[Clinical features and prognostic analysis of transarterial chemoembolization combined with targeted immunotherapy in the treatment of patients with hepatitis B virus-related intermediate-and advanced-stage hepatocellular carcinoma with secondary cholestasis].

Objective: To study the clinical features and prognostic impact of transarterial chemoembolization (TACE), immune checkpoint inhibitors (ICIs), and tyrosine kinase inhibitors (TKIs) combination therapy regimens in the treatment of patients with hepatitis B virus-related intermediate-and advanced-stage hepatocellular carcinoma with secondary cholestasis. Methods: Patients with HBV-related intermediate-and advanced-stage hepatocellular carcinoma (HBV) who visited the Affiliated Hospital of Xuzhou Medical University between January 1, 2020, and December 31, 2022, were enrolled. TACE+TKIs +ICIs combination therapy was used to treat all patients. The occurrence and factors influencing cholestasis, as well as the impact on prognosis after combined therapy, were analyzed. The measurement data were compared using a t-test and a non-parametric rank sum test. The count data was compared using the χ(2) test. The survival rates were compared using a log-rank test between different groups. Results: A total of 106 cases with HBV-related intermediate-and advanced-stage hepatocellular carcinoma were enrolled. The probabilities of secondary cholestasis within 3 and 6 months, 1, 2, and 3 years after TACE+ICIs+TKIs combination therapy were 9.4%, 12.3%, 14.2%, 24.5%, and 24.5%, respectively. Patients with secondary cholestasis had persistent symptoms and rapid progression. During the treatment course, the median survival time was significantly longer in patients with hepatocellular carcinoma without secondary cholestasis than that of patients with cholestasis (26.9 months vs. 13.7 months, respectively, P < 0.05). Secondary cholestasis, baseline aspartate aminotransferase, and prothrombin activity levels were independent risk factors that affected the survival and prognosis of patients treated with combination therapy. There was no statistically significant difference in the occurrence of other adverse reactions between the two groups with secondary and non-secondary cholestasis during the treatment course (47.5% vs. 43.3%, χ(2)=0.058, P = 0.810). Conclusion: TACE+ICIs+TKIs therapy combination is relatively common in the treatment of patients with HBV-related intermediate-and advanced-stage hepatocellular carcinoma with secondary cholestasis. Moreover, accelerated disease progression is an independent risk factor affecting the survival and prognosis of patients.

[Impact of different diagnostic criteria for assessing mild micro-hepatic encephalopathy in liver cirrhosis: an analysis based on a prospective, multicenter, real-world study].

Objective: To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test. Methods: This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ (2) test. A kappa test was used to compare the consistency between groups. Results: After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea (Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences (P < 0.001). Conclusion: The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.

[Research advances in imaging techniques related to the diagnosis of pneumoconiosis].

Occupational pneumoconiosis (hereinafter referred to as pneumoconiosis) is the most harmful occupational disease in China. According to the current standard GBZ 70-2015 Diagnosis of Occupational Pneumoconiosis, pneumoconiosis is mainly diagnosed and staged by high kilovolt or digital radiography. Chest radiography in pneumoconiosis is the most widely studied and mature imaging technique in the diagnosis of pneumoconiosis. However, this technique has some limitations in the screening of some early pneumoconiosis and occupational health examination, and there is a certain risk of missed diagnosis and misdiagnosis. With the continuous development of imaging examination technology, computed tomography, magnetic resonance imaging, positron emission tomography-computed tomography and artificial intelligence technology as auxiliary imaging examination methods have shown different diagnostic values in the research of auxiliary diagnosis of pneumoconiosis. This paper summarizes the advantages and problems in the application of various kinds of imaging techniques, which provides a direction for the future research of imaging techniques related to the diagnosis of pneumoconiosis.

[Establishment of morphological reference values for the differential count of white blood cells in peripheral blood smear, as well as nucleated cells and megakaryocytes in bone marrow smear].

Objective: To establish the morphological reference values for the differential count of white blood cells in peripheral blood smear as well as nucleated cells and megakaryocytes in bone marrow smear. Methods: From April 2012 to June 2020, 4 221 healthy donors for hematopoietic stem cell transplantation in Hebei Yanda Lu Daopei Hospital were selected. The median age was 36 (3-72) years old, including 2 520 males and 1 701 females. They were divided into four groups according to age: children group, with age≤14 years old [n=334, 11 (3-14) years old], youth group, with age >14 years old and <45 years old [n=2 855, 33 (15-44) years old], middle-aged adult group, with age ≥45 years old and < 60 years old [n=929, 49 (45-59) years old], and older adult group, with age ≥60 years old [n=103, 62 (60-72) years old]. Gender subgroups were established in each age group. According to different hematopoietic characteristics, the children group were divided into two subgroups: children group 1 [n=48, 6 (3-7) years old] and children group 2 [n=286, 11 (8-14) years old]. According to the clinical routine, 100 white blood cells in peripheral blood, 200 nucleated cells in bone marrow, and cell numbers/4.5 cm2 for megakaryocytes were classified and counted. The results of cell count in different age and gender groups were compared, and the reference values of morphological classification were established for different groups with statistical or clinical significance. Results: Due to the existence of statistically significant differences between children and adult groups and different gender subgroups in adults (all P<0.05), the reference values were established for children group and adult gender subgroups. The counts of segmented neutrophils and lymphocytes in peripheral blood were 46.65(43.97-49.32)% and 44.00(10.60-65.10)% in children group 1, 50.73(49.50-51.96)% and 39.55 (38.36-40.74)% in children group 2, and 57.00 (39.00-75.23) % and 33.00 (17.00-52.00) % in adult group, respectively. Bone marrow segmented neutrophils, orthochromatic erythroblasts, and mature lymphocytes were 11.54 (10.68-12.41)%, 14.20 (13.19-15.21)%, and 23.99 (22.06-25.92)% in children group 1, 12.50 (7.00-21.50)%, 15.00(9.50-25.50)%, and 21.02 (20.24-21.81)% in children group 2, 13.50 (7.50-21.00)%, 16.50 (10.50-26.00)%, and 15.50 (7.50-26.00)% in adult male group, and 14.50 (8.00-24.50)%, 14.50 (9.00-23.00)%, and 17.50 (8.50-29.00)% in adult female group, respectively. The myelopoiesis/erythropoiesis ratio in children group, adult male group and adult female group was 1.86∶1 (1.14∶1-3.23∶1), 1.96∶1 (1.12∶1-3.19∶1), 2.22∶1 (1.30∶1-3.69∶1), respectively. The numbers of granular megakaryocytes and thromocytogenic megakaryocytes were 138 (25-567) cells/4.5cm2 and 86 (13-328) cells/4.5 cm2 in children group, and 92 (13-338) cells/4.5 cm2 and 38 (3-162) cells/4.5 cm2 in adult group, respectively. Conclusion: The morphological reference values for the differential count of white blood cells in peripheral blood smear as well as nucleated cells and megakaryocytes in bone marrow smear are successfully established, which is helpful to improve the application of morphological examination in disease screening, diagnosis and monitoring.

[Multidisciplinary regenerative treatment and mechanisms for rescuing a severe calciphylaxis patient with human amnion-derived mesenchymal stem cells].

Calciphylaxis is a rare disease with severe pain and high-mortality due to cutaneous ischemic necrosis and infection that currently lacks proved effective therapies. The occurrence of calciphylaxis in end stage kidney disease (ESKD) patients is known as calcific uremic arteriolopathy (CUA), which is characterized histologically by dermal microvessel calcification, intimal fibroplasia and microthrombosis. Here we innovatively treated a severe CUA patient with human amnion-derived mesenchymal stem cells (hAMSCs). A 34-year-old uremic woman was presented with progressive, painful malodorous ulcers in buttocks and mummified lower limbs. Skin pathological features supported the diagnosis of calciphylaxis. The patient was refractory to conventional multidisciplinary symptomatic therapies. With the approval of our hospital ethics committee, she was treated with hAMSCs including intravenous and local intramuscular injection, and external application of hAMSC culture supernatant to the wound area. During 15-month follow-up, the patient had regeneration of skin and soft tissues, with improved blood biochemical, inflammatory, mineral and bone metabolic indices and immunoregulation effects. After 15-month hAMSC treatment, the score of pain visual analog scale (VAS) decreased from 10 to 0, Bates-Jensen wound assessment tool (BWAT) score decreased from 65 to 13, and wound-quality of life (Wound-QoL) questionnaire score decreased from 68 to 0. We propose that hAMSC treatment is promising for CUA patients. The therapy is potentially involved in the multiple beneficial effects of inhibiting vascular calcification, stimulating angiogenesis and myogenesis, modulating adverse inflammatory and immunologic responses, promoting re-epithelialization and restoring skin integrity.

Air quality improvement assessment and exposure risk of Shandong Province in China during 2014 to 2020.

As one of the most polluted provinces in China, air pollution events occur frequently in Shandong. Based on the hourly (or daily) concentrations of six air pollutants (PM2.5, PM10, O3, NO2, SO2 and CO), the situations of air quality improvement in three kinds of cities (key cities, coastal cities and general cities) are assessed comprehensively during 2014-2020. Contrary to the daily maximum 8-h average ozone (MDA8 O3), the annual average concentrations of other pollutants show the downward trends during 2014-2020. Therein, the improvement rates of annual average concentrations of air pollutants in key cities are highest. By 2020, the day proportions of O3 as the primary pollutant are up to 38% in three kinds of cities. Besides, due to the impact of COVID-19, the monthly average concentrations of PM2.5, PM10, NO2, SO2 and CO in February 2020 decrease by 32.1-49.5% year-on-year. There are still about 50% of population exposed to high-risk regions (R i > 2), which are mainly concentrated in main urban areas and industrial areas. Thus, the adjustment of industrial structure and energy composition in the context of carbon peak and carbon neutrality should be implemented in the future. Supplementary Information: The online version contains supplementary material available at 10.1007/s13762-022-04651-5.

Double Superconducting Dome and Triple Enhancement of T_{c} in the Kagome Superconductor CsV_{3}Sb_{5} under High Pressure.

CsV_{3}Sb_{5} is a newly discovered Z_{2} topological kagome metal showing the coexistence of a charge-density-wave (CDW)-like order at T^{*}=94 K and superconductivity (SC) at T_{c}=2.5 K at ambient pressure. Here, we study the interplay between CDW and SC in CsV_{3}Sb_{5} via measurements of resistivity, dc and ac magnetic susceptibility under various pressures up to 6.6 GPa. We find that the CDW transition decreases with pressure and experience a subtle modification at P_{c1}≈0.6-0.9 GPa before it vanishes completely at P_{c2}≈2 GPa. Correspondingly, T_{c}(P) displays an unusual M-shaped double dome with two maxima around P_{c1} and P_{c2}, respectively, leading to a tripled enhancement of T_{c} to about 8 K at 2 GPa. The obtained temperature-pressure phase diagram resembles those of unconventional superconductors, illustrating an intimated competition between CDW-like order and SC. The competition is found to be particularly strong for the intermediate pressure range P_{c1}≤P≤P_{c2} as evidenced by the broad superconducting transition and reduced superconducting volume fraction. The modification of CDW order around P_{c1} has been discussed based on the band structure calculations. This work not only demonstrates the potential to raise T_{c} of the V-based kagome superconductors, but also offers more insights into the rich physics related to the electron correlations in this novel family of topological kagome metals.

[Related factors for hyperkalemia and its recurrence in maintenance hemodialysis patients].

Objective: To investigate the prevalence of hyperkalemia and recurrent hyperkalemia in maintenance hemodialysis patients, and identify the related factors. Methods: The general information and clinical data of maintenance hemodialysis patients in the hemodialysis center of the First Affiliated Hospital of Nanjing Medical University from January to December 2020 were collected. According to the level of serum potassium, the patients were divided into normokalemia group and hyperkalemia group. Patients in the hyperkalemia group were further divided into single hyperkalemia group and recurrent hyperkalemia group according to the number of occurrences of hyperkalemia. Then, the prevalence of different serum potassium abnormalities and the related factors were explored. Results: Among the 352 patients included in the final analysis, 129 cases (36.6%) were in the normokalemia group [mean age: (62±15) years, 99 males] and 223 cases (63.4%) were in the hyperkalemia group [mean age: (60±14) years, 153 males]. Multivariate logistic regression analysis demonstrated that, compared with the normokalemia group, night-time dialysis (OR=4.012, 95%CI: 1.519-10.601, P=0.005), concurrent diabetes (OR=1.947, 95%CI: 1.148-3.304, P=0.013) and the number of serum potassium tested before hemodialysis (OR=1.561, 95%CI:1.292-1.885, P<0.001) were independent risk factors for hyperkalemia. Among 223 patients with hyperkalemia, 78 cases (35.0%) were in single hyperkalemia group and 145 cases (65.0%) were in recurrent hyperkalemia group. Multivariate logistic regression analysis demonstrated that, serum calcium (OR=21.885, 95%CI: 3.740-128.077, P=0.001), peak value of serum potassium before hemodialysis (OR=63.157, 95%CI: 25.265-157.876, P<0.001) and the number of serum potassium tested before hemodialysis (OR=1.814, 95%CI: 1.378-2.388, P<0.001) were the independent risk factors for the recurrence of hyperkalemia. Conclusions: The prevalence of hyperkalemia is high in maintenance hemodialysis patients, especially in those with diabetes or night-time dialysis. It is necessary to monitor serum potassium regularly. In addition, high serum calcium and peak value of serum potassium before hemodialysis are related factors for recurrent hyperkalemia.

[Association of urinary sodium excretion with fluid overload in patients with chronic kidney disease].

Objective: To investigate the relationship between urinary sodium excretion and fluid overload (FO) in non-dialysis patients with chronic kidney disease (CKD). Methods: Patients with CKD stage 1-4 who underwent bioelectrical impedance (BIA) in the Department of Nephrology, Jiangsu Province Hospital from December 2019 to January 2021 were recruited. All enrolled patients were categorized into two groups according to whether or not they develop FO. Further, clinical parameters were compared between the two groups. Spearman correlation analysis was used to investigate the association between over hydration/extracellular water (OH/ECW) and clinical characteristics. Multivariate logistic regression analysis was performed to evaluate the relationship between urinary sodium excretion and FO (FO was defined as OH/ECW≥7%). Results: A total of 385 patients with CKD stage 1-4 were finally included in the study, with a mean age of (46±15) years. There were 216 male cases (56.1%), and 150 cases (39.0%) existed FO. Spearman correlation analysis indicated that OH/ECW positively correlated with urinary sodium excretion (r=0.147, P=0.004), urinary protein excretion (r=0.555, P<0.001) and systolic blood pressure (r=0.241, P<0.001), but inversely related to estimated glomerular filtration rate (eGFR) (r=-0.111, P=0.030) and serum albumin (r=-0.659, P<0.001). After adjusting for confounding factors including age, systolic blood pressure, diabetes, urinary protein excretion, serum albumin, serum sodium, serum chlorine, urinary calcium excretion, urinary phosphorus excretion and use of diuretics, multivariate logistic regression analysis demonstrated that higher level of urinary sodium excretion was associated with increased risk of FO in patients with CKD (OR=1.005, 95%CI: 1.000-1.011, P=0.048). Conclusion: High urinary sodium excretion is independently associated with fluid FO in non-dialysis patients with CKD.

[Control study of chest CT imaging features of aluminosis and silicosis patients].

Objective: To understand the chest CT features of aluminosis caused by alumina and to improve the understanding of the imaging findings of alumina pneumoconiosis. Methods: The chest CT findings of 17 cases of alumina-induced pneumoconiosis and 30 cases of silicosis (the control group) diagnosed in Zibo Occupational Disease Prevention Hospital from April 2015 to July 2020 were analyzed retrospectively. The characteristics of fibrosis of the two kinds of pneumoconiosis and the incidence of size, density, distribution, tractive bronchiectasis, pleural thickening and interlobular septal thickening of pneumoconiosis nodules were compared. Results: Alumina pneumoconiosis showed nodules with thickened interlobular septal of 66.67% (12/18) , honeycomb lung of 22.22% (4/18) , ground glass shadow of 61.11% (11/18) , simple nodules of 11.11% (2/18) , and no fusion mass. In the control group, the long-line fibrosis of nodules with thickened interlobular septal were 16.67% (5/30) , 6.67% (2/30) with honeycomb lung and ground glass density shadow, 23.33% (7/30) with fusion mass and 53.33% (16/30) with simple nodule. There were significant differences in CT findings of nodules with thickened interlobular septal, ground glass density shadow, fused mass and simple nodules between the two groups (P<0.05) . The interstitial beaded nodules were seen in 18 cases of alumina pneumoconiosis, 50.00% (9/18) of them were beaded nodules, 61.33% (46/75) of low density nodules and 38.89% (7/18) of central lobular nodules were seen in alumina pneumoconiosis. The average width of nodules was (1.29±0.38) mm. Central lobular nodules were seen in all 30 cases of silicosis, 10.00% (3/30) were mainly beaded nodules, low density nodules were 36.29% (90/248) , and the average width diameter of nodules was (1.85±0.58) mm. There were significant differences between the two groups (P<0.05) . Alumina pneumoconiosis was often accompanied by traction bronchiectasis, pleural thickening and interlobular septal thickening (11, 18, 17 cases, 61.11%, 100.00%, 94.44%) , compared with the control group (9, 18, 18 cases, 30.00%, 60.00%, 60.00%) . The differences were statistically significant (P<0.05) . The maximum CT value of noncalcified mediastinal lymphnodes in alumina pneumoconiosis was (103.43±26.33) HU, which was higher than that of the control group[ (75.22±16.70) HU], and the difference was statistically significant (P<0.05) . Conclusion: Alumina pneumoconiosis chest CT shows slightly low-density beaded nodules, thickened interlobular septal, and pulmonary interstitial fibrosis of ground-glass shadows, mostly combines with stretched bronchiectasis, thickened pleura, and mediastinum increased lymph node density.

Ancestry Inference Using the ForenSeqTM DNA Signature Prep Kit.

ABSTRACT: Objective To evaluate the effect of 56 ancestry informative single nucleotide polymorphism （aiSNP） genetic markers in the ForenSeqTM DNA Signature Prep Kit on ancestry inference. Methods A total of 85 samples from five populations including Hebei Han population, Inner Mongolia autonomous region Mongolian population, Tibet autonomous region Tibetan population, Xinjiang Uygur autonomous region Uygur population and Nigerian population were collected. The library was constructed with the ForenSeqTM DNA Signature Prep Kit and sequencing was performed based on the MiSeq FGx Forensic Genomics System. Using universal analysis software （UAS） of ForenSeqTM, principal component analysis （PCA）, Structure and likelihood ratio method was used on the genotyping data of 56 aiSNP markers, respectively, and the genetic relationships between populations and inference of the origin of ancestors were analyzed. Results Among the five populations tested, the four ethnic populations in China （Hebei Han population, Inner Mongolia autonomous region Mongolian population, Tibet autonomous region Tibetan population and Xinjiang Uygur autonomous region Uygur population） could be significantly distinguished from Nigerian population. Xinjiang Uygur autonomous region Uygur individuals were shown as having mixed origins of ancestors and could be distinguished from the other three Chinese populations. However, the other three populations in China （Hebei Han population, Inner Mongolia autonomous region Mongolian population and Tibet autonomous region Tibetan population） could not be effectively distinguished by the system. Conclusion The 56 aiSNP markers in the ForenSeqTM DNA Signature Prep Kit can make accurate ancestry inference from the intercontinental level, but it is not yet able to distinguish between Chinese subpopulations.

Full Sibling Testing Based on NGS-SNP Genotyping Method and IBS Strategy.

ABSTRACT: Objective To evaluate the effectiveness of single nucleotide polymorphism （SNP） genoty-ping in combination with identity by state （IBS） strategy in full sibling testing. Methods Thirty-five blood samples were collected from a four-generation family. Ninety autosomal SNPs were genotyped using Precision ID Identity Panel. The distribution of IBS scores for full siblings and other relationships were calculated and compared. The relationships were determined using Fisher discriminant function and threshold method, respectively. Results Based on family members and previous research, 44, 30, 111, 71 and 1 000 pairs of full siblings （FS）, grandparent-grandchild （GG）, uncle/aunt-nephew/niece （UN）, first cousins （FC） and unrelated individuals （UI） were obtained, respectively. The average IBS scores were 148, 130, 132, 124 and 120, respectively. Except for the GG and UN pairs, the distribution differences among the other relationships had statistical significance （P<0.05）. The false rates of Fisher discriminant function to determine relationships were 1.3%, 22.3%, 17.0% and 38.7% for FS, GG, UN and FC, respectively. Based on the simulation data, the thresholds t1=128 and t2=141 were recommended to determine full sibling relationships （the false rate ≤0.05%）. Conclusion The 90 SNP genetic markers included in the Precision ID Identity Panel meet the testing requirements for full sibling relationships. The threshold method based on IBS has a relatively lower false rate and is more flexible.

The association of time of emergency surgery - day, evening or night - with postoperative 30-day hospital mortality.

It is unclear whether the time of day for emergency surgery is associated with postoperative mortality. We assessed this association in 9319 patients who had emergency surgery as their first surgery at the Jewish General Hospital, Montreal, QC, Canada from April 2010 to March 2015. Of the 7362 (80%) patients with complete data, 168 (2.3%) died within 30 postoperative days. There was no significant association of time of day with postoperative mortality, with adjusted OR (95%CI) of 1.61 (0.96-2.72) for night vs. day, p = 0.07; 1.29 (0.78-2.13) for night vs. evening, p = 0.33; and 1.26 (0.89-1.78) for evening vs. day, p = 0.20. Studies of more patients and more factors, with longer follow-up, should be carried out to exclude important associations of time of emergency surgery with postoperative mortality and morbidity.

[Effect and related mechanism of microRNA-181 attenuates oxidized low density lipoprotein induced vascular endothelial cell injury].

Objective: To observe the expression level of microRNA-181 (miR-181) and importin-α3 in oxidized low density lipoprotein (ox-LDL) induced vascular endothelial cell injury models, and explore the effect and mechanism of miR-181 on endothelial cell injury. Methods: Human vein endothelial cell line CRL-1730 were cultured and vascular endothelial cell injury model was established by intervention with ox-LDL. The cells were divided into control group (intervened by double distilled water), low-dose group (intervened by 10 µg/ml ox-LDL) and high-dose group (intervened by 20 µg/ml ox-LDL). In addition, cells of low-dose group were divided into miR-181 mimic group (miR-181 mimic was transfected) and mimic control group (miR-181 mimic control was transfected). Cell viabilities, mRNA and protein expression level of interleukin-6 (IL-6), miR-181, importin-α3, and nuclear transcription factor-κB (NF-κB) were measured by methyl thiazolyl tetrazolium (MTT), real-time PCR and Western blot, respectively. Results: (1) The cell viabilities in low-dose group and high-dose group were lower than control group (0.207±0.012 and 0.204±0.007 vs. 0.323±0.018, all P<0.01). The relative IL-6 mRNA expression in low-dose group and high-dose group were higher than control group (1.24±0.16 and 1.36±0.23 vs. 0.22±0.03, all P<0.01). The relative miR-181 mRNA expression in low-dose group and high-dose group were lower than control group (0.91±0.11 and 0.88±0.07 vs. 2.20±0.13, all P<0.01). The relative importin-α3 mRNA expression in low-dose group and high-dose group were higher than control group (1.23±0.22 and 0.55±0.03 vs. 0.44±0.06, all P<0.01). The relative NF-κB mRNA expression in low-dose group and high-dose group were higher than control group (1.67±0.34 and 0.41±0.11 vs. 0.11±0.04, all P<0.01). The relative importin-α3 protein expression in low-dose group and high-dose group were higher than control group (1.44±0.23 and 1.31±0.22 vs. 0.29±0.08, all P<0.01). The relative NF-κB protein expression in low-dose group and high-dose group were higher than control group (0.43±0.05 and 0.37±0.04 vs. 0.16±0.03, all P<0.01). (2)The cell viabilities in miR-181 mimic group was higher than in mimic control group (0.262±0.008 vs. 0.211±0.021, P<0.01). The relative miR-181 mRNA expression level in miR-181 mimic group was higher than in mimic control group (4.23±0.34 vs. 0.88±0.16, P<0.01). The relative importin-α3 mRNA expression level in miR-181 mimic group was lower than in mimic control group (0.24±0.03 vs. 1.08±0.13, P<0.01). The relative NF-κB mRNA expression level was lower in miR-181 mimic group than in mimic control group (0.13±0.03 vs. 0.51±0.06, P<0.01). The relative importin-α3 protein expression level was lower in miR-181 mimic group than in mimic control group (0.34±0.06 vs. 1.67±0.26, P<0.01). The relative NF-κB protein expression level was lower in miR-181 mimic group than in mimic control group (0.43±0.02 vs. 1.53±0.36, P<0.01). Conclusions: Lower miR-181 expression but higher importin-α3 and its downstream NF-κB signaling are associated with ox-LDL induced vascular endothelial cell injury and up-regulation of miR-181 could alleviate ox-LDL induced vascular endothelial cell injury possibly via importin-α3/NF-κB pathway.

Riga-Fede disease in a child.

Riga-Fede disease (RFD) was initially described as a benign ulceration of the lingual frenum or tongue of infants, most commonly resulting from repetitive trauma by natal teeth. However, similar clinical manifestations can also be found in children with repetitive tongue-thrusting habits or certain neurological disorders. This report describes an unusual case of RFD in a 6-year-old Chinese boy. The lesion appeared as an ulcer on the anterior ventral surface of the child's tongue, and was caused by a repetitive tongue-thrusting habit. After eliminating the source of repetitive trauma and starting concurrent use of therapeutic drugs for 1 week, the lesion healed completely.

Purification and characterization of a potential antifungal protein from Bacillus subtilis E1R-J against Valsa mali.

In order to identify the antagonistic substances produced by Bacillus subtilis E1R-J as candidate of biocontrol agents for controlling Apple Valsa Canker, hydrochloric acid precipitation, reverse phase chromatography, gel filtration, and ion exchange chromatography were used. The purified fraction EP-2 showed a single band in native-polyacrylamide gel electrophoresis (native-PAGE) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Fraction EP-2 was eluted from native-PAGE and showed a clear inhibition zone against V. mali 03-8. These results prove that EP-2 is one of the most important antifungal substances produced by B. subtilis E1R-J in fermentation broth. SDS-PAGE and Nano-LC-ESI-MS/MS analysis results demonstrated that EP-2 was likely an antifungal peptide (trA0A086WXP9), with a relative molecular mass of 12.44 kDa and isoelectric point of 9.94. The examination of antagonistic mechanism under SEM and TEM showed that EP-2 appeared to inhibit Valsa mali 03-8 by causing hyphal swelling, distortion, abnormality and protoplasts extravasation. Inhibition spectrum results showed that antifungal protein EP-2 had significantly inhibition on sixteen kinds of plant pathogenic fungi. The stability test results showed that protein EP-2 was stable with antifungal activity at temperatures as high as 100 °C for 30 min and in pH values ranging from 1.0 to 8.0, or incubated with each 5 mM Cu(2+), Zn(2+), Mg(2+), or K(+). However, the antifungal activity was negatively affected by Proteinase K treatment.

Plasma FGF23 levels and heart rate variability in patients with stage 5 CKD.

UNLABELLED: Fibroblast growth factor 23(FGF23) is a bone-derived hormone which regulates mineral homeostasis but may also have a role in cardiovascular disease. Here, we found that higher plasma FGF23 was independently associated with decreased heart rate variability in stage 5 CKD patients and parathyroidectomy may reverse these abnormal indicators. INTRODUCTION: Lower heart rate variability (HRV) in patients with chronic kidney disease (CKD) compared with healthy controls is associated with increased risk of cardiovascular disease (CVD). Higher levels of plasma FGF23 also predict higher risk of CVD. Here, we aimed to evaluate the relationship between plasma FGF23 levels and HRV in patients with stage 5 CKD and to investigate longitudinal changes of them together with the correlation between their changes in two severe secondary hyperparathyroidism (SHPT) subgroups with successful parathyroidectomy (PTX) and persistent SHPT. METHODS: This cross-sectional study included 100 stage 5 CKD patients, 78 controls, and a prospective study in two PTX subgroups classified as successful PTX (n = 24) and persistent SHPT (n = 4) follow-up. Blood examination and 24-h Holter monitoring for HRV were measured. RESULTS: Most HRV indices were lower in stage 5 CKD patients than in healthy controls, and plasma FGF23 levels were higher. In multivariate stepwise regression models, levels of plasma FGF23 and serum parathyroid hormone (PTH) were correlated with HRV. The successful PTX subgroup had significant improvements over baseline in HRV indices. Persistent SHPT subgroup had numerically similar changes in HRV indices. However, plasma FGF23 levels decreased in both subgroups. CONCLUSIONS: Plasma FGF23 levels were higher in CKD patients than in controls, much higher in patients with severe SHPT. FGF23 was independently associated with decreased HRV in stage 5 CKD. Successful PTX may reverse these abnormal indicators and contribute to decreases in the risk of cardiovascular disease.

An enzyme-linked immunosorbent assay for detection of pyrene and related polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons (PAHs) can form DNA-binding compounds that show genotoxicity and carcinogenicity. Pyrene, as a PAH, was covalently linked to carrier protein bovine serum albumin and ovalbumin. A monoclonal antibody (McAb) was produced that showed high cross-reactivity values with chrysene (169.73%), benzo[a]pyrene (693.34%), benzo[a]anthracene (16.36%), and indeno[1,2,3-cd]pyrene (40.96%) and showed no significant cross-reactivity values with other homologues (<0.1%). A competitive enzyme-linked immunosorbent assay (ELISA) was developed for detection of pyrene and some homologues in water samples. The detection limit of the assay was 65.08 pg ml(-1). The average recoveries of PAHs from tap water, lake water, and mineral water were 99.13, 99.74, and 99.19%, respectively, indicating that matrices of water samples do not interfere with the assay. The results demonstrated that the developed ELISA seems to be a potential method for monitoring of pyrene and some homologous PAHs in water samples.

Purification, characterization, and heterologous expression of an antifungal protein from the endophytic Bacillus subtilis strain Em7 and its activity against Sclerotinia sclerotiorum.

An antifungal protein exhibiting a high activity against Sclerotinia sclerotiorum in vivo was purified by ammonium sulfate precipitation, hydrophobic chromatography, and gel filtration chromatography from the culture filtrate of the endophytic Bacillus subtilis strain Em7. The protein was characterized as a ß-1,3-1,4-glucanase according to amino acid analysis, and showed excellent properties in thermal stability and acid resistance. At the same time, the antifungal protein was cloned and heterologously expressed in Escherichia coli BL21. The recombinant protein was purified and showed similar enzymatic properties to the native protein, exhibiting strong inhibitory activity against S. sclerotiorum. This shows that the ß-1,3-1,4-glucanase may play a very important role in B. subtilis Em7 biocontrol function. In addition, many physiochemical properties of the native and purified recombinant protein were compared, including the effect of pH, temperature, metal cations, substrate specificity, and kinetic parameters. All parameters were similar between the native and recombinant purified protein, indicating that the purified recombinant protein has potential for industrial applications.

Total parathyroidectomy with autotransplantation for a rare disease derived from uremic secondary hyperparathyroidism, the uremic leontiasis ossea.

SUMMARY: We described six uremic leontiasis ossea (ULO) patients who underwent total parathyroidectomy with autotransplantation. ULO demonstrated more a systemic disease than a simple craniofacial deformation. The surgery seemed an effective treatment to alleviate secondary hyperparathyroidism and to improve patients' quality of life. ULO may have a high postoperative recurrence tendency. INTRODUCTION: ULO is a rare disease derived from uremic secondary hyperparathyroidism (SHPT). Previous studies mostly focused on the craniofacial deformations. This study aims to investigate the systemic features of the disease and the surgical outcomes. METHODS: The present study retrospectively assessed six ULO patients who underwent total parathyroidectomy (TPTX) with autotransplantation (AT). Follow-up data were recorded. The follow-up status was considered as "effectiveness" if serum intact parathyroid hormone (iPTH) levels were <150 pg/mL in the first 3 days after surgery, or as "recurrence" if serum iPTH gradually increased >300 pg/mL during follow-up in patients whose status was initially considered as "effectiveness". RESULTS: Craniofacial deformations, short stature, thoracocyllosis, spine malformations, osteodynia, and muscle weakness were observed in all patients. Abnormal pulmonary functions were observed in five patients. After surgery, one patient died from respiratory failure. Surgery was effective in the remaining five patients with relieved osteodynia and stopped craniofacial deformation. A mean follow-up of 7.6 (4 to 12) months was available. Three patients suffered from recurrence of hyperparathyroidism originating from autografts. CONCLUSIONS: Our data suggests that ULO is not only a simple disease with craniofacial malformations but is a severe systemic disease leading to increased surgical risks. TPTX with AT seems an effective treatment to relieve SHPT and to improve quality of life. ULO may have a high postoperative recurrence tendency.