[Short-term efficacy of posterior ankle arthroscopy and tendoscope on ankle-derived flexor hallucis longus ganglion].

Objective: To investigate the short-term clinical efficacy of posterior ankle arthroscopy combined with tendoscope on ankle-origin flexor hallucis longus tendon ganglion. Methods: A follow-up study. Clinical data of 10 patients with hallux ganglion underwent posterior ankle arthroscopy combined with flexor hallucis longus tendoscope in the Tianjin First Central Hospital from January to June 2021 were analyzed retrospectively. There were 5 males and 5 females with a mean age of 42.7 (22-54) years. Disease distribution: 6 patients were on the right side and 4 cases were on the left side. The mean preoperative course of disease was 19.4 months (2-48 months). The patients were followed-up for a mean time of 12.4 months (8-20 months). The relationship between the origin of hallux ganglion and ankle joint was confirmed by ankle MRI and arthrography of ankle joint before the operation. During the operation, the joint capsule was explored, cleared and opened under the posterior ankle arthroscopy firstly, and then part of the tendon sheath of the forefoot was cleared and cut under the flexor hallucis longus tendoscope. American Orthopedic Foot and Ankle Society (AOFAS) forefoot rating scale and visual analogue scale (VAS) of pain were used to evaluate the clinical outcome before and after operation. Postoperative recurrence, perioperative and long-term complications were recorded spontaneously. Results: The cystic fluid signal of all patients could be traced from flexor hallucis longus tendon to the back of ankle joint by MRI before the operation. Intraoperative radiography showed that contrast media flowed from the back of ankle joint to the end of the hallux. The AOFAS score [M(Q1, Q3)] of the patients increased from 70.5(69.0, 87.8) before the operation to 100.0(85.8, 100.0) at the follow-up (P=0.002), and at the mean time, the VAS score decreased from 5.0(3.5, 6.0)to 0.5(0.0, 1.3) (P<0.001). No complications such as infection and recurrence occurred in all patients until the last follow-up. Conclusion: Posterior ankle arthroscopy combined with minimally invasive technique of tendoscope can treat ankle-origin flexor hallucis longus tendon ganglion effectively.

[Role of histone demethylase KDM6B in HBx-mediated podocyte-macrophage transdifferentiation].

Objective: To explore the expression of lysine (k)-specific demethylase 6B (KDM6B) in the renal tissues of hepatitis B virus-associated glomerulonephritis (HBV-GN) patients and human podocytes transfected with hepatitis B virus X (HBx) gene, and its role in HBx-mediated podocyte-macrophage transdifferentiation (PMT). Methods: Forty-eight patients diagnosed as HBV-GN by renal biopsy from 2013 to 2018 at the Shanghai Jiaotong University Affiliated First People's Hospital were included in this study. Thirty patients with primary glomerulonephritis (PGN) and fifteen patients with renal tumor were chosen as control group. The expression of KDM6B and macrophage marker F4/80 in renal tissues of HBV-GN patients was observed by immunofluorescence and immunohistochemistry. The association between kidney KDM6B levels and clinical features of HBV-GN patients was analyzed. The expression of KDM6B, F4/80, major histocompatibility complex (MHC)-Ⅱ and CD40 in the podocytes was detected by Western blotting. The contents of interferon-γ (IFN-γ) and interleukin-6 (IL-6) in the supernatant were determined by enzyme-linked immunosorbent assay (ELISA). The small interfering RNA of KDM6B (KDM6B siRNA) was used to silence the expression of KDM6B and the protein levels of KDM6B, F4/80 and tri-methylation of lysine 27 of histone H3 (H3K27me3) induced by HBx gene transfection were detected by Western blotting. Results: Renal KDM6B expression was significantly increased in HBV-GN patients compared to normal control (0.022±0.004 vs 0.006±0.002, P=0.006). There was no significant difference in the positive rate of KDM6B among different pathological types of HBV-GN (P=0.139). Moreover, co-expression of KDM6B and F4/80 could be observed in the podocytes of HBV-GN patients. Patients with estimated glomerular filtration (eGFR)<60 ml·min-1·(1.73 m2)-1or proteinuria ≥ 3.5 g/day had a significantly higher renal KDM6B expression compared to control groups (all P<0.05). In addition, the expression of KDM6B, F4/80, MHC-II and CD40 was significantly up-regulated in the podocytes transfected with HBx gene (all P<0.05). The content of IFN-γ and IL-6 in the supernatant was significantly increased (all P<0.05). After gene silencing of KDM6B, the expression of F4/80 induced by HBx in the podocytes was significantly down-regulated, while the level of H3K27me3 was significantly increased (both P<0.05). Conclusions: HBx could induce KDM6B expression in podocytes and initiate PMT, thereby involving in the dysfunction of immune microenviroment in the renal tissues of HBV-GN.

[Development of a hyperkalemia risk assessment model for patients with chronic kidney disease].

Objective: To investigate risk factors for hyperkalemia among chronic kidney disease (CKD) patients and establish a risk assessment model for predicting hyperkalemia events. Methods: Clinical data of CKD patients (stage 3 to 5) hospitalized between May 2017 and June 2020 from 14 hospitals were retrospectively collected and divided into training dataset and validation dataset through balanced random sampling. Multivariate logistic regression analysis was used to analyze risk factors for hyperkalemia in CKD patients and the factors were scored. Receiver operating characteristic (ROC) curve was plotted and the area under the curve (AUC) was calculated. Meanwhile, the cut-off value with the best sensitivity and specificity were used to verify the accuracy of the model in validation dataset. Results: A total of 847 CKD patients were enrolled and further divided into training dataset (n=675) and validation dataset (n=172). There were 555 males and 292 females, with a mean age of (57.2±15.6) years. Multivariate logistic regression analysis showed that age, CKD stage, history of heart failure, history of serum potassium ≥5.0 mmol/L, diabetes, metabolic acidosis, and use of medications that increase serum potassium levels were risk factors for causing hyperkalemia in patients with CKD. Risk assessment model was established based on these risk factors. The AUC of the ROC curve was 0.809. Using 4 as the cut-off value, the sensitivity and specificity for predicting hyperkalemia events reached 87.1% and 57.0%, respectively. Conclusion: The model established in the current study can be used for predicting hyperkalemia events in clinical practices, which offers a new way to optimize serum potassium management in patients with CKD.

Higher liver stiffness scores are associated with early kidney dysfunction in patients with histologically proven non-cirrhotic NAFLD.

AIM: The association between Liver fibrosis (LF), as assessed by either histology or Liver stiffness measurement (LSM), and the presence of Early kidney dysfunction (EKD) was investigated in this study, as was also the diagnostic performance of LSM for identifying the presence of EKD in patients with Non-alcoholic fatty liver disease (NAFLD). MATERIALS AND METHODS: A total of 214 adults with non-cirrhotic biopsy-proven NAFLD were recruited from two independent medical centres. Their histological stage of LF was quantified using Brunt's criteria. Vibration-controlled Transient elastography (TE), using M-probe (FibroScan®) ultrasound, was performed in 154 patients and defined as significant when LSM was≥8.0kPa. EKD was defined as the presence of microalbuminuria with an estimated glomerular filtration rate≥60mL/min/1.73 m2. Logistic regression modelling was used to estimate the likelihood of having EKD with NAFLD (LSM-EKD model). RESULTS: The prevalence of EKD was higher in patients with vs without LF on histology (22.14% vs 4.82%, respectively; P<0.001) and, similarly, EKD prevalence was higher in patients with LSM≥8.0kPa vs LSM<8.0kPa (23.81% vs 6.59%, respectively; P<0.05). The area under the ROC curve of the LSM-EKD model for identifying EKD was 0.80 (95% CI: 0.72-0.89). LF detected by either method was associated with EKD independently of established renal risk factors and potential confounders. CONCLUSION: LF was independently associated with EKD in patients with biopsy-proven NAFLD. Thus, TE-measured LSM, a widely used technique for quantifying LF, can accurately identify those patients with NAFLD who are at risk of having EKD.

[Analysis of needlestick injuries and depression among the female medical staff in a city district in 2018].

Objective: To investigate needlestick injuries and depression among the female medical staff in a city district. Methods: Using cross-sectional survey methods, female medical staff from selected sixteen hospitals voluntarily participated in questionnaire survey on APP of mobile phone. They Data were uploaded assessment platform, saved by Excel, and analysed by SPSS 22.0 statistical software. Results: The lasted year, Incidence of needlestick injuries was 11.28%. Incidence of needlestick injuries was respectively 5.2%, 9.1% and 16.3% in Class-I hospital, Second-class Hospital and three levels of hospitals(χ(2)=28.900, P<0.01). And 6.3% Resting no more than 10 days of Incidence of needlestick injuries was 12.5%, others was 6.3%.Night shift no more than 5 days of Incidence of needlestick injuries was 8.5%, others was 15.1%. The rate of not less than 14 of BDI was respectively 18.6%, 29.1% and 32.7% in Class-I hospital, Second-class Hospital and three levels of hospitals (χ(2)=23.843, P<0.01). In recent one year, the rate of medical staff who have taken annual leave less than 10 days and higher than 10 days scoring over 14 in the Beck(Beck depression inventory, BDI) survey are respectively 29.1% and 17.8% (χ(2)=10.710, P<0.01). In recent one year, the rate of medical staff who worked in night shift less than 5 days and higher than 5 days each month scoring over 14 in the Beck survey are respectively 23.5% and 31.7%(χ(2)=8.886, P<0.01). In recent one year, the rate of medical staff who had needle injury and non-needle injury scoring over 14 in the Beck survey are respectively 35% and 25%(χ(2)=4.782, P<0.05). Conclusion: The needle injury rate, high score rate in Beck survey among the medical staff increased when the level of the hospital they worked for increased. Medical staff should improve their method of rest.

Metastatic pattern and prognosis of gastrointestinal stromal tumor (GIST): a SEER-based analysis.

PURPOSE: This SEER-based study aimed to explore and analyze the relationship of metastasis of liver, lung and bone of GIST patients and their prognosis. METHODS: The data of GIST patients were from Surveillance, Epidemiology, and End Results (SEER) database from 2010 to 2015 and all the statistical analyses were conducted by statistical software package SPSS (Version 22.0). RESULTS: A total of 4224 GIST patients were identified, of which 388 (9.19%) patients with liver metastasis, 20 (0.47%) patients with bone metastasis and 32 (0.76%) patients with lung metastasis. There was no significant difference of risk of bone or lung metastasis between patients with and without liver metastasis (P = 0.935). The median overall survival of patients with liver, bone, or lung metastasis was, respectively, 49 months, 18 months, and 20 months, which were all shorter than that of patients without metastasis. The overall survival of patients with both liver and bone metastasis and those with metastasis of all three sites was not significantly different from that of patients with only liver metastasis. The multivariate analysis showed age of less than 65 years, female patients, married status and receiving surgery were all the beneficial factors for prognosis of GIST patients with liver metastasis. CONCLUSIONS: Patients with metastasis had a poorer prognosis than those without. Liver metastasis might have no relationship with bone or lung metastasis and liver might play a more dominant role than the other two sites in the prognosis of GIST patients with metastasis. So, more attention should be paid to liver status in diagnosis and treatment of GIST patients.

[The Occupational and Procreation Health of Road Sweeping Female Sanitation Workers in a District of a City].

Objective: To investigate occupational and procreation health of roadsweeping female sanitation workers in a city district. Methods: Using cross-sectional survey methods, all 303 the road sweeping female sanitation workers of one company were selected and interviewed face to face with questionnaires in june 2017. Data were analysed by SPSS22.0 statistical software. Results: The total prevalence of reproduction tract infection was 21.47%. The prevalence of uterine fibroids was 6.38%, and menstruation abnormal rate was 25.17%. The sanitation worker of menstruation abnormal rate was 31.03%, which were exposed low temperature, automobile exhaust and air contaminant at the same time, or else was 19.61%. There were statistical significance between them. (χ(2)=5.161, P<0.05) The sanitation worker of menstruation abnormal rate was 28.81%, which were not satisfied with women/s rights protection, or else was 11.67%. There were statistical significance between them. (χ(2)=7.435, P<0.01) . Conclusion: Female sanitation workers in the city district were exposed to many occupational hazards. The status of procreation health among road sweeping female sanitation workers in the city district is not optimistic. Occupational health protection and laborers's rights protection should be emphasized to improve health status of female sanitation workers.

[Changes of insulin-like growth factor 1 axis in chronic kidney disease and its role in skeletal muscle atrophy].

Objective: To investigate the changes of insulin-like growth factor 1 (IGF-1) axis in chronic kidney disease (CKD) and its role in skeletal muscle atrophy. Methods: A total of 240 patients with CKD stage 1-5 (without dialysis treatment) were included between August 2016 and February 2017. Serum IGF-1 and IGF-1 binding protein-3 (IGFBP-3) were measured using chemiluminescence, and the influencing factors of serum IGF-1 and IGFBP-3 were analyzed. Besides, male Sprague-Dawley rats (200-250 g) were randomly assigned to the sham-operated control group (Control, n=15) and the 5/6 nephrectomy group (n=15) as CKD animal model. The expressions of local IGF-1 and IGF-1 receptor (IGF-1R) of skeletal muscles were evaluated at the level of transcription and protein by real-time PCR and Western blotting. Results: There was no significant correlation between changes of serum IGF-1 level and estimated glomerular filtration rate (eGFR) (r=-0.066, P=0.307). However, after multivariable adjustment, serum IGFBP-3 increased with decreasing of eGFR (r=-0.181, P=0.005) in adult CKD patients. In multivariate analysis, age, eGFR, serum cholesterol level and 24 h urinary protein quantification were independent factors of serum IGFBP-3 in patients with CKD (R2=0.243, P<0.001). Animal experiments showed that the expression of IGF-1 mRNA and protein decreased in skeletal muscles of CKD rats. Expressions of IGF-1R mRNA and protein were slightly reduced and phosphorylation of IGF-1R was severely impaired in skeletal muscles of CKD rats. Conclusions: IGF-1 levels seem to be independent of renal function, but IGFBP-3 levels increased with decreasing of eGFR, which may cause a low affinity of IGF-1 with IGF-1R in skeletal muscles. Low affinity of IGF-1 with IGF-1R, as well as the decreasing of IGF-1 synthesis could lead to disorders of IGF-1R phosphorylation, and thus cause atrophy of skeletal muscles.

Remote ischemic preconditioning provides neuroprotection: impact on ketamine-induced neuroapoptosis in the developing rat brain.

OBJECTIVE: Previous studies have demonstrated that the commonly used anesthetic ketamine can acutely increase apoptosis and have long-lasting detrimental effects on cognitive function as the animal matures. Remote ischemic preconditioning (RIPC) has been confirmed to have a cerebral protective role in animal models of brain damage. The aim of this study was to investigate whether RIPC can protect the developing brain from anesthetic-induced neurotoxicity. MATERIALS AND METHODS: To investigate the protective properties of RIPC, 60 new-born Sprague-Dawley (SD) rats were randomly allocated into four groups: ketamine (20 mg/kg was diluted in saline, six times at an interval of 2 hours); RIPC (left hind row ischemia 5 min, reperfusion 5 min, a total of four cycles); ketamine + RIPC: RIPC was induced at postnatal day 5 and rats underwent the same treatment with the ketamine group after 48 hours; and saline (group vehicle). Neuronal apoptosis in the frontal cortex and hippocampal CA1 region was measured 24 h after treatment using immunohistochemistry of cleaved caspase-3. Learning and memory abilities were tested at the age of 60 days by Morris water maze test. RESULTS: The percentage of cleaved caspase-3 immunohistochemical staining positive cells in the ketamine + RIPC group showed a more marked decline in neuronal apoptosis of the CA1 region than that in the ketamine group (p < 0.05) but not in the CA1 region (p > 0.05). The mice exposed to RIPC alone showed no difference from the saline-treated mice. Moreover, RIPC significantly reversed the learning and memory deficits observed at 60 days of age. CONCLUSIONS: Our data indicate that RIPC treatment provides protection against ketamine-induced neuroapoptosis in the frontal cerebral cortex but not in the hippocampal CA1 region in developing rats and attenuates long-term behavioural deficits as the animals mature, suggesting a new possible strategy for neuroprotection.

Isolation and characterization of microsatellite markers for Dendranthema morifolium (Asteraceae) using next-generation sequencing.

Dendranthema morifolium (Asteraceae) is a perennial herbaceous plant native to China. A long history of artificial crossings may have resulted in complex genetic background and decreased genetic diversity. To protect the genetic diversity of D. morifolium and enabling breeding of new D. morifolium cultivars, we developed a set of molecular markers. We used pyrosequencing of an enriched microsatellite library by Roche 454 FLX+ platform, to isolate D. morifolium simple sequence repeats (SSRs). A total of 32,863 raw reads containing 2251 SSRs were obtained. To test the effectiveness of these SSR markers, we designed primers by randomly selecting 100 novel SSRs, and amplified them across 60 cultivars representing five different petal shape groups. Sixteen SSRs were polymorphic with the number of alleles ranging from 6 to 19, and their expected and observed heterozygosities ranging from 0.477 to 0.848, and 0.250 to 0.804, respectively. The polymorphism information content ranged from 0.459 to 0.854 and the inbreeding coefficient ranged from -0.119 to 0.759. An unweighted pair-group method arithmetic average analysis was performed to survey the phylogenetic relationships of these 60 cultivars and five clusters were identified. These markers can be used for investigating genetic relationships and identifying elite alleles through linkage and association analyses.

Isolation and characterization of microsatellite markers for Eucommia ulmoides (Eucommiaceae), an endangered tree, using next-generation sequencing.

Eucommia ulmoides Oliver, a single extant species of Eucommiaceae, is an endemic dioecious tree in China. The natural resources of E. ulmoides have rapidly declined in recent years because of the over-collection of its cortex. To design a suitable protection strategy, it is necessary to develop a set of molecular markers to investigate genetic diversity and population structure of E. ulmoides. Pyrosequencing of an enriched microsatellite library by Roche 454 FLX+ platform was used to isolate simple sequence repeats (SSRs) for E. ulmoides. A total of 1568 SSRs that contained enough flanking sequences for primer pair design were identified from 45,236 raw sequence reads. One hundred SSRs were randomly selected to design primer pairs and polymerase chain reaction was performed. Among these 100 tested primer pairs, 16 were polymorphic across 18 individuals from three E. ulmoides populations. The number of alleles ranged from 3 to 8, with an average of 5.1. The expected heterozygosity ranged from 0.110 to 0.830, with an average of 0.648, and the observed heterozygosity ranged from 0.111 to 0.833, with an average of 0.524. The inbreeding coefficient ranged from -0.349 to 0.547. This set of microsatellite markers could be valuable for landscape genetic structure assessment and molecular marker-assisted breeding in E. ulmoides.

[Expression of renal PLA2R in patients with idiopathic membranous nephropathy and its relationship with the curative effect of immunotherapy].

OBJECTIVE: To detect the expression of renal M-type phospholipase A2 receptor (PLA2R) in patients with idiopathic membranous nephropathy (IMN), and to explore the relationship between renal PLA2R and the curative effect of immunotherapy. METHODS: A total of 56 patients who were diagnosed as IMN from January 2012 to June 2014 in the department of nephrology in First People's Hospital, Shanghai Jiaotong University, were included in this study. The expression of renal PLA2R was detected by immumofluorescence assay. The IMN patients were treated with corticosteroids and cyclophosphamide, and the relationship between renal PLA2R and the curative effect of immunotherapy was observed. RESULTS: The ratio of PLA2R related IMN (renal PLA2R-positive) patients was 71.4%(40/56). The recovery conditions in proteinuria and serum albumin were better in the non-PLA2R related IMN group since 6 months after the treatment (P<0.05). The overall response rate in PLA2R related IMN group was 58.3%, 62.5% and 62.5% after 6, 9, 12 months, respectively. However, the overall response rate in non-PLA2R related IMN group almost reached 100% after treatment for 6 months. Compared with PLA2R related IMN group, the time which patients reached complete remission was significantly shorter in the non-PLA2R related IMN group [(5.4±3.5) vs (10.5±1.6) months, P<0.05]. CONCLUSIONS: Detection of renal PLA2R can be helpful to diagnose IMN. Non-PLA2R related IMN patients usually have a better curative effect of immunotherapy and a shorter time to onset of efficacy.

Isolation and characterization of microsatellite markers for Osmanthus fragrans (Oleaceae) using 454 sequencing technology.

Osmanthus fragrans (Oleaceae) is an evergreen shrub or small tree that grows in south China. In this study, Roche 454 FLX+ sequencing combined with the magnetic bead enrichment method was used to isolate microsatellite markers from the genome of O. fragrans. A total of 1471 microsatellites that contained enough flanking sequences for primer pair design were identified from 89,633 raw sequencing reads. One hundred primer pairs were randomly chosen to test primer amplification efficiency. Among these tested primer pairs, 20 yielded polymorphic amplification products across 16 individuals from the Albus, Luteus, and Aurantiacus groups. The number of alleles ranged from 2 to 6, with an average of 3.7. The observed heterozygosity ranged from 0 to 0.813, with an average of 0.460. Shannon's information index ranged from 0.463 to 1.707, with an average of 0.975. Six loci (Of 05, Of 06, Of 08, Of 12, Of 15, and Of 19) deviated significantly from Hardy-Weinberg equilibrium (P < 0.05), which was due to an excess of homozygotes or heterozygotes. Nine pairs of loci (Of 01 and Of 05; Of 04 and Of 05; Of 01 and Of 06; Of 04 and Of 12; Of 02 and Of 13; Of 04 and Of 13; Of 12 and Of 13; Of 04 and Of 19; Of 05 and Of 19) showed significant linkage disequilibrium, which indicated significant allelic association between the loci. This set of microsatellite markers will be valuable for molecular marker-assisted breeding in O. fragrans.

The abnormal expression level of microRNA in epithelial-mesenchymal transition of peritoneal mesothelial cells induced by high glucose.

OBJECTIVE: To determine the expression level of the microRNA in the process of epithelial-mesenchymal transition (EMT) of the peritoneal mesothelial cells (PMCs) induced by high glucose. MATERIALS AND METHODS: The PMCs were cultured using M199 medium with 10% fetal bovine serum, and the EMT was induced by D-glucose stimulation. Epithelial-mesenchymal transition was determined by changes in cell morphology and the expression levels of the EMT marker genes. Changes in cell morphology were observed by inverted microscope, and the expression levels of the EMT marker genes were determined by real-time PCR. The expression levels of the microRNA were detected by real-time PCR with microRNA-specific stem-loop structure primer. RESULTS: The PMCs changed to fusiformis following a high-glucose medium stimulated for 48 hours, and the EMT marker genes changed significantly, such as the decrease of E-cadherin and an increase of Vimentin (p < 0.01). These results proved the EMT had been induced by high-glucose. Applying real-time PCR with microRNA-specific stem-loop structure primer, miR-193a increased notably (p < 0.01), and miR-15a and let-7e decreased (p < 0.01), while miR-16 and miR-21 had no significant changes (p > 0.05). Most importantly, the increase of miR-193a was correlated with stimulus duration. CONCLUSIONS: MicroRNA with abnormal expression levels have a primary role in regulating the EMT of PMCs induced by high glucose.

Microinjection of salusin-beta into the nucleus tractus solitarii inhibits cardiovascular function by suppressing presympathetic neurons in rostral ventrolateral medulla in rats.

Salusin-beta is newly identified bioactive peptide of 20 amino acids, which is widely distributed in hematopoietic system, endocrine system, and the central nervous system (CNS). Although salusin-beta extensively expressed in the CNS, the central cardiovascular functions of salusin-beta are unclear. Our main objective was to determine the cardiovascular effect of microinjection of salusin-beta into the nucleus tractus solitarii (NTS) in anesthetized rats. Bilateral or unilateral microinjection of salusin-beta (0.94-94 microg/rat) into the NTS dose-dependently decreased blood pressure and heart rate. Bilateral NTS microinjection of salusin-beta (9.4 microg/rat) did not alter baroreflex sensitivity. Prior application of the glutamate receptor antagonist kynurenic acid (0.19 microg/rat, n=9) into the NTS did not alter the salusin-beta (9.4 microg/rat) induced hypotension and bradycardia. However, pretreatment with the GABA receptor agonist muscimol (0.5 ng/rat) within the rostral ventrolateral medulla (RVLM) completely abolished the hypotension (-14+/-5 vs. -3+/-5 mm Hg, P<0.05) and bradycardia (-22+/-6 vs. -6+/-5 bpm, P<0.05) evoked by intra-NTS salusin-beta (9.4 microg/rat). In addition, we found that vagotomy didn't influence the actions of salusin-beta (9.4 microg/rat) in the NTS. In conclusion, our present study shows that microinjection of salusin-beta into the NTS significantly produces hypotension and bradycardia, presumably by suppressing the activities of presympathetic neurons in the RVLM.

The deposition of Notch1 in hepatitis B virus-associated nephropathy and its role in hepatitis B virus X protein-induced epithelial-mesenchymal transdifferentiation and immunity disorder in renal tubular epithelial cells.

SUMMARY: Notch1 plays an important role in the regulation of immune responses and epithelial-mesenchymal transdifferentiation (EMT). Previous studies have observed inflammatory cell infiltration and tubulointerstitial fibrosis in the renal biopsies from patients with HBV-associated glomerulonephritis (HBV-GN). We hypothesized that Notch1 may be involved in the progression of HBV-GN. In this study, we evaluated the distribution of Notch1 in patients with HBV-GN. Our results showed that Notch1 was mainly distributed in renal tubules and the interstitial area, and the expression levels of Notch1 had a positive correlation with the renal tubular pathology. In this respect, we used human proximal tubular epithelial cells (HK-2) as target cells, which were transiently transfected with the hepatitis B virus X (HBx) gene using a eukaryotic vector. HBx expression resulted in significantly increased detection of Notch1, alpha-smooth muscle actin (α-SMA), major histocompatibility complex-II (MHC-II), CD40 and interleukin-4 (IL-4). At the same time, E-cadherin and interferon-γ (IFN-γ) expression levels were significantly inhibited. These HBx-induced phenotypes were exacerbated by upregulation of Notch1. Knock-down of Notch1 by specific shRNA caused decreases of α-SMA, MHC-II, CD40 and IL-4, and increases of E-cadherin and IFN-γ. These findings suggest that Notch1 is significantly associated with renal tubular and interstitial lesions. Notch1 can mediate HBx-induced EMT of HK-2 cells, promote HBx-induced increases in immune molecule expression and exacerbation of cytokine disorders, which may contribute to the progression of HBV-GN. Inhibitors of Notch1 signalling may be useful as new therapeutics for the treatment of HBV-GN.

Low adenosine triphosphate activity in CD4+ cells predicts infection in patients with lupus nephritis.

OBJECTIVES: The ImmuKnow (Cylex) assay has been reported to predict the risk of infection in some diseases; however, it is uncertain whether ImmuKnow can predict the risk of infection in lupus nephritis (LN) patients receiving immunosuppressive therapy. METHODS: The ImmuKnow Immune Cell Function Assay (Cylex, Inc., Columbia, MD, USA) was applied to measure the activity of CD4+ T cells, as a marker of global immune-competence. The correlation between changes in T cell activation and the relative risk of over-immunosuppression as well as infection was studied. The amount of adenosine triphosphate (ATP) produced by CD4+ T cells in response to phytohemagglutinin (PHA) was measured for 74 LN patients without infection, 22 LN patients with severe infection (i.e. required hospitalisation), and 28 healthy controls. RESULTS: No correlation was found between the ATP level and systemic lupus erythematosus (SLE) activity. The mean ATP level was significantly lower in LN patients with infection than that in healthy controls (p<0.01) and non-infected LN patients (p<0.01). The mean ATP level in non-infected LN patients was not significantly different compared to healthy controls. A cut-off ATP value of 300 ng/mL predicted infection in LN patients with a specificity of 77% and a sensitivity of 77%. Multi-variable partial correlation coefficient between the ATP assay and severe infection was r =-0.040, p<0.001; CRP was r=0.962, p<0.001. CONCLUSIONS: The ImmuKnow assay may be effective in identifying an increased risk of infection in LN patients but is not correlated with SLE activity. Combined CRP value will increase the diagnostic rate of severe infection in SLE. Larger studies are required to establish clinical advantages of this assay in SLE treatment.