Pulmonary Epstein-Barr virus-associated smooth muscle tumor after kidney transplantation: two case reports with review of differential diagnosis.

Pulmonary nodules are a common complication in solid organ transplant recipients, and may have various underlying causes, with Epstein-Barr virus-associated smooth muscle tumor (EBV-SMT) being one of them. Given the rarity of this entity, we describe the diagnosis and therapeutic interventions for post-transplant EBV-SMT in two individuals. Both cases involved female patients who were diagnosed with multiple pulmonary nodules 60 months and 116 months, respectively, after receiving living-related kidney transplantation. Pathological examination revealed a spindle cell tumor, with immunophenotype and EBV in situ hybridization supporting the diagnosis of EBV-SMT. After diagnosis, these two patients underwent intervention by decreasing their intake of immunosuppressants. As of the latest follow-up, the patients' lesion size remained stable, and their overall condition was favorable. We also reviewed literature about the morphological and molecular pathological features of EBV-SMT and highlighted the diagnosis and differential diagnosis of pulmonary spindle cell lesions especially in the setting of immunosuppression.

Sodium butyrate attenuate hyperglycemia-induced inflammatory response and renal injury in diabetic mice.

The activation of the monocyte-macrophage system and the damage to the renal and pancreatic tissue are common complications in patients with diabetes induced by hyper-glycemia. This study aimed to evaluate the effect and mechanism of butyrate (NaB), a metabolite of intestinal flora, on inhibiting the inflammatory response of human monocyte-macrophages (THP-1 cells) induced by high glucose and the damage of pancreatic and renal tissue in diabetic mice. The results showed that high concentration glucose significantly up-regulated the expressions of IL-1ß, TNF-α, and NLRP3 in THP-1 cells and mouse spleen, and that NaB could inhibit the overexpression of those genes. The abundance of Beclin-1, LC3B and reactive oxygen species (ROS) in THP-1 cells is increased due to the high glucose concentration, and NaB can inhibit the genes responsible for upregulating the expression. In diabetic mice, vacuolar degeneration of renal tubules was observed. Then we observed that some of the epithelial cells of the renal tubules were exfoliated and some formed tubules. NaB could alleviate these pathological lesions, but NaB cannot alleviate pancreatic injury. Our results indicated that NaB could be used for the prevention and adjuvant treatment of diabetic kidney injury.

The characteristics of a 12-lead electrocardiogram measuring premature ventricular contractions originating from the tricuspid annulus using the clock position method.

OBJECTIVE: This study aimed to analyze the conventional surface electrocardiogram (ECG) characteristics of premature ventricular contractions (PVCs) originating from the tricuspid annulus and to investigate the efficacy of locating their origins according to ECG results. METHODS: Eight patients who underwent radiofrequency ablation in the First Hospital of Shanxi Medical University (China) were included in the study. Pace mapping (PM) was used to analyze the characteristics of the PVCs originating from the tricuspid annulus recorded via 12-lead body surface ECGs. RESULTS: An R-wave was found in leads I, V5 , and V6 . The QRS wave was narrower when the PVCs originated from the septum and shifted in lead V3 (R-wave amplitude/S-wave amplitude in the precordial lead-1). The QRS wave was broadest when the PVCs originated from the 7 to 9 o'clock position. The augmented vector left lead showed RS, QS, or RSR-type waves with a low amplitude when the PVCs originated from the upper part of the annulus. When the PVCs originated from the lower part of the annulus, the augmented vector right lead reflected multidirectional and QS-type waves. CONCLUSION: The ECG-lead characteristics related to the origin of PVCs in the tricuspid annulus indicate some level of significance and can be used to formulate a specific diagnosis.

Expert Consensus on Pathological Diagnosis of Intrahepatic Cholangiocarcinoma (2022 version).

Intrahepatic cholangiocarcinoma (iCCA) can originate from the large bile duct group (segment bile ducts and area bile ducts), small bile duct group (septal bile ducts and interlobular bile ducts), and terminal bile duct group (bile ductules and canals of Hering) of the intrahepatic biliary tree, which can be histopathological corresponding to large duct type iCCA, small duct type iCCA and iCCA with ductal plate malformation pattern, and cholangiolocarcinoma, respectively. The challenge in pathological diagnosis of above subtypes of iCCA falls in the distinction of cellular morphologies, tissue structures, growth patterns, invasive behaviors, immunophenotypes, molecular mutations, and surgical prognoses. For these reasons, this expert consensus provides nine recommendations as a reference for standardizing and refining the diagnosis of pathological subtypes of iCCA, mainly based on the 5th edition of the World Health Organization Classification of Tumours of the Digestive System.

A standardized pathological proposal for evaluating microvascular invasion of hepatocellular carcinoma: a multicenter study by LCPGC.

BACKGROUND AND AIMS: Microvascular invasion (MVI) is a key pathological factor that severely affects the postoperative prognosis of patients with hepatocellular carcinoma (HCC). However, no MVI classification schemes based on standardized gross sampling protocols of HCC are available at present. METHODS: 119 HCC specimens were sampled at multiple sites (3-, 7-, and 13 points) for the optimum MVI detection rate. 16,144 resected HCCs were graded as M0, M1 or M2 by adopting three-tiered MVI grading (MVI-TTG) scheme based on the seven-point sampling protocol (SPSP). Survival analyses were performed on 2573 patients to explore the advantages of MVI-TTG. RESULTS: The MVI detection rate determined by SPSP was significantly higher than that determined by the 3-point sampling method (34.5% vs. 47.1%, p = 0.048), but was similar to that determined by the 13-point sampling method (47.1% vs. 51.3%, p = 0.517). Among 16,144 resected HCCs, the proportions of M0, M1 and M2 specimens according to SPSP were 53.4%, 26.2% and 20.4%, respectively. Postoperative survival analysis in 2573 HCC patients showed that the 3-year recurrence rates in M0, M1 and M2 MVI groups were 62.5%, 71.6% and 86.1%, respectively (p < 0.001), and the corresponding 3-year overall survival (OS) rates were 94.1%, 87.5% and 67.0%, respectively (p < 0.001). M1 grade was associated with early recurrence, while M2 grade was associated with both early and late recurrence. MVI-TTG had a larger area under the curve and net benefit rate than the two-tiered MVI grading scheme for predicting time to recurrence and OS. CONCLUSIONS: SPSP is a practical method to balance the efficacy of sampling numbers and MVI detection rates. MVI-TTG based on SPSP is a better prognostic predictor than the two-tiered MVI scheme. The combined use of SPSP and MVI-TTG is recommended for the routine pathological diagnosis of HCC.

Thyroid-Like Intrahepatic Cholangiocarcinoma: Report of a Case and Review of the Literature.

Intrahepatic cholangiocarcinoma is a relatively uncommon malignant neoplasm. We recently encountered an unusual case of intrahepatic cholangiocarcinoma that histologically resembled a thyroid carcinoma. A thorough review of the English literature revealed only 2 similar cases that have been previously reported. Immunohistochemical studies are imperative to confirm the diagnosis of cholangiocarcinoma and to exclude the possibility of metastatic thyroid carcinoma and other malignancies with thyroid-like features.

Identification of New Epididymal Luminal Fluid Proteins Involved in Sperm Maturation in Infertile Rats Treated by Dutasteride Using iTRAQ.

BACKGROUND: Spermatozoa become mature and acquire fertilizing capacity during their passage through the epididymal lumen. In this study, we identified new epididymal luminal fluid proteins involved in sperm maturation in infertile rats by dutasteride, a dual 5α-reductase inhibitor, in order to provide potential epididymal targets for new contraceptives and infertility treatment. METHODS: Male rats were treated with dutasteride for 28 consecutive days. We observed the protein expression profiles in the epididymal luminal fluids in infertile and normal rats using isobaric tags for relative and absolute quantitation (iTRAQ) technique. The confidence of proteome data was validated by enzyme-linked immunosorbent assays. RESULTS: 1045 proteins were tested, and 23 of them presented different expression profiling in the infertile and normal rats. The seven proteins were down-regulated, and 16 proteins were up-regulated. Among the seven proteins which were significantly down-regulated by dutasteride in the epididymal luminal fluids, there were three ß-defensins (Defb2, Defb18 and Defb39), which maybe the key proteins involved in epididymal sperm maturation and male fertility. CONCLUSIONS: We report for the first time that dutasteride influences the protein expression profiling in the epididymal luminal fluids of rats, and this result provides some new epididymal targets for male contraception and infertility therapy.

Clinicopathological features of cryoglobulinemic glomerulonephritis associated with HBV infection: a retrospective analysis of 8 cases in China.

AIMS: We retrospectively analyzed clinicopathologic features of 8 cases of hepatitis B virus-associated glomerulonephritis with hyaline thrombi, to confirm the diagnosis of cryoglobulinemic glomerulonephritis (CRYGN) associated with HBV infection. METHODS: Retrospective analysis was carried out with demographic information, clinical manifestations, laboratory parameters, pathological and prognostic features. RESULTS: The median age of 8 patients was 30.5 years (range, 21-75 years), including 6 males and 2 femles (M:F = 3:1). One patient had Raynaud's syndrome. Cryoglobulin testing was performed in 4 cases of our series, and 3 cases had elevated cryocrit (>256). Serum C4 decreased in all detected cases. Histopathologically, all cases showed hyaline thrombi occluded in capillary lumina; Co-deposit of IgG, IgM, IgA, Fib, C3d, C4d, C1q, HBsAg and HBcAb were identified in hyaline deposit/hyaline thrombi with polyclonal Igκ and Igλ staining. Ultrastructural examination confirmed the hyaline thrombi to be huge electron-dense bodies, which were a homogeneous texture. CONCLUSIONS: The results suggest that 8 cases in the series are CRYGN associated with HBV infection. The incidence of CRYGN associated HBV was extremely low. Our series suggested that prognosis of CRYGN associated HBV was better in patients with mild symptoms, but it was poor in elder patients with severe vasculitis.

[Inhibitory effect of dutasteride on the expressions of epididymal Claudin1 and ß-catenin in male rats].

OBJECTIVE: To explore the molecular mechanism of dutasteride inhibiting fertility by studying its effects on the expressions of the epididymal epithelial junction proteins Claudin1 and ß-catenin in rats. METHODS: Sixteen 3-month-old SD male rats were equally divided into an experimental and a negative control group to be treated intragastrically with dutasteride at 40 mg/kg per day and the same dose of solvent, respectively, for 14 consecutive days. Then, the sperm motility and morphology of the rats were detected by computer-assisted sperm analysis, the serum levels of testosterone (T) and dihydrotestosterone (DHT) measured by ELISA, changes in the tight junction of epididymal cells observed under the transmission electron microscope, the protein and gene expressions of Claudin1 and ß-catenin determined by RT-PCR and immunohistochemistry, and the conception rate of the mated female rats calculated. RESULTS: Dutasteride significantly suppressed the serum DHT level, sperm motility, and fertility of the rats (P <0.05). Interspaces between epididymal epithelial cell tight junctions were observed, the volume of epididymal fluid obviously increased, and the expressions of Claudin1 and ß-catenin gene and protein remarkably downregulated in the experimental rats (P <0.05). CONCLUSION: Dutasteride can significantly inhibit the fertility of male rats by reducing the serum DHT level, suppressing Claudin1 and ß-catenin expressions, and damaging epididymal epithelial cell junctions.

Increased expression of cannabinoid receptor 1 in the nucleus accumbens core in a rat model with morphine withdrawal.

Relapse is a major clinical problem and remains a major challenge in the treatment of drug addiction. There is strong evidence that the endocannabinoid system of the nucleus accumben core (NAcc) is involved in drug-seeking behavior, as well as in the mechanisms that underlie relapse to drug use. To reveal the mechanism that underlies this finding, we examined the expression pattern of the cannabinoid receptor 1 (CB1-R) in the NAcc of SD rats that had been undergoing morphine withdrawal (MW) for 1 day, 3 days and 3 weeks (acute, latent and chronic phases, respectively). Morphine exposure induced conditioned place preference (CPP) in rats. Significant increase of CB1-R expression in NAcc was observed in animals in the 1 day, 3 days and 3 weeks morphine withdrawal compare to the control group. Immunofluorescence labeling showed axonal fibers or terminals by fluorescence microscope observation. Immunoelectron microscopy detection showed silver-gold particles located in the presynaptic membranes that mainly give rise to symmetrical synapses. Quantitative electron microscopy showed an increase in number of CB1-R-positive terminals in the morphine withdrawal groups and the number of immunogold particles was significantly increased at these inhibitory terminals. We also confirmed that infusions of the CB1-R antagonist rimonabant into the NAcc attenuated the CPP during morphine withdrawal. Our present data have thus indicated that increasing pattern of CB1-R expression in the NAcc during above morphine withdrawal phases, which might underlie the relapse associated drug seeking behavior after morphine withdrawal.

The continual presence of C3d but not IgG glomerular capillary deposition in stage I idiopathic membranous nephropathy in patients receiving corticosteroid treatment.

BACKGROUND: Pathologic diagnosis of stage I idiopathic membranous nephropathy (MN-I) requires electron microscopy or immunohistochemistry that shows a glomerular capillary staining pattern of IgG and C3. However, it is not uncommon that renal biopsy did not obtain sufficient material for electron microscopy and that IgG and C3 staining in glomeruli largely lost at biopsy due to corticosteroid treatment. Since C3d is one of the final degradation products of C3 that is more stable in vivo, we determine if C3d staining could be used as a novel immunohistochemical marker for MN-I. METHODS AND RESULTS: 74 MN-I patients with electron microscopy proven MN-I were examined by immunoperoxidase staining of C3d. Intensive C3d staining was present in glomerular capillary like the staining pattern of IgG and C3 in MN-I. Importantly, in 40 MN-I patients who underwent corticosteroid treatment at biopsy the intensity and glomerular capillary pattern of C3d staining remained largely intact while the staining for IgG had substantially reduced and the pattern of glomerular capillary staining became unrecognizable. CONCLUSIONS: C3d glomerular capillary staining may be a novel marker for pathologic diagnosis of MN-I that is continuously present at biopsy in patient who has received corticosteroid treatment. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2120780075734479.

[An experimental study on the safety of intratracheal drug administration].

OBJECTIVE: To explore whether injury and repair occur in the trachea and the lung after intra-tracheal administration of different drugs. METHODS: Wistar rats were randomly divided into 5 groups, a normal group, a blank control (BC) group, a normal saline (NS) group, a lidocaine (LD) group and an amikacin (AK) group. For the latter 3 groups, normal saline, lidocaine and amikacin were injected into trachea by needle puncture. Scanning electron microscope was used to observe the ultra-structural changes of the epithelium, and the percentage of the area of damage (PAD) in tracheal mucosa was calculated. Moreover, pathological changes of the mucous membrane of bronchioles and alveolar epithelial cells were also examined, and the degree of lung pathology was semi-quantified. RESULTS: Two hours after the injection of the 3 drugs, derangement and edema of the cilia were evident by scanning electron microscopy. The PAD of the NS group, the LD group and the AK group were (94.2 ± 3.2)%, (93.1 ± 3.0)% and (95.5 ± 1.8)%, respectively; all being significantly higher than that of the BC group (1.3 ± 0.3)%. For the NS group and the LD group, the PAD decreased significantly after 24 h, which were (73.7 ± 7.8)% and (81.0 ± 4.6)% respectively, and returned to normal at 48 h and 96 h. While for the AK group, the damage began to improve at 72 h [PAD (62.1 ± 5.2)%], and recovered at 96 h. Airway epithelial derangement and cell edema in the alveoli and the bronchioles also occurred 2 h after drug injection, and inflammatory cell infiltration became evident at 24 h. At this time, the score of pathology was 1.80 ± 0.84, 2.60 ± 0.55 and 2.80 ± 0.45 for the NS group, the LD group and the AK group, respectively; all being higher than that of the BC group (0). These pathological changes recovered totally after 72 h for the NS and the LD groups, and 96 h for the AK group. CONCLUSIONS: Intra-tracheal administration of normal saline, lidocaine and amikacin in rats led to reversible airway mucosal and lung tissue damages.

[Pathological features and ultrastructure of N-methyl-N-nitrosourea (MNU)-induced thymic lymphomas in mice].

The objective of study was to investigate the origin and to classify the subtype of N-methyl-N-nitrosourea (MNU)-induced thymic lymphomas in mice. Histopathologic, immunohistochemical and ultrastructural studies were performed to analyze the pathological features of the neoplasms. The results showed that the thymus in all cases became totally replaced by sheets of cells of the lymphoid series. All the tumors coexpressed CD3 and TdT. Transmission electron microscopic study showed the plasma membranes of malignant lymphoma cells were smooth. The nuclear profiles were usually regular, with varying percentage of convoluted nuclei. Few cell organoids were observed in cytoplasm. In conclusion, all the MNU-induced tumor classified by histopathologic, immunohistochemical and ultrastructural studies as precursor lymphoblastic lymphoma that were unquestionably related to the thymus origin and T-cell lineage.

A phiC31 integrase-mediated integration hotspot in favor of transgene expression exists in the bovine genome.

phiC31 integrase, a site-specific recombinase, can effectively mediate foreign genes bearing an attB sequence integrated into pseudo attP sites. We have previously identified two pseudo attP sites, BpsF1 and BpsM1 from the bovine genome. In this study, two new pseudo attP sites, BF4 and BF10, were discovered using half-nested inverse PCR from cow fibroblasts. The genomic locations of these two pseudo attP sites were identified by direct sequencing and a BLAST search, and it was confirmed that they reside at positions 4q31 and 10q35 by fluorescence in situ hybridization analysis. Subsequently, the distinct integration frequencies of the four pseudo attP sites were examined. The BF4 site was identified as a hotspot where site-specific integration occurred in most of the cell clones examined, accounting for 74% (42/57) of the integration; much more than the integration frequency for BF10 (7%; 4/57), BpsF1 (7%; 4/57) and BpsM1 (0/57). Interestingly, similar to other hotspots identified in the human and mouse genomes, in which transgenes integrated at hotspots result in high expression, the GFP gene integrated at hotspot BF4 was expressed at high levels in cow fibroblasts, as confirmed by fluorescence microscopy and FACS analysis. Furthermore, ELISA showed that the expression level of the GFP gene integrated at the BF4 site averaged approximately 328 microg x mg(-1), which is more than twofold higher than that integrated at the BF10 site. This study suggests that somatic cells carrying a desired gene integrated at the BF4 site can be used as nuclear donors to generate valuable transgenic animals by nuclear transfer.

[Differential proteomic analysis of human colorectal carcinoma cell lines SW620 and SW480 with different metastatic potentials].

OBJECTIVE: To investigate differential protein expression profiles of human colorectal carcinoma cell lines with different metastatic potentials and screen metastasis-associated proteins for analyzing the relationship between metastasis-associated proteins and the tumorigenesis, progression and metastasis of colorectal carcinomas. METHODS: With two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption/ionization-time of flight-mass spectrometry, we analyzed the differentially expressed proteins in two human colorectal carcinoma cell lines SW620 and SW480 with high and low metastatic potentials, and screened for proteins associated with colorectal carcinoma metastasis. RESULTS: Image analysis software Malenie III demonstrated good match and reproducibility of the 2-DE maps obtained from 3 independent experiments, with 1316+/-62 spots detected for SW620 cells and 1332+/-74 spots for SW480 cells with the average matching rate of 82% and 80%, respectively. The spots distributed in the greatest density at the isoelectric points of 4-7 and relative molecular mass weight of 20,000-70,000. Twenty-five distinctly different protein spots (14 spots for SW620 and 11 for SW480) in-gel digested by TPCK trypsin and 23 peptide mass fingerprint maps were obtained by mass spectrometry. Three highly matched proteins and 14 preliminarily matched proteins or fragments were obtained by analysis with Mascot software in the NCBInr database. Some of these differentially expressed proteins were related to gene transcription, cell cycle, signal transduction, cell apoptosis, etc, with possible involvement of cell differentiation, proliferation, invasion, adhesion, and metastasis of colorectal carcinoma. CONCLUSION: The 2-DE protein expression profile of SW620 cells with high metastatic potential displays obvious difference from that of SW480 cells with low metastatic potential, and a variety of proteins can be involved in the metastasis of colorectal carcinoma.