfates and glucuronides was developed and validated. Sample preparation was simplified using a C18 cartridge followed by direct injection onto a reversed-phase HPLC column. Individual 17-ketosteroid from 63 urinary specimens collected in a 24-h period was measured. 17-Ketosteroid conjugates, total 17-KS-S and the ratio of total 17-KS-S to creatinine referred to herein as the Anabolic/Catabolic Index (ACI) showed statistically significant negative correlations with age.

Structural asymmetry in the CTP-liganded form of aspartate carbamoyltransferase from Escherichia coli.

The protein and solvent structure of the CTP-liganded form of aspartate carbamoyltransferase from Escherichia coli yields an R-factor of 0.155 for data to a resolution of 2.6 A. The model has 7353 protein atoms, 945 sites for solvent, and two molecules of CTP. A total of 25 of the 912 residues of the model exist in more than one conformation. The root-mean-square deviation of bond lengths and angles from their ideal values is 0.013 A and 2.1 degrees, respectively. The model reported here reflects a correction in the trace of the regulatory chain. One molecule of CTP binds to each of the two regulatory chains of the asymmetric unit of the crystal. The interactions between the pyrimidine of each CTP molecule and the protein are similar. The 4-amino group of CTP binds to the carbonyl groups of residues 89 (tyrosine) and 12 (isoleucine) of the regulatory chain. The nitrogen of position 3 of the pyrimidine binds to the amide group of residue 12; the 2-keto group binds to lysine 60. The 2'-OH group of the ribose forms hydrogen bonds with lysine 60 and the carbonyl group of residue 9 (valine). The binding of the phosphate groups of CTP to the regulatory chain probably reflects an incomplete association of CTP with the enzyme at pH 5.8. A lattice contact influences the interaction between the triphosphate group of one CTP molecule and the protein. For the other CTP molecule, only lysine 94 binds to the phosphate groups of CTP. Of the two regulatory and two catalytic chains of the asymmetric unit of the crystal, there are only two significant violations of non-crystallographic symmetry. The active site in the vicinity of arginine 54 of one catalytic chain is larger than the active site of its non-crystallographic mate. The "expanded" cavity accommodates four solvent molecules in the vicinity of arginine 54 as opposed to two molecules of water for the "contracted" cavity. Furthermore, arginine 54 in the "expanded" pocket adopts two conformations, either hydrogen-bonding to glutamate 86 or to the phenolic oxygen atom of tyrosine 98; residues 86 and 98 are in a catalytic chain related by 3-fold symmetry to the catalytic chain of arginine 54. In the "contracted" pocket, arginine 54 binds only to glutamate 86.(ABSTRACT TRUNCATED AT 400 WORDS)