[Analysis of etiology and complications in children with stage 5 chronic kidney disease].

Objective: To investigate the etiology, complications, and prognostic factors of stage 5 chronic kidney disease (CKD5) in children. Methods: A case series study was conducted to retrospectively analyze the general situation, clinical manifestations, laboratory tests, genetic testing, and follow-up data (until October 2022) of 174 children with CKD5 who were diagnosed and hospitalized at the Children's Hospital of Chongqing Medical University from April 2012 to April 2021. The characteristics of complications in the children were compared based on age, gender, and etiology. Based on the presence or absence of left ventricular hypertrophy (LVH), patients were divided into LVH group and non LVH group for analyzing the influencing factors of cardiovascular disease. Patients were also divided into death group and survival group, peritoneal dialysis group and hemodialysis group based on the follow-up data for analyzing the prognostic factors. The chi-square test, independent sample t-test, Fisher exact probability test, Mann-Whitney U test and Kruskal Wallis test were used to analyze data among different groups. Multivariate Logistic regression analysis was used to identify the prognostic factors. Results: A total of 174 children with CKD5 were enrolled in the study (96 boys and 78 girls), aged 11.2 (8.2, 13.0) years. Congenital kidney and urinary tract malformations (CAKUT) were the most common causes of the CKD5 (84 cases, 48.3%), followed by glomerular diseases (83 cases, 47.7%), and among which 28 cases (16.1%) were hereditary glomerular diseases. The common complications of CKD5 included anemia (98.2%, 165/168), mineral and bone disorder in chronic kidney disease (CKD-MBD) (97.7%, 170/174), lipid metabolism disorders (87.5%, 63/72), hypertension (81.4%, 127/156) and LVH (57.6%,57/99). The incidences of hypertension in primary glomerular disease were higher than that in CAKUT(93.8%(30/32) vs.73.7%(56/76),χ2=5.59,P<0.05). The incidences of hypertension in secondary glomerular disease were higher than that in CAKUT and that in hereditary kidney disease (100.0%(20/20) vs. 73.7%(56/76), 68.2%(15/22), both P<0.05). The incidence of hypocalcemia in CAKUT, primary glomerular disease, and hereditary kidney disease was higher than that in secondary glomerular disease (82.1%(69/84), 88.2%(30/34), 89.3%(25/28) vs. 47.6%(10/21), χ2=10.21, 10.75, 10.80, all P=0.001); the incidence of secondary hyperparathyroidism in women was higher than that in men (80.0%(64/80) vs. 95.0%(57/60), χ2=6.58, P=0.010). The incidence of LVH in children aged 6-<12 was higher than that in children aged 12-18 (73.5%(25/34) vs. 43.1%(22/51), χ2=7.62, P=0.006). Among 113 follow-up children, the mortality rate was 39.8% (45/113). Compared to the survival group, the children in the death group had lower hemoglobin, higher blood pressure, lower albumin, lower alkaline phosphatase and higher left ventricular mass index ((67±19) vs. (75±20) g/L, 142 (126, 154) vs. 128(113, 145) mmHg(1 mmHg=0.133 kPa), (91±21) vs. (82±22) mmHg, 32 (26, 41) vs. 40 (31, 43) g/L, 151 (82, 214) vs. 215 (129, 37) U/L, 48 (38, 66) vs. 38(32, 50) g/m2.7,t=2.03, Z=2.89, t=2.70, Z=2.49, 2.79, 2.29,all P<0.05), but no independent risk factors were identified (all P>0.05). The peritoneal dialysis group had better alleviation for anemia, low calcium, and high phosphorus than the hemodialysis group ((87±22) vs. (72±16) g/L, (1.9±0.5) vs. (1.7±0.4) mmol/L, (2.2±0.7) vs. (2.8±0.9) mmol/L, t=2.92, 2.29, 2.82, all P<0.05), and the survival rate of the peritoneal dialysis group was significantly higher than that of the hemodialysis group (77.8% (28/36) vs. 48.4% (30/62), χ2=8.14, P=0.004). Conclusions: CAKUT is the most common etiology in children with CKD 5, and anemia is the most common complication. The incidence of complications in children with CKD 5 varies with age, gender and etiology. Anemia, hypertension, hypoalbuminemia, reduced alkaline phosphatase and elevated LVMI may be the prognostic factors in children with CKD5. Peritoneal dialysis may be more beneficial for improving the long-term survival rate.

[Expression profile of circular RNA in inflammatory response in human bronchial epithelial cells induced by carbon black nanoparticles].

Objective: To explore the toxic effect of carbon black nanoparticles on human bronchial epithelial cells, and identify the differentially expressed circular RNA based on the full transcriptome high-throughput sequencing, so as to provide evidence for the development of biomarkers exposed to carbon black nanoparticles and their application on epigenetic toxicology. Methods: In June 2020, 16 HBE cells were treated with carbon black nanoparticles at concentrations of 20, 40 and 80 µg/ml, and 16 HBE cells without any intervention were used as the control group. The cytotoxicity of carbon black nanoparticles was detected by CCK8 and LDH experiments. Real-time quantitative fluorescent PCR (qRT-PCR) and ELISA were used to detect the changes of interleukin-6 (IL-6) and interleukin-8 (IL-6, IL-8) mRNA and protein levels of carbon black nanoparticles with concentration gradient after 72 h exposure. Western blot analysis was conducted to detect the expression levels of toll-like receptor 4 (TLR4), phosphorylated nuclear factor-κB (P-NF-κB), apoptosis-related speckled protein (ASC) and Caspase-1 associated with nuclear factor-κB. According to high-throughput sequencing results, differentially expressed Circrnas were screened and identified by qRT-PCR, and those with stable differentially expressed circrnas and the strongest association with the NF-κB pathway were selected for ring performance identification. Results: After being exposed to carbon black nanoparticles for 72 h, the activity of 16HBE cells decreased significantly (P<0.05), and the release of lactate dehydrogenase increased significantly (P<0.05). Compared with control group, mRNA expression levels of IL-6 and IL-8, protein levels of IL-6 and IL-8 were increased, and protein levels of TLR4, p-NF-κB, ASC and Caspase-1 were significantly up-regulated in 16 HBE cells of different concentrations, with statistical significance (P<0.05). Compared with the control group, a total of 492 differentially expressed circular Rnas (|log2 FC|>1) were detected. Among the 5 differentially expressed (P<0.05) circular Rnas, circ_002642 was selected as the object of subsequent research on circular Rnas, affter 72 hours of exposure to 80 µg/ml CBNPs, 16HBE cells showed signlficantly higher expression of circ_002642 (P<0.05) . Conclusion: Carbon black nanoparticles can induce differentially expressed circular RNAs associated with inflammatory response in human bronchial epithelial cells.

Vitamin D Binding Protein and Vitamin D Levels in Multi-Ethnic Population.

INTRODUCTION: Low levels of 25-hydroxyvitamin D (25(OH)D) has been associated with many negative health outcomes including falls and fractures. 25(OH)D is largely bound to vitamin D binding protein (VDBP). There is increasing evidence that free or bioavailable 25(OH)D may be a better measure of vitamin D deficiency. OBJECTIVE: To determine the prevalence of 25(OH)D deficiency and VDBP levels in multi-ethnic population, and its impact on muscle strength. DESIGN AND METHODS: Cross-sectional study of older adults in Western region of Singapore. 295 participants from three ethnic groups were selected from the Healthy Older People Everyday (HOPE) cohort for measurements of total 25(OH)D and VDBP levels. Total 25(OH)D, VDBP, frailty status, Timed-Up-and-Go (TUG) and grip strength (GS) were assessed. Albumin, free and bioavailable 25(OH)D were only available for 256 participants. RESULTS: 53% of Malay and 55% of Indians were deficient in 25(OH)D compared with 18.2% of ethnic Chinese participants. Chinese also had higher total 25(OH)D concentrations with a mean of 29.1 ug/l, (p = <0.001). Chinese had the lowest level of VDBP (169.6ug/ml) followed by Malay (188.8 ug/ml) and Indian having the highest (220.1 ug/ml). Calculated bioavailable and free 25(OH)D levels were significantly higher in Chinese, followed by Malays and Indians, which also correlated with better grip strength measures amongst the Chinese. CONCLUSION: The Malays and Indians had overall lower free, bioavailable and total 25(OH)D compared with ethnic Chinese. Chinese ethnic group also had the lowest VDBP and better overall grip strength.

Three-dimensional digital visible heart model and myocardial pathological characteristics of fetal single ventricle connected with aortic coarctation.

This study aimed to provide data for imaging diagnosis and clinical surgical plans by reconstructing a three-dimensional (3-D) digital visible heart model of single ventricle (SV) connection with aortic coarctation (CoA) and characterizing the myocardial and vascular wall pathological characteristics. Fifteen miscarried fetus cadavers with SV and CoA were selected. Fourteen cardiac specimens were systematically reviewed for segmental anatomy and conventional histological examinations. One fetus cadaver was used to obtain the structural dataset of the fetal body and to reconstruct a 3-D digital visible heart model. Specimen pathological dissection indicated hypertrophic myocardium SV, significant aortic wall thickening, and localized coarctation area elevation. Ten cases of SV with left ventricular morphology displayed a large muscle ridge and solitus normally aligned great arteries. Five cases of SV with right ventricular morphology had coarse, parallel trabeculations and received a common atrioventricular valve. The reconstructed 3-D heart and the main internal structures were realistic, which were beneficial for clinical and image teaching of fetal heart development. The change of characteristics of the myocardium and great vascular wall was obvious and may be the critical cause leading to progressive dysfunction in the postnatal heart.

Inflammasome gene expression alterations in Staphylococcus aureus biofilm-associated chronic rhinosinusitis.

BACKGROUND: The role of inflammasomes in chronic inflammation has been the subject of intense research in recent years. Chronic rhinosinusitis (CRS), a persistent inflammatory disease, continues to be investigated hoping that a clearer pathophysiologic description will guide discovery of future treatment modalities. This study investigates the role of inflammasome complexes in CRS patients with Staphylococcus aureus biofilm infection, a key culprit associated with disease severity and recalcitrance. METHODOLOGY: Sinonasal tissue samples were collected from CRS patients with (P+) and without (P-) polyps and controls. S. aureus biofilm status was obtained using fluorescence in situ hybridization and classified as biofilm positive (B+) or negative (B-). RNA was analysed using a Human Inflammasome PCR array, profiling the expression of 84 genes involved in inflammasome function. RESULTS: Sixteen samples were obtained: 5 B+P+, 5 B-P- and 6 controls. Comparing B+P+ vs. controls showed the greatest number of differentially expressed genes. In particular, Absent in Melanoma 2 (AIM2) was consistently and significantly up-regulated in the B+P+ vs. B-P- and controls. In contrast, when comparing the B-P- vs. controls, no genes showed significant changes. CONCLUSION: Our results indicate the involvement of inflammasome complexes and their signalling pathways in CRS patients with polyps and S. aureus biofilms. In particular, AIM2, activated by intracellular double-stranded DNA, is up-regulated in this group, implying that S. aureus may play a role in intracellular triggering of the inflammasome response. Studies with further patient stratification and assessing corresponding protein expression are needed to further characterize the role of inflammasomes in CRS.

Lysozyme expression is increased in the sinus mucosa of patients with chronic rhinosinusitis.

BACKGROUND: The presence of fungi and bacteria in the paranasal sinuses may contribute to ongoing inflammation. Lysozyme is an innate immune peptide with bactericidal and fungicidal activity. The expression of lysozyme in chronic rhinosinusitis (CRS) is poorly understood and deficiencies in lysozyme expression may contribute to the ongoing inflammation in CRS patients. OBJECTIVE: Determine lysozyme expression in sinus mucosa of normal and CRS patients with (CRSwNP) and without (CRSsNP) nasal polyps. METHODOLOGY: Sinus mucosa specimens (n = 82) were processed for standard histology, immunohistochemical localisation of lysozyme, immunofluorescent localisation of fungi, and qPCR analysis of lysozyme expression. RESULTS: CRS specimens displayed high-levels of lysozyme immunoreactivity in many of the abundant serous cells. Moderate levels were detected in some epithelial cells and inflammatory cells. Low levels were detected in some subepithelial glands of control specimens. No difference in immunoreactivity was detected between CRSwNP and CRSsNP specimens. Fungal elements were not visualised in any sinus specimen. qPCR analysis demonstrated variable lysozyme expression between individuals. CONCLUSIONS: Lysozyme protein expression is increased in patients with CRS, suggesting a defect in lysozyme expression is not responsible for the microbial colonisation often associated with CRS. The functional activity of lysozyme in CRS patients needs to be further investigated.

Diversity of immunoglobulin E-encoding transcripts in sinus mucosa of subjects diagnosed with non-allergic fungal eosinophilic sinusitis.

BACKGROUND: The role of allergy in the aetiopathogenesis of chronic rhinosinusitis (CRS) remains controversial. For example, in some cases with sinus fungal infections allergy can be demonstrated by standard tests. In other cases, such signs can be absent despite elevated levels of IgE-positive cells in sinus tissue and the presence of IgE and eosinophils in the sinus mucous. OBJECTIVE: To define the nature of molecular diversity in antibodies of the IgE isotype at the site of local inflammation in subjects diagnosed with non-allergic fungal eosinophilic sinusitis (NAFES). METHODS: The local occurrence and sequence characteristics of IgE-encoding transcripts in NAFES patients were investigated and compared with sequences found in subjects diagnosed with CRS featuring systemic allergy. These sequences have also been compared with other reported IgE-encoding transcriptomes. Results IGHV genes derived from major subgroups 1, 3, 4 and 5 and a diverse set of IGHD and IGHJ genes were shown to create the IgE repertoire in patients diagnosed with NAFES and CRS. The average lengths of the third hypervariable loop in these populations were 15.8 and 14.6 residues. The sequences showed evidence of extensive somatic hypermutation (mutation frequency: NAFES, 6.4 ± 3.2%; CRS, 7.0 ± 4.4%) with substitutions targeted to complementarity-determining regions. These sequence collections thus show extensive similarities to those found in other polyclonal Ig repertoires including those encoding IgE. CONCLUSION AND CLINICAL RELEVANCE: We conclude that sinus IgE-encoding transcripts in subjects diagnosed with NAFES show evidence of conventional IgE responses and we suggest that allergens with characteristics of classical antigens should be investigated for a role in the local response occurring in NAFES. This investigation illustrates that assessment of local immunity might be an important diagnostic tool in conditions like NAFES with no systemic signs of allergy to identify or rule out an allergic component of the patient's disease.

Reflux changes in adenoidal hyperplasia: a controlled prospective study to investigate its aetiology.

OBJECTIVES: To compare pepsin, carbonic anhydrase III (CAIII), cyclooxygenase-2 (COX-2) and mucin 5AC (MUC5AC) expression in children with adenoid hypertrophy and normal controls. DESIGN: A non-randomised, controlled prospective study. SETTING: Two paediatric hospitals in Adelaide, South Australia. PARTICIPANTS: Children aged 2-10 years, 21 undergoing adenoidectomy and 12 controls undergoing routine dental surgery. MAIN OUTCOME MEASURES: We measured expression of pepsin, CAIII, COX-2 and MUC5AC levels by real-time RT-PCR, immunohistochemistry, and Western blot to determine any difference between children with hyperplastic adenoids and controls. RESULTS: Pepsin was not detected in any study or control adenoid by immunohistochemistry or Western blot. Real-time RT-PCR analysis showed a statistically significant difference between groups with respect to COX-2 (P = 0.027) and MUC5AC (P = 0.02) but no difference in CAIII expression (P = 0.414). A significant correlation was also found between COX-2 and MUC5AC expression (Kendall Tau = 0.4, P = 0.005). CONCLUSION: Our results suggest that the biochemical changes seen in adenoid hypertrophy are different to those seen in reflux-affected tissues. The decreased COX-2 and MUC5AC expression may be due to squamous metaplasia and other inflammatory changes associated with adenoid hypertrophy. Our findings infer there is little evidence of reflux being a major contributory factor in the pathophysiology of adenoidal hypertrophy.

Thin sectional anatomy, three-dimensional reconstruction and visualization of the heart from the Chinese Visible Human.

The research aimed to provide sectional anatomic and three-dimensional (3D) virtual anatomic bases for imaging diagnosis and surgical operation by the use of data from the heart of the first Chinese digitized Visible Human. Data from the series of thin sections of the heart were analyzed and input into an SGI workstation, and 3D reconstruction and virtualization of the heart were performed. Each image of sectional anatomy was clear and the 3D structures of the heart were reconstructed in their entirety. All reconstructed structures can be displayed by multiple structural and color modes, individually or jointly, and can be rotated continuously in any plane. The model of the virtual heart clearly showed fine structures of the heart in random orientation. The dataset of the sectional anatomy provides a fine and integrated morphologic base for imaging diagnosis. The 3D reconstructed images clearly show the internal and entire structures of the heart.

Runoff quality and pollution loadings from a tropical urban catchment.

Runoff quality draining from 17.14 km2 urban catchment in Johor Bahru, Malaysia, was analysed. The land-use consists of residential (30.3%), agricultural (27.3%), open space (27.9%), industrial (8.1%) and commercial (6.4%) areas. Three storm events were sampled in detail. These storms produced stormflow between 0.84 mm and 27.82 mm, and peakflow from 2.19 m3/s to 42.36 m3/s. Water quality showed marked variation during storms especially for TSS, BOD and COD with maximum concentrations of 778 mg/l, 135 mg/l and 358 mg/l, respectively. Concentrations of TOC, DOC, NH3-N, Fe and level of colour were also high. In general, the river quality is badly polluted and falls in Class V based on the Malaysian Interim National Water Quality Standards. Event Mean Concentrations (EMC) for various parameters varied considerably between storms. The largest storm produced higher EMC for TSS, NO3-N and SS whereas the smaller storms tend to register higher EMC for BOD, COD, NH3-N, TOC, Ca, K, Mg, Fe and Zn. Such variations could be explained in terms of pollutant availability and the effects of flushing and dilution. Based on a three-month average recurrence interval (ARI) of rainfall, the estimated event loadings (ton/ha) of TSS, BOD, COD, TOC, NH3-N and NO3-N were 0.055, 0.016, 0.012, 0.039, 0.010, 0.0007 and 0.0002, respectively. Heavy metals present in trace quantities. Storms with 3 months ARI could capture about 70% of the total annual loads of major pollutants.

The pre-styloid compartment of the parapharyngeal space: a three-dimensional digitized model based on the Chinese Visible Human.

To build a digitized visible model of the parapharyngeal space of the Chinese Visible Human and to provide a sectional anatomic basis for radiological and clinical diagnosis of the parapharyngeal space, sectional anatomy data of the parapharyngeal space were selected from the Chinese Visible Human male and female to compare with MR imaging findings in the axial planes. From these data the parapharyngeal space and surrounding structures were segmented. They were then reconstructed in three dimensions on PC. In the axial planes of the sectional anatomy and MR imaging, the shape, content and relations of the parapharyngeal space were clearly displayed and the dominant plane for showing the parapharyngeal space was elicited. The three-dimensional reconstructed images displayed perfectly the anatomic relationships of the parapharyngeal space, parotid, muscles, mandible and vessels. All reconstructed structures can be displayed singly, in groups or as a whole; any diameter or angle of the reconstructed structures can be easily measured. The Chinese Visible Human male and female data set can provide complete and accurate data. The digitized model of the parapharyngeal space and its surroundings offers unique insights into the complex anatomy of the area, providing morphologic data for imaging diagnosis and surgery of the parapharyngeal space.

Visualization of the temporal bone of the Chinese Visible Human.

Temporal bone anatomy is very difficult to understand. After dataset acquisition of the first Chinese Visible Human, we processed the two-dimensional images to build a digitized visible model of the temporal bone and explore the role of virtual endoscopy in the inner ear. On a SGI workstation three-dimensional computer reconstructions of the ear were generated from the Chinese visible human dataset, viewing the middle and inner ear imitating the traditional otoscopy. The three-dimensional data of the temporal bone were then converted to STL format, and the temporal bone replica were fabricated with rapid prototyping by laminated object manufacturing. The virtual model of the ear was successfully completed, and the virtual endoscopy improved three-dimensional visualization of the middle and inner ear. Physical replica of the temporal bone were built with paper; the accuracy was +/-0.2 mm. The reconstructed model and the replica of the temporal bone can be used to make preoperative plans in the complicated otoneurosurgical operations, allowing various surgical exercises to be carried out on the three-dimensional stereophysical model. The virtual endoscopy stands as a promising new visualization technique for elucidation of the middle and inner ear and reveals a tremendous potential in both clinical and educational settings, providing morphological data for the image diagnosis and otoneurosurgery.

Computerized 3D-reconstructions of the ligaments of the lateral aspect of ankle and subtalar joints.

3D-reconstruction images of the structures of lateral aspect of the ankle and subtalar joints were produced using plastination to make equidistant serial sections of 1.2 mm in thickness. A SGI workstation was employed to reconstruct the structures of the ligaments of the lateral aspect of ankle and subtalar joints in three dimensions. Reconstructed structures were displayed singly, in groups or as a whole, and these were rotated continuously at different velocities in 3D space. Different diameters and angles of the reconstructed structures could be measured easily. Improved results could be achieved with the use of a special sectional anatomical technique, i.e. contours + marching cubes algorithm.

[Clinical and experimental study on yi-gan-ning granule in treating chronic hepatitis B].

This paper reports that 320 patients with chronic hepatitis B (CHB) were treated with Yi-ganning Granule (YGNG) and the pharmacodynamics of YGNG in the animal study. As control, another 70 patients with CHB receiving oleanolic acid granule (OAG) were compared to 68 patients in YGNG group. YGNG is consisted of Astragalus membranaceus , Artemisia capillaris, Codonopsis pilosula, et al. Each patient has taken YGNG or OAG for 3 months. The result showed YGNG was effective on recovering the liver function and OAG had similar effect. The sero-negative conversion rates of HBsAg, HBeAg, HBcAb and positive conversion rate of HBeAb in the YGNG group were 33.1%, 40.5%, 10.5% and 15.5% respectively, which were much better than that in OAG group (P < 0.05-0.001). The result of 6 months follow up showed that 60 of 62 patients receiving YGNG were in stabilized state. The result in the animal study demonstrated that YGNG had significant protection from the liver damage caused by CCl4. YGNG could decrease serum ALT level and protect the liver function of carbohydrate, fat, protein metabolism and detoxication. YGNG could induce interferon in vivo and play an important role in seroconversion of negative DHBV-DNA and improvement of pathological morphology in viral hepatitis B.

Development of a highly sensitive assay, based on the polymerase chain reaction, for rare B-lymphocyte clones in a polyclonal population.

A method has been developed to use the polymerase chain reaction to amplify and sequence the chain determining region 3 (CDR 3) of the human immunoglobulin heavy-chain gene, and to use the sequence as a marker for rare neoplastic B lymphocytes. Consensus primers for the Variable and Joining regions of the gene were constructed and shown to enable efficient amplification, directed cloning, and sequencing of CDR 3. Using leukaemic cell line PFMC as a test system, CDR 3 was sequenced, specific primers synthesized, and PFMC DNA was detected down to a dilution of 1:1300 in DNA from normal lymphocytes. This strategy should be useful for monitoring therapy and detecting early disease relapse in B lymphoproliferative disease.

[Effects of oleanolic acid on immune system and type I allergic reaction].

Oleanolic acid (OLA)50, 100 mg/kg sc antagonised the inhibitory effect of cortisone on the weights of spleen and thymus, decreased the clearance rate of charcoal particles in mice. The production of serum specific antibody hemolysin was not clearly affected by OLA. The administration of OLA 100 mg/kg enhanced the development of serum antibody immunoglobulin G in mice and showed marked inhibition on the hemolytic activity of the total complement by classical pathway in guinea pigs. In addition, the homologous passive cutaneous anaphylaxis in mice or rats and the degranulation of mast cells of calvarial periosteum in rats were significantly prevented by OLA 50, 100 mg/kg. The increased capillary permeability in the rat skin caused by intradermic injection of histamine was also reduced by OLA 100 mg/kg. These results indicate that OLA inhibited the type I allergic reaction.