Expression of heat shock proteins (HSP27, HSP60, HSP70, HSP90, GRP78, GRP94) in hepatitis B virus-related hepatocellular carcinomas and dysplastic nodules

AIM: Expression of heat shock proteins (HSPs) is frequently up-regulated in hepatocellular carcinoma (HCC), which evolves from dysplastic nodule (DN) and early HCC to advanced HCC. However, little is known about the differential expression of HSPs in multistep hepatocarcinogenesis. It was the purpose of this study to monitor the expression of HSPs in multistep hepatocarcinogenesis and to evaluate their prognostic significance in hepatitis B virus (HBV)related HCC.METHODS: Thirty-eight HCC and 19 DN samples were obtained from 52 hepatitis B surface antigen-positive Korean patients. Immunohistochemical and dot immunoblot analyses of HSP27, HSP60, HSP70, HSP90, glucoseregulated protein (GRP)78, and GRP94 were performed and their expression at different stages of HCC development was statistically analyzed.RESULTS: Expression of HSP27, HSP70, HSP90, GRP78, and GRP94 increased along with the stepwise progression of hepatocarcinogenesis. Strong correlation was found only in GRP78 (Spearman's r= 0.802). There was a positive correlation between the expressions of GRP78, GRP94, HSP90, or HSP70 and prognostic factors of HCC. Specifically, the expression of GRP78, GRP94, or HSP90 was associated significantly with vascular invasion and intrahepatic metastasis.CONCLUSION: The expressions of HSPs are commonly up-regulated in HBV-related HCCs and GRP78 might play an important role in the stepwise progression of HBVrelated hepatocarcinogenesis. GRP78, GRP94, and HSP90 may be important prognostic markers of HBV-related HCC, strongly suggesting vascular invasion and intrahepatic metastasis.

Nogo-A expression dynamically varies after spinal cord injury

The mechanism involved in neural regeneration after spinal cord injury is unclear. The my-elin-derived protein Nogo-A, which is speciifc to the central nervous system, has been identiifed to negatively affect the cytoskeleton and growth program of axotomized neurons. Studies have shown that Nogo-A exerts immediate and chronic inhibitory effects on neurite outgrowth.In vivo, inhibitors of Nogo-A have been shown to lead to a marked enhancement of regenerative axon extension. We established a spinal cord injury model in rats using a free-falling weight drop device to subsequently investigate Nogo-A expression. Nogo-A mRNA and protein expression and immunoreactivity were detected in spinal cord tissue using real-time quantitative PCR, immu-nohistochemistry and western blot analysis. At 24 hours after spinal cord injury, Nogo-A protein and mRNA expression was low in the injured group compared with control and sham-operated groups. The levels then continued to drop further and were at their lowest at 3 days, rapidly rose to a peak after 7 days, and then gradually declined again after 14 days. These changes were observed at both the mRNA and protein level. The transient decrease observed early after injury followed by high levels for a few days indicates Nogo-A expression is time dependent. This may contribute to the lack of regeneration in the central nervous system after spinal cord injury. The dynamic varia-tion of Nogo-A should be taken into account in the treatment of spinal cord injury.

Differential expression of Rab27A/B correlates with clinical outcome in hepatocellular carcinoma

AIM:To investigate the association of Rab27A and Rab27B expression with clinicopathological characteristics and prognosis of hepatocellular carcinoma(HCC).METHODS:We used reverse transcription polymerase chain reaction(RT-PCR),real-time PCR,and Western blotting to detect Rab27A and Rab27B mRNA and protein expression in 5 human HCC lines and the immortalized hepatic HL-7702 cell line.We further examined 148 primary HCC samples matched with adjacent normal tissue and 80 non-HCC specimens by immunohistochemistry to evaluate the correlation of Rab27A and Rab27B expression with clinicopathological features and prognosis.RESULTS:Our data showed that Rab27A and Rab27Bwere differentially expressed in cell lines and primary HCC tumors.Rab27A mRNA and protein were detected in 67%(4/6)of human cell lines and 80%(4/5)of HCC cell lines,while Rab27B was found in 50%(3/6)of human lines and 40%(2/5)of HCC lines.Rab27A expression was higher in primary HCC(46.2%,66/143)than in matched adjacent tissue(24.3%,33/136,P<0.001),whereas immunopositivity for Rab27B was lower in primary HCC(57.4%,81/141)than in matched adjacent tissue(87.5%,119/136,P<0.001).Analysis of clinicopathological characteristics of 148 HCC specimens revealed significant correlations between Rab27A and Rab27B expression and tumor tumor-node-metastasis(TNM)classification(P=0.046 and P=0.027,respectively),and between strong Rab27A expression and tumor differentiation grade(P=0.008).Survival analyses revealed that patients with Rab27A+or Rab27B+tumors had significantly reduced overall survival compared with that of patients with Rab27A-or Rab27B-tumors(P= 0.015 and P=0.005,respectively).Risk analyses revealed that Rab27B+and TNMⅢ-Ⅳwere independent poor prognosis factors associated with a 3.36-and 3.37fold higher relative risk of death,respectively.CONCLUSION:Rab27A and Rab27B expression were closely correlated with tumor progression and can be valuable prognostic indicators for HCC patients.

Endobronchial metastasis from adenocarcinoma of gastric cardia 7 years after potentially curable resection

Endobronchial metastasis(EBM) is a rare form of metas-tasis from extrapulmonary malignant tumors,although there are few reports of EBM from gastric cancer specifically.We report the case of a 51-year-old woman who had undergone gastrectomy for advanced gastric cancer seven years previously but was diagnosed with a solitary lung tumor by follow-up computed tomography.On diagnosis of primary lung cancer,she underwent pulmonary lobectomy,but immunohistochemical examination confirmed the resected tumor to be an EBM from the gastric cancer.Six months later,she was diagnosed with peritoneal metastases and underwent chemotherapy with gastric cancer regimen.She is still alive at 33 mo after the lobectomy.Generally,the prognosis for EBM is poor although multidisciplinary treatment can lead to long-term survival.Precise diagnosis on the basis of detailed pathological and immunohistochemical evaluation can contribute to deciding the most effective treatment and improving prognosis.

Changes of 5-hydroxytryptamine and tryptophan hydroxylase expression in the ventral horn of spinal cord

Objective To investigate changes of 5-hydroxytryptamine （5-HT） and its synthesis rate-limiting enzyme tryp-tophan hydroxylase （TPH） in the ventral horn of spinal cord after exercise-induced fatigue, and to further discuss the mecha- nism of exercise-induced central fatigue at spinal level. Methods Sixteen healthy adult Wistar rats were randomly divided into 2 groups： exercise-induced fatigue group and control group. Immunohistochemical staining for 5-HT and TPH in the ventral horn were performed and analysized quantitatively. The mean optic densities of 5-HT and TPH positive fibers or terminals were measured by computerized image analyzer. Results Both 5-HT and TPH positive fibers/terminals decreased in the exercise-induced fatigue group. The immunohistochemical staining was weaker and the mean optic densities decreased obviously in the fatigue group compared with those in the control group （P 〈 0.05）. Conclusion 5-HT and TPH in the ventral horn of spinal cord might be involved in exercise-induced fatigue.

Chromophobe renal cell carcinoma: Novel molecular insights and clinicopathologic updates

Chromophobe renal cell carcinoma(ChRCC)is the third most common renal cell carcinoma(RCC)subtype,which predominantly occurs in sporadic setting.ChRCCs are considered to originate from the intercalated cell of distal tubules with two main morphological variants,classic and eosinophilic.Most ChRCCs carry a favorable clinical outcome.Histology alone is limited in predicting the behavior of ChRCCs that do not have overtly aggressive morphologic findings such as necrosis and sarcomatoid features.Along with positive CD117 expression,classic ChRCCs generally express diffuse and uniform CK7,while eosinophilic variant demonstrates more heterogeneous CK7 expression(rare or patchy).Multiple losses of chromosomes 1,2,6,10,13,17,and 21 are considered to be the genetic hallmarks of classic and eosinophilic ChRCCs,while chromosomal gains are known to be associated with sarcomatoid ChRCCs.TP53 and PTEN are the two most frequently mutated genes in ChRCCs.The major challenge in the differential diagnosis of ChRCCs includes considerations around the eosinophilic variant(of ChRCCs),where it may share overlapping features with oncocytoma or other recent emergent oncocytic tumors.Most eosinophilic ChRCCs share expression of the recently described biomarkers,LINC01187 and FOXI1,with classic ChRCCs,however,a subset of eosinophilic-like ChRCCs with lower biomarker expression have been demonstrated to harbor MTOR gene mutations.Overall,the morphologic features of ChRCCs and genetic profile with combinations of chromosomal losses and gains suggest this tumor entity to represent a distinct,yet heterogeneous group of renal neoplasms.

EXPRESSION OF EPIDERMAL GROWTH FACTOR, TRANSFORMINGGROWTH FACTOR-α AND THEIR RECEPTOR IN HUMAN PITUITARY TUMORS

Objective: To explore the role of growth factor autocrine stimulation in the pathogenesis of human pituitary tumors. Methods: The expression of EGF, TGF-( and EGFR were studied by immunohisto-chemical method on paraffin-embedded sections of 30 cases pituitary tumor. Results: EGFR and its ligands EGF, TGF-( expressed in majority of pituitary tumors. The expression of EGFR and its ligands varied with cells' intensity, density and type. Conclusion: The EGF autocrine stimulating exerted in the pituitary tumor development process, that tyrosine kinases inhibitors may be useful for pituitary tumors treatment.

Pathological and immunohistochemical findings of lungs,heart,liver,and kidneys,and unexpected findings of fungi and parasites in lungs of deceased COVID-19 patients:A case series

Objective:To define histopathologic and immunohistochemical features of the lungs,heart,liver,and kidneys in patients who died from coronavirus disease 2019(COVID-19),and to determine the presence of SARS-CoV-2 in all tissues,as well as the presence of fungi and parasites in lung tissues.Methods:This retrospective case study was conducted in the intensive care units of Dokuz Eylül University Hospital,and patients(≥18 years)who died due to COVID-19 between October 2020 and April 2021 were included.The biopsy samples of the patient’s lung,heart,liver,and kidney tissues were studied.Results:In the study,we enrolled 12 patients(mean age:70 years;50%male).Alveolar epithelial cell damage and diffuse alveolar damage were predominant in lung tissues.Lobular lymphocyte infiltration,centrilobular sinusoidal dilatation,and microvesicular steatosis in the liver,together with pigmented cast,non-isometric vacuolar degeneration,and capillary plugging in the kidneys,were commonly found among the patients.SARS-CoV-2 nucleocapsid protein antibodies were detected in three lung and two kidney tissues,and so did angiotensin-converting enzyme 2 receptor positivity in one lung and more than half of the kidney tissues.The RT-PCR tests were positive in three lungs and one kidney tissue.After DNA isolation from lung tissues,Pneumocystis jirovecii was detected in nine patients,Aspergillus fumigatus in two,Microsporidia in three,and Cryptosporidium in two.Conclusions:SARS-CoV-2 is a multisystemic disease.Fungi and parasites should be investigated in critically ill COVID-19 patients prescribed corticosteroids.

Kaposiform Hemangioendothelioma:a Case Report

Case Report A 36-year-old man with a neoplasm in his right-leg had undergone surgical removal of this neoplasm at an out-patient clinic (OPD). The tumor size was about 1 cm×1 cm×0.5 cm. It was knotty and nonencapsulated. Its cross-section was kermesinus and its texture was moderate. The pathological examination confirmed a diagnosis of Kaposiform hemangioendothelioma.

Morphological properties of nociceptive and non-nociceptive neurons in primary somatic cerebral cortex (SI) of cat

With the techniques of intracellular recording and labelling, we investigated pain sensation and modulation of the somatic cortical cortex at the neuron’s level. After observing the evoked potentials from stimulating the saphenous nerves (SN) of 654 neurons in SI area of the cats, we labelled 30 of the neurons with Neurobiotin to preserve the distribution and the morphologic characteristics of the neurons in the cortex. Based on the tridimensional reconstruction in addition to the eletrophysiological functions, we found clear morphological distinctions between nociceptive and non-nociceptive neurons (P【0.01). This result provided new experimental material to illustrate the function of nociceptive neurons in somatosensory cortex (SI) and presented further evidence to support the 'specificity theory' of pain sensation in terms of morphology.