[Value of constructing a non-invasive diagnostic model based on serum heme oxygenase-1 and glucose regulatory protein 78 for non-alcoholic fatty liver disease].

Objective: To analyze the clinical application value of serum heme oxygenase (HO)-1expression level in non-alcoholic fatty liver disease (NAFLD) and, based on that, establish a diagnostic model combined with glucose regulatory protein 78 (GRP78) so as to clarify its diagnostic effectiveness and application value. Methods: A total of 210 NAFLD patients diagnosed by abdominal B-ultrasound and liver elastography were included, and at the same time, 170 healthy controls were enrolled. The general clinical data, peripheral blood cell counts, and biochemical indicators of the research subjects were collected. The expression levels of HO-1 and GRP78 were detected using an enzyme-linked immunosorbent assay. Multivariate analysis was used to screen independent risk factors for NAFLD. Visual output was performed through nomogram diagrams, and the diagnostic model was constructed. Receiver operating characteristic curve (ROC), calibration curve, and decision curve analysis (DCA) were used to evaluate the diagnostic effectiveness of NAFLD. Measurement data were analyzed using a t-test or Mann-Whitney U rank sum test to detect data differences between groups. Enumeration data were analyzed using the Fisher's exact probability test or the Pearson χ(2) test. Results: Compared with the healthy control group, the white blood cell count, aspartate aminotransferase (AST), alanine aminotransferase, gamma-glutamyl transferase (GTT), fasting blood glucose (Glu), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), serum HO-1, and GRP78 levels were significantly increased in the NAFLD group patients (P < 0.05). Binary logistic analysis results showed that AST, TG, LDL-C, serum HO-1, and GRP78 were independent risk factors for NAFLD (P < 0.05). A nomogram clinical predictive model HGATL was established using HO-1 (H), GRP78 (G) combined with AST (A), TG (T), and LDL-C (L), with the formula P=-21.469+3.621×HO-1+0.116 ×GRP78+0.674×AST+6.250×TG+4.122 ×LDL-C. The results confirmed that the area under the ROC curve of the HGATL model was 0.965 8, with an optimal cutoff value of 81.69, a sensitivity of 87.06%, a specificity of 92.82%, a P < 0.05, and the diagnostic effectiveness significantly higher than that of a single indicator. The calibration curve and DCA both showed that the model had good diagnostic performance. Conclusion: The HGATL model can be used as a novel, non-invasive diagnosis model for NAFLD and has a positive application value in NAFLD diagnosis and therapeutic effect evaluation. Therefore, it should be explored and promoted in clinical applications.

Image findings of tendon sheaths affected by diffuse tenosynovial giant cell tumors of the skull base.

OBJECTIVE: This study investigated radiographic images and the differential diagnosis of intracranial diffuse tenosynovial giant cell tumor (D-TGCT) in order to better understand the disease and improve the rate of preoperative diagnosis. PATIENTS AND METHODS: Images and clinical data of patients with D-TGCT were retrospectively analyzed. Routine Computer Tomography (CT), routine Magnetic Resonance Imaging (MRI), and contrast-enhanced MRI were performed for nine cases. Susceptibility-weighted imaging (SWI) was also performed for one case. RESULTS: We reviewed nine patients (6 males and 3 females) aged between 24 and 64 years, with a mean age of 47.33 ± 14.92 years. The most frequent complaints were hearing loss (5/9, 55.6%), pain (4/9, 44%), masticatory symptoms (2/9, 22.2%), and mass (4/9, 44.4%), with a mean duration of 22 ± 21.43 months. All cases were centered on the base of the skull, and showed hyper-density soft-tissue mass with osteolytic bone destruction on CT. The tumor signal mainly showed iso-intensity or hypo-intensity on T1WI compared with that in the brain parenchyma in all patients. On T2WI, nine lesions mainly showed hypo-intensity. Among these nine lesions, three displayed cystic region showing hyper-intensity on T2WI and hypo-intensity on T1WI (Figure 2A, 2B) in the lesion. Nine lesions showed hypo-intensity on DWI sequences. SWI images presented low signal in two cases, showing the "flowering effect". Nine patients showed heterogeneous enhancement, and two patients had meningeal thickening. CONCLUSIONS: Intracranial D-TGCT is extremely rare, but must be differentiated from other tumors. Osteolytic bone destruction in the area of the skull base with hyper-density soft-tissue mass and hypo-intensity on T2WI images are indicative of D-TGCT.

[The correlation of CD49d expression pattern with molecular genetics and hotspot gene mutants in patients with chronic lymphocytic leukemia].

Objective: To explore the correlation of CD49d expression patterns with molecular genetics and hotspot gene mutants in patients with chronic lymphocytic leukemia. Methods: The expression of CD49d was detected by flow cytometry and grouped into homogeneous, bimodal, negative and positive expression. Panel fluorescence in situ hybridization (FISH) was used for molecular genetics analysis and next-generation sequencing (NGS) was conducted for gene mutation detection. Results: There were 43 patients (23.89% ) with positive CD49d expression, 137 patients (76.11% ) with negative CD49d expression, 96 patients (53.33% ) with homogeneous CD49d expression and 84 patients (46.67% ) with bimodal CD49d expression. Compared with patients in the CD49d negative group, patients in the CD49d positive group had higher Rai stage (P=0.048) and higher proportion of spleen enlargement (P=0.030) . Compared with patients with homogeneous expression of CD49d, patients with bimodal expression of CD49d had a higher proportion of spleen enlargement (P=0.009) . The expression rate of 11q22- in bimodal CD49d(-) group was significantly higher than that in homogeneous CD49d(-) group (24.29% vs 10.45% , P=0.043) . The incidence of +12 in homogeneous CD49d group was higher than that in bimodal CD49d group (16.67% vs 5.95% , P=0.035) . The incidence of +12 in homogeneous CD49d(+) group was higher than that in bimodal CD49d(-) group (17.24% vs 4.29% , P=0.045) . The incidence of +12 in homogeneous CD49d(-) group was higher than that in bimodal CD49d(-) group (16.42% vs 4.29% , P=0.024) . BIRC3 mutation rate in CD49d positive group was higher than that in CD49d negative group (11.63% vs 2.92% , P=0.037) . Conclusion: There were significant correlations between CD49d and 11q22-, +12 and BIRC3 gene mutation. Patients with bimodal CD49d were more correlated with poor prognosis indexes.

[Study of cytogenetics and molecular biology in typical and atypical immunophenotypic chronic lymphocytic leukemia].

Objective: To analyze the differences in immunophenotype, cytogenetics, and molecular biology between typical and atypical immunophenotype chronic lymphocytic leukemia (CLL) , and explore the correlation of cytogenetic anomalies with gene mutations. Methods: This study included 488 patients diagnosed in the First Affiliated Hospital of Nanjing Medical University between November 2014 and May 2021. Of these, 382 patients scored 4-5 points, which was typical CLL (tCLL) , and 106 scored 3 points, which was atypical CLL (aCLL) as per the Royal Marsden Hospital Immunomarker Integral System. Peripheral blood cells were collected for immunophenotype by multiparameter flow cytometry in 488 patients, fluorescence in situ hybridization (FISH) was employed to detect cytogenetic anomalies in 359 patients, and gene mutations were detected by next-generation sequencing (NGS) in 330 patients. Results: The positive rates of CD10, CD22, CD49d, CD81, and FMC7 were significantly higher in the aCLL compared with the tCLL group (P=0.020, P<0.001, P<0.001, P=0.027, and P<0.001, respectively) , while the positive rates of CD5, CD23, CD148, and CD200 were lower in the former compared to the latter (P<0.001, P=0.017, P=0.041, and P<0.001, respectively) . aCLL exhibited a higher frequency of trisomy 12 and lower frequency of del (13q14) compared to the tCLL group (P<0.001 and P<0.001, respectively) . Moreover, aCLL patients also showed a higher incidence of NOTCH1 mutations than the tCLL patients (P=0.038) , while no statistically significant differences in other gene mutations occurred between the two groups. No significant differences in overall survival (OS) and treatment-free survival (TFS) occurred between aCLL and tCLL using Kaplan-Meier analysis (P>0.05) . Conclusion: aCLL has characteristic immunophenotype, cytogenetic, and somatic mutation that differ from tCLL, and this can provide reliable information for the diagnosis and differential diagnosis between the two groups.

[Progress in the diagnosis of minimal hepatic encephalopathy].

Hepatic encephalopathy (HE) is a common serious complication of liver cirrhosis, with sudden onset, indicating a poor prognosis in patients with chronic liver disease. Minimal hepatic encephalopathy (MHE) is an early stage of HE with no apparent symptoms, but it shows abnormal results in neuropsychological and/or neurophysiological tests. MHE affects patients' quality of life, employability, driving ability, and has a high risk of developing overt hepatic encephalopathy (OHE). This article aims to explore various diagnostic methods, strengthen the routine work of clinicians in diagnosis and treatment, and develop an effective MHE screening protocol.

[The expressional characteristics and diagnostic values of TRBC1 in mature T-cell lymphoma].

Objective: To investigate the expression characteristics of TRBC1 protein in mature T-cell lymphoma (TCL) , and compare with T-cell receptor (TCR) -Vß repertoire analysis and TCR gene rearrangement results, to explore the value of TRBC1 in the diagnosis of TCL. Methods: The expression of TRBC1 was detected by multi-parameter flow cytometry in 30 cases of TCL, 40 cases of normal controls and 50 cases of patients without T lymphocyte proliferative diseases (non-TCL) admitted to the Department of Hematology, The First Affiliated Hospital of Nanjing Medical University. The diagnostic value of TCRVß repertoire analysis, TCR gene rearrangement and TRBC1 restricted expression detection in TCL was evaluated. Results: The positive rates of CD4(+)T and CD8(+)T cell subsets TRBC1 in normal control group were (39.6±6.5) % and (39.3±4.4) %. The positive rates of CD4(+)T and CD8(+)T cell subsets TRBC1 in non-TCL were (39.1±3.8) % and (36.0±8.4) %. All 30 cases of TCL were CD3(+)TCRγδ(-), and the positive rate of TRBC1 was >92.3% or <12.7%. All cases showed restrictive expression pattern (monoclonal expression) , which was significantly different from those of the normal control and the non-TCL cases (P<0.001) . In terms of the diagnostic performance of T cell clonality, the sensitivity of TRBC1 was 100%, the positive detection rate of TCR gene rearrangement was 92.8%, and the sensitivity of TCRVß detection was 94.1%. Kappa test showed high consistency among the three detective methods. Conclusion: Multi-parameter flow cytometry detection of TRBC1 expression level can quickly and efficiently diagnose mature T-cell lymphoma, which has good clinical application value.

[Clinical and genetic features of seven children with MYH9-related disease].

Objective: To summarize and analyze of the clinical and genetic characteristics of children with nonmuscle myosin heavy chain 9 (MYH9)-related disease (MYH9-RD). Methods: To screen the patients who were first diagnosed as "chronic/refractory immune thrombocytopenia (ITP) " from April 2016 to May 2019 in Beijing Children's Hospital by genetic and clinical examinations, then the clinical manifestation, laboratory examination and genetics results of 7 children diagnosed with MYH9-RD were collected and summarized retrospectively. Results: Among 7 children diagnosed with MYH9-RD, 3 were males and 4 females. The age of onset was 1.25 (0.41-6.16) years. The course of disease was 2.16 (0.41-8.59) years. The automatic platelet count was (9 (5-30))×109/L. All the cases were found with giant platelets under microscope,and the manual platelet count was (70 (30-100))×109/L. Four cases had skin hemorrhage or epistaxis and 3 cases had no bleeding. All 7 patients had received first-or second-line therapy of ITP, of whom 1 case received splenic embolization, and all the treatments mentioned above were ineffective. Finally, it was confirmed that all 7 patients had heterozygous missense mutations of MYH9 gene by next generation sequencing (NGS), including 2 pedigrees and 5 sporadic cases. Four sporadic mutations occurred in N-terminal globular head domain (HD), and 1 sporadic case with p.D1424N mutations occurred in the C-terminal tail domain (TD). One of the pedigrees also had p.D1424N mutation. The other familial case had a novel variant with one missense variant p.A44D caused by the c.131C>A transition. One of the two p.R702 mutations had kidney damage, and several relatives of the new p.A44D mutations had deafness. Conclusions: In this study, the spontaneous mutations of seven MYH9-RD were common, and all patients were misdiagnosed as ITP, whereas the bleeding was mild and immunotherapy was ineffective. The suspected disease can be identified earlier by manual visual platelet volume and count, which can be confirmed by genetic testing. It is more important to monitor the development of other organs damage instead of thrombocytopenia. For cases with p.R702 mutations the doctor should be aware of kidney damage, and for the cases with novel mutations p.A44D the doctor should be aware of hearing loss.

[Establishment and application of HPLC-MS/MS method for detecting m6A level in RNA].

Objective: Explore the establishment of a fast, stable and sensitive high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for detecting the level of m6A modification in RNA and its application. Methods: The degree of m6A in RNA can be expressed as the ratio of m6A and adenosine (A) in concentration, which can be determined by ESI source positive ion multiple reaction monitoring (MRM) mode. The established method was verified by analyzing three quality control samples (m6A: 4, 40, 400 nmol/L; A: 40, 400, 4 000 nmol/L) with three different concentrations of low, medium, and high. The method was used to detect the degree of m6A in RNA from mouse spleen T cells treated in different ways. The t test was used to compare the differences between the two groups of data. Results: The established method had a good Linearity (R2>0.99) in a range of 1-500 nmol/L for m6A and 10-5 000 nmol/L for A. The limit of detection (LOD) was 1 nmol/L for m6A and 10 nmol/L for A. The recoveries were between 98.9% and 116.5%. The intra-day (n=5) RSDs and the inter-day (n=15, 5 days) RSDs were 2.4%-9.5% and 4.4%-9.6%, respectively. And this method was used to detect the degree of m6A in the RNA from mouse spleen T cells cultured in different conditions. The results showed that the m6A modification level in the RNA of primary CD8+T cell was 0.271 5±0.017 9, and the m6A modification level in the RNA of primary CD8+T cell with IL-27 was 0.251 7±0.015 0, indicating that primary CD8+T cells have a higher level of RNA methylation. Conclusion: This research has established a fast, simplemethylation degree in RNA with HPLC-MS/MS. This method is easy to be popularized and is suitable for the detection of large quantity of samples, and of great significance in analyzing the relationship between methylation and diseases.

Associations among temporal and large artery abnormalities on vascular ultrasound in giant cell arteritis.

Objectives: Giant cell arteritis (GCA) can manifest in cranial and/or extracranial arteries. We investigated the distribution of affected arteries on vascular ultrasound (VUS) among patients with new-onset or prior-onset GCA.Method: We retrospectively studied patients with either new-onset or prior-onset GCA and an abnormal VUS, from 2013 to 2017. Trained vascular technologists imaged the bilateral temporal arteries and carotid, axillary, and subclavian arteries. Vascular medicine physicians interpreted the images. Vasculitis-related abnormalities in individual vessels and their distribution (temporal artery, large artery, or both) were evaluated. Phi coefficients (φ) and Fisher's exact test were used to assess correlations among individual abnormal arteries.Results: Among 66 GCA patients, 28.8% had prior-onset GCA (median duration 17.8 months). Acute arteritis on VUS was observed in the majority of patients with both new-onset (72.3%) and prior-onset GCA (68.4%); the remainder had hyperechoic wall thickening without acute arteritis. Involvement of the temporal arteries only (45.5%) or large arteries only (34.8%) was more common than involvement of both (19.7%); this finding was similar in new-onset and prior-onset GCA. There were moderate positive correlations among temporal artery branches (φ = 0.51-0.58, p < 0.003) and among axillary and subclavian arteries (φ = 0.51-0.77, p < 0.003), and moderate negative correlations between abnormalities in the temporal and large arteries (φ = -0.46 to -0.58, p < 0.003).Conclusion: On VUS, vasculitis-related abnormalities in the temporal arteries only or large arteries only were more common than concurrent temporal and large artery abnormalities in patients with both new-onset GCA and prior-onset GCA.

Screening a Lactobacillus plantarum strain for good adaption in alfalfa ensiling and demonstrating its improvement of alfalfa silage quality.

AIM: To screen a lactic acid bacteria (LAB) strain for good adaption in alfalfa ensiling and to evaluate its effects on alfalfa ensiling. METHODS AND RESULTS: Lactobacillus plantarum (LP) strain ZZU203 was selected due to its superior ability to acidify structural carbohydrate metabolite and alfalfa powder. Alfalfa was ensiled with LP FG1 (from a commercial inoculant), LP ZZU203, or a no-additive control (CK) for 60 d. The two silages with LP additives significantly (P < 0·05) increased lactic acid (LA) and acetic acid (AA) concentrations compared to CK silage. The pH, ammonia nitrogen (NH3 -N) and cellulose concentrations, and the numbers of bacilli, coliform, aerobic bacteria and Clostridium decreased significantly (P < 0·05). The dry matter, LA, AA and water-soluble carbohydrate concentrations of ZZU203 silage were significantly (P < 0·05) higher, whereas the NH3 -N and the neutral detergent fibre concentrations, and the aerobic bacterial and Clostridium counts were significantly (P < 0·05) lower than in FG1 silage. CONCLUSION: ZZU203 obtained by the screening method of this study can improve the quality of alfalfa silage. SIGNIFICANCE AND IMPACT OF THE STUDY: The screening method designed for alfalfa silage conditions can effectively screen out LAB with potential application value in alfalfa silage.

Antidiarrheal effect of bioactivity-guided fractions and bioactive components of pomegranate (Punica granatum L.) peels.

BACKGROUND: Pomegranate peels have been widely used to treat diarrhea in China. The antidiarrheal activities of aqueous extracts of pomegranate peels have been evaluated. However, there have not been any bioactivity-guided fractionation studies on the antidiarrheal effect to identify the bioactive components of the extract. METHODS: Bioactivity-guided fractionation of an aqueous extract of pomegranate peels was performed using different solvents of increasing polarity, generating fractions dissolved in ethyl acetate, n-butyl alcohol, and the residual fraction. The principal chemical composition of the active fraction was analyzed by HPLC/ESI-MS. KEY RESULTS: Fecal frequencies revealed that only the ethyl acetate fraction possessed significant antidiarrheal activity. Furthermore, administration of the ethyl acetate fraction at 100, 200, and 400 mg/kg significantly reduced gastrointestinal transit in charcoal meal tests in mice. It also significantly inhibited castor oil-induced enteropooling compared to control animals. Histopathological analysis revealed that small intestine lesions of mice treated with the ethyl acetate fraction were alleviated compared to those in mice treated with castor oil. The ethyl acetate fraction was found to be composed mainly of punicalagin, corilagin, and ellagic acid, and a combination of these compounds could mediate the antidiarrheal activities. CONCLUSION AND INFERENCES: Our study describes the protective effects of pomegranate peels against castor oil-induced diarrhea. The findings showed that the ethyl acetate fraction was the active fraction of pomegranate peels, of which punicalagin, corilagin, and ellagic acid were responsible for the antidiarrheal effect of aqueous extracts.

Inhibition of Nkcc1 promotes axonal growth and motor recovery in ischemic rats.

Bumetanide is a selective inhibitor of the Na+-K+-Cl--co-transporter 1(NKCC1). We studied whether bumetanide could affect axonal growth and behavioral outcome in stroke rats. Adult male Wistar rats were randomly assigned to four groups: sham-operated rats treated with vehicle or bumetanide, and ischemic rats treated with vehicle or bumetanide. Endothelin-1 was used to induce focal cerebral ischemia. Bumetanide administration (i.c.v.) started on postoperative day 7 and continued for 3 weeks. Biotinylated dextran amine (BDA) was injected into the right imotor cortex on postoperative day 14 to trace corticospinal tract (CST) fibers sprouting into the denervated cervical spinal cord. Nogo-A, NKCC1, KCC2 and BDNF in the perilesional cortex and BDA, PSD-95 and vGlut1 in the denervated spinal cord were measured by immunohistochemistry and/or Western blot. Behavioral outcome of rats was assessed by the beam walking and cylinder tests. The total length of CST fibers sprouting into the denervated cervical spinal cord significantly increased after stroke and bumetanide further increased this sprouting. Bumetanide treatment also decreased the expressions of NKCC1 and Nogo-A, increased the expressions of KCC2 and BDNF in the perilesional cortex and enhanced the synaptic plasticity in the denervated cervical spinal cord after cerebral ischemia. The behavioral performance of ischemic rats was significantly improved by bumetanide. In conclusion, bumetanide promoted post-stroke axonal sprouting together accompanied by an improved behavioral outcome possibly through restoring and maintaining neuronal chloride homeostasis and creating a recovery-promoting microenvironment by overcoming the axonal growth inhibition encountered after cerebral ischemia in rats.

Wildlife Plague Surveillance Near the China-Kazakhstan Border: 2012-2015.

Plague is a zoonotic disease caused by the bacterium Yersinia pestis. This pathogen can be transmitted by fleas and has an enzootic cycle, circulating among small mammals, and occasionally epizootic cycles, infecting other species. In China, infected wild rodents are primarily reservoirs of Y. Pestis and are related to human infection (Int. J. Infect. Dis., 33, 2015 and 67; BMC Microbiol., 9, 2009 and 205). Because shepherd dogs prey on and eat rodents (e.g. marmots and mice), they are valuable sentinel animals for plague serosurveillance in endemic disease foci, although their infections are usually asymptomatic (Vet. Microbiol., 172, 2014 and 339).

Forced limb-use enhanced neurogenesis and behavioral recovery after stroke in the aged rats.

Constraint-induced movement therapy (CIMT) after stroke enhances not only functional reorganization but also structural plasticity of the brain in the adult rats. We examined whether forced limb-use which mimicked CIMT could influence ischemia-induced neurogenesis, apoptosis and behavioral recovery in the aged rats. Aged rats were divided into a sham group, an ischemia group, and an ischemia group with forced limb-use. Focal cerebral ischemia was induced by injection of endothelin-1. Forced limb-use began on post-stroke day 7 by fitting a plaster cast around the unimpaired upper limbs of rats for 3 weeks. Behavioral recovery was evaluated by tapered/ledged beam-walking test on postoperative day 32. The expression of doublecortin, neuronal nuclei, glial fibrillary acidic protein and Iba-1 were measured by single or double immunohistochemistry, and apoptosis was measured by TdT-mediated dUTP-biotin nick-end labeling (TUNEL) assay. The production of neuroblasts in the subventricular zone (SVZ) was significantly increased after stroke. Forced limb-use enhanced the proliferation of newborn neurons in the SVZ, as well as increased the long-term survival of newborn neurons. Furthermore, forced limb-use suppressed apoptosis and improved the motor functions after stroke in the aged rats. Forced limb-use exerted few effects on inflammation. Neither the number nor dendritic complexity of newborn granule cells in the hippocampus was affected by forced limb-use. Forced limb-use is effective in enhancing neurogenesis and behavioral recovery after stroke even in the aged rats.

Inverse correlation between CD4+ CD25high CD127low/- regulatory T-cells and serum immunoglobulin A in patients with new-onset ankylosing spondylitis.

This study investigated the role of regulatory T (T(reg)) cells in patients with new-onset, treatment-naïve ankylosing spondylitis (AS). Levels of CD4(+)CD25(high)CD127(low/-) T(reg) cells in the peripheral blood of 14 AS patients and 18 age-matched healthy volunteers were investigated by flow cytometry and correlations with serum levels of immunoglobulin A (IgA) and AS activity, as assessed by the Bath AS Disease Activity Index (BASDAI), were analysed. The number of peripheral blood CD4(+)CD25(high)CD127(low/-) T(reg) cells in AS patients was found to be significantly lower than in healthy controls and was inversely correlated with serum IgA levels. There was no significant correlation between CD4(+)CD25(high)CD127(low/-) T(reg) cell numbers and BASDAI scores. It is concluded that CD4(+)CD25(high)CD127(low/-) T(reg) cells may play a role in the pathogenesis and activity of AS.

High-sensitivity photovoltaic responses in manganite-based heterojunctions on Si substrates for weak light detection.

We have fabricated and characterized a weak light photodetector in a heterojunction composed of manganite La0.4Ca0.6MnO3 and n-type Si. High-sensitivity photoresponse properties were investigated. The responsivities of open-circuit photovoltage and short-circuit photocurrent reach ∼1000 V/mJ and ∼30 A/mJ, respectively, without any amplification bias under irradiation by 20-ps-wide and 355, 532, and 1064-nm-wavelength laser pulses in nanojoule to microjoule order. The present results demonstrate that the manganite-based heterojunction on Si substrate has potential applications in weak light detection from ultraviolet to near-infrared light.