Histopathological and Immunohistochemical Study of the Distinction between Oral Lichen Planus and Oral Lichenoid Lesions

Background: Oral potentially malignant disorders, which include oral lichen planus (OLP), are clinical presentations that carry a risk of development to cancer in the oral cavity. Oral lichenoid lesions (OLLs) are also termed interface/lichenoid mucositis. Malignant transformation of them remains controversial, but distinct clinical and histological criteria for how to differentiate OLP from OLLs have not been developed. Objectives: The purpose of this study was to elucidate findings that can allow histopathological differentiation of OLP and OLLs using histomorphological and immunohistochemical analyses. Materials and Methods: Analyses were performed in 10 cases diagnosed with OLP and 9 cases diagnosed with OLLs. Cytokeratin 19 (CK19), Ki-67 and CD3 were used as primary antibodies to detect basal cells, proliferative activity and T-cell distribution, respectively, and Perlecan and COX-2 to evaluate epithelial intracellular arrangements and interstitial distributions of proteoglycans and enzymes. Results: For CK19, positive cells were significantly found in OLLs at both the prominent area and site adjacent to the lesion comparison with those of OLP’s. The number of COX-2 positive cells was significantly higher in spinous and basal layers in OLLs of the prominent area. Additionally, OLLs showed mild to moderate expression for perlecan in the basal to spinous layers and in subepithelial tissue. Conclusion: Almost no basal cells were noted in the prominent area in OLP. COX-2 and perlecan were found in the basal to spinous layers in OLLs. Although there are restrictions, these suggested the possibility of helping to distinguish between OLP and OLLs.

Detecting the apoptosis of dopamine neurons with immunohistochemical staining and double-staining technique

BACKGROUND： It is proved that the onset of Parkinson disease companies with neuronal apoptosis of dopamine in substantia nigra of midbrain. Previous researches on neuronal apoptosis of dopamine were analyzed on their consecutive tissue sections with immunohistochemical single-labeling method, immunofluorescence and electron microscope, and there are significant differences.OBJECTIVE ： To observe the feasibility of neuronal apoptosis of dopamine with in situ end labeling and tyrosine-hydroxylase antibody immunohistochemical double-labeling staining technique.DESIGN ： Controlled study.SETTING： College of Pharmacology of Taishan Medical College; College of Management of Taishan Medical College. MATERIALS ： Wistar rats with 2 weeks old and of clean grade were provided by the Animal Center of Taishan Medical College. In situ end labeling kit （terminal deoxynucleotidyl transferase, mixed reactive solution of nucleotide, transfusion-POD）, monoclonal antibody of rat antibody against tyrosine hydroxylase （Boehriuser）. METHODS： The experiment was completed at the Pharmacological Laboratory of Taishan Medical College from February to December 2005. Tissue from midbrain of rats was taken out to make paraffin sections to observe the neuronal apoptosis of dopamine under microscope with in situ end labeling and tyrosine-hydroxylase antibody immunohistochemical double-labeling staining technique.MAIN OUTCOME MEASURES ： Neuronal apoptosis of dopamine with in situ end labeling and tyrosine-hydroxylase antibody immunohistochemical double-labeling staining technique. RESULTS： ① After double-labeling staining, two kinks of positive products were observed in neurons of dopamine which were suffered from apoptosis. One stained with tyrosine hydroxylase was hyacinthine, and the other stained with in situ end labeling was buffy. Cells of positive products stained with in situ end labeling shaped as strap and bend and was distributed in clustering. Cytoplasm was hyacinthine, staining was symmetrical, and cellular ecphyma was observed. Nucleus was stained vacantly which was coincidence with form of neurons of dopamine. ②Apoptosis showed strictly in cytoplasm and nucleus at the aspect of morphology. Cytoplasm stained with in situ end labeling was hardly to recognize because of the usage of double-labeling staining technique, but nucleus was still characterized by apoptosis. The behavior of positive products stained with in situ end labeling was described as following： nucleus was buffy; karyopycnosis was round and irregular; caryotin was integrated into clump which was distributed at the border of nucleus and shaped as demilune and anular; positive signals were limited in nucleus and coincidence with morphological changes of apoptosis. However, blue and positive products were observed in cytoplasm of neurons of dopamine which did not occur apoptosis, and the nucleus was not labeled. Therefore, processing apoptosis of neurons of dopamine could be recognized. CONCULSION： Double-labeling staining technique can be used to correctly reveal histological and morphological changes of neuronal apoptosis of dopamine during its onset and development.

Pancreatic acinar cell carcinoma:A comprehensive review

Acinar cell carcinoma(ACC)is a rare pancreatic malignancy with distinctive clinical,molecular,and morphological features.The long-term survival of ACC patients is substantially superior to that of pancreatic adenocarcinoma patients.As there are no significant patient series about ACCs,our understanding of this illness is mainly based on case reports and limited patient series.Surgical resection is the treatment of choice for patients with the disease restricted to one organ;however,with recent breakthroughs in precision medicine,medicines targeting the one-of-a-kind molecular profile of ACC are on the horizon.There are no standard treatment protocols available for people in which a total surgical resection to cure the condition is not possible.As a result of shared genetic alterations,ACCs are chemosensitive to agents with activity against pancreatic adenocarcinomas and colorectal carcinomas.The role of neoadjuvant or adjuvant chemoradiotherapy has not been established.This article aims to do a comprehensive literature study and present the most recent information on acinar cell cancer.

Is the hypoxia-inducible factor-1 alpha mRNA expression activated by ethanol-induced injury, the mechanism underlying alcoholic liver disease?

BACKGROUND: Excessive alcohol consumption can result in multiple organ injury, of which alcoholic liver disease (ALD) is the most common. With economic development and improvement of living standards, the incidence of diseases caused by alcohol abuse has been increasing in China, although its pathogenesis remains obscure. The aim of this study was to investigate the role of hypoxia in chronic ALD. METHODS: Twenty-eight male Sprague-Dawley rats were randomized into a control group (n=12) with a normal history and an experimental group (n=16) fed with 10 ml/ kg of 56% (vol/vol) ethanol once per day by gastric lavage for 24 weeks. At 24 weeks, blood samples were collected and then the rats were killed. Liver samples were frozen at -80 ℃ and used for RT-PCR; other liver samples were obtained for immunohistochemical staining. RESULTS: When the period of alcohol consumption increased, the positive rate of expression of hypoxia- inducible factor-1 alpha (HIF-1α) mRNA was more significantly elevated in the liver of the alcohol group than in the control group (P≤0.05). The HIF-1α protein located in the cytoplasm was seldom expressed in the control group, but significantly in the alcohol group (P≤0.01). CONCLUSION: HIF-1α mRNA expression was activated by ethanol-induced injury in this study, suggesting that hypoxia is involved in the underlying mechanism of ALD.

The ratio of MMP-2 to TIMP-2 in hilar cholangiocarcinoma:a semi-quantitative study

BACKGROUND: Hilar cholangiocarcinoma is associated with low resectability and poor survival. The aim of this study was to evaluate the roles of matrix metalloproteinases- 2 (MMP-2 ) and its tissue inhibitor of metalloproteinase-2 (TIMP-2) in tumor invasion or as a prognostic factor in patients with human hilar cholangiocarcinoma. METHODS: The expressions of MMP-2 and TIMP-2 were investigated in patients. Paraffinized tissue sections ob- tained from 50 patients with human hilar cholangiocarcino- ma were analysed. The expressions of MMP-2 and TIMP-2 were examined immunohistochemically. Image analysis with image-pro plus analysis software was used to semi- quantitatively determine the ratio of MMP-2 to TIMP-2. RESULTS: The expression levels of MMP-2 and TIMP-2 were strongly associated with tumor hepatic invasion in pa- tients with hilar cholangiocarcinoma. Significant diffe- rences in the ratio of MMP-2 to TIMP-2 between some pathologic factors were observed in patients with hilar cholangiocarcinoma. CONCLUSIONS: MMP-2 plays an essential role in tumor invasion and metastasis,while TIMP-2 is shown to strongly inhibit cancer invasion and metastasis. The ratio of MMP-2 to TIMP-2 may be a prognostic indicator for patients with hilar cholangiocarcinoma.

Breast cancer metastasizing to the stomach mimicking primary gastric cancer: A case report

Breast cancer with stomach metastasis rare with an incidence of 1% or less among metastatic breast cancer patients. We experienced a case of breast cancer metastasizing to the stomach in 65-year-old female patient. She experienced dyspepsia and poor oral intake before visiting the clinic. Diffuse infiltration with nodular mucosal thickening of the stomach wall was observed, suggesting advanced gastric cancer based on gross endoscopic finding. Spread of poorly cohesive tumor cells in the gastric mucosa observed upon hematoxylin and eosin stain resembled signet ring cell carcinoma, but diffuse positive staining for GATA3 in immunohistochemical stain allowed for a conclusive diagnosis of breast cancer metastasizing to the stomach. Based on the final diagnosis, systemic chemotherapy was administered instead of primary surgical resection. After 2 cycles of docetaxel administration, she showed a partial response based on abdominal computed tomography scan. This case is an unusual presentation of breast cancer metastasizing to the gastrointestinal tract.

Detection of Ⅴ,Ⅲ and Ⅰ Type Collagens of Dermal Tissues in Skin Lesions of Patients with Systemic Sclerosis and Its Implication

This study investigated the contents and distribution of collagen Ⅴ （Col Ⅴ） in skin lesions of the patients with systemic sclerosis （SSc） and its roles in the pathogenesis. The contents and distribution for α1 chain of collagen type Ⅰ, Ⅲ and V [α1 （Ⅰ）, α1 （Ⅲ） and α1 （Ⅴ）] in skin lesions of 36 patients with SSc （9 cases of mild fibrosis, 14 moderate, and 13 severe） were detected by using im- munohistochemical SP method. Six cases of normal skin tissues served as controls. The results showed that there was diffuse distribution for three kinds of collagens in dermis. The deep staining α1 （Ⅰ） and α1 （Ⅲ） masses or bands were seen in reticular layer, while α1 （Ⅴ） was distributed more ho- mogeneously. From control to weak, moderate and severe fibrosis stages, α1 （Ⅰ）, α1 （Ⅲ） and α1 （V） showed a gradually increased trend in skin lesions （P〈0.05）. α1 （Ⅴ） was obviously elevated in skin lesions at early stage and persisted in whole fibrotic process and risen in greater contents, while α1 （Ⅰ） and α1 （Ⅲ） were to go higher late and were apparently elevated at moderate and late stages. Com- pared with α1 （Ⅰ）, α1 （Ⅴ） took leading increase at early stage in skin lesions （P〈0.01）, and had more elevated contents than α1 （Ⅲ） at moderate and late stages. The fibrotic changes in dermal reticular layer occurred earlier than those in papillary layer, and the abnormalities of α1 （Ⅴ）/α1 （I） ratio ap- peared before α1 （Ⅲ）/α1 （Ⅰ） ratio. It was concluded that a lot of α1 （Ⅴ） began to deposit in greater contents prior to α1 （Ⅰ） and α1 （Ⅲ） at early stage in SSc and persisted in whole fibrotic process. The changes of α1 （Ⅴ） contents in reticular layer occurred earlier than those in papillary layer, and it sug- gested that the fibrosis in reticular layer appeared earlier.

Early therapeutic effect of platelet-rich fibrin combined with allogeneic bone marrow-derived stem cells on rats’ critical-sized mandibular defects

BACKGROUND Critically sized bone defects represent a significant challenge to orthopaedic surgeons worldwide.These defects generally result from severe trauma or resection of a whole large tumour.Autologous bone grafts are the current gold standard for the reconstruction of such defects.However,due to increased patient morbidity and the need for a second operative site,other lines of treatment should be introduced.To find alternative unconventional therapies to manage such defects,bone tissue engineering using a combination of suitable bioactive factors,cells,and biocompatible scaffolds offers a promising new approach for bone regeneration.AIM To evaluate the healing capacity of platelet-rich fibrin(PRF)membranes seeded with allogeneic mesenchymal bone marrow-derived stem cells(BMSCs)on critically sized mandibular defects in a rat model.METHODS Sixty-three Sprague Dawley rats were subjected to bilateral bone defects of critical size in the mandibles created by a 5-mm diameter trephine bur.Rats were allocated to three equal groups of 21 rats each.Group I bone defects were irrigated with normal saline and designed as negative controls.Defects of group II were grafted with PRF membranes and served as positive controls,while defects of group III were grafted with PRF membranes seeded with allogeneic BMSCs.Seven rats from each group were killed at 1,2 and 4 wk.The mandibles were dissected and prepared for routine haematoxylin and eosin(HE)staining,Masson's trichrome staining and CD68 immunohistochemical staining.RESULTS Four weeks postoperatively,the percentage area of newly formed bone was significantly higher in group III(0.88±0.02)than in groups I(0.02±0.00)and II(0.60±0.02).The amount of granulation tissue formation was lower in group III(0.12±0.02)than in groups I(0.20±0.02)and II(0.40±0.02).The number of inflammatory cells was lower in group III(0.29±0.03)than in groups I(4.82±0.08)and II(3.09±0.07).CONCLUSION Bone regenerative quality of critically sized mandibular bone defects in rats was better promoted by PRF membranes seeded with BMSCs than with PRF membranes alone.

Combined laparoscopic-endoscopic approach for gastric glomus tumor:A case report

BACKGROUND Gastric glomus tumor(GGT)is rare submucosal mesenchymal tumor that lacks specific clinical manifestations and is usually treated mainly by traditional surgical resection.This paper presents a case of a GGT,exhibited both intraluminally and extraluminally growth that was removed by laparoscopy-gastroscopy cooperative surgery.CASE SUMMARY A 52-year-old male presented with epigastric discomfort accompanied by a sense of fullness for 3 mo.Upper gastrointestinal endoscopy identified a submucosal lump located in the gastric antrum.Endoscopic ultrasonography identified a 2.4 cm×1.8 cm lump located in the gastric antrum.It originated from the muscularis propria and exhibited both intraluminally and extraluminally growth,with hypoechoicity on the periphery,hyperechoicity in the middle,and unclear boundaries.Computed tomography showed nodular thickening of 3.0 cm×2.2 cm in the gastric wall of the gastric antrum,and after enhancement,the lesion exhibited obvious enhancement We suspected that it was a gastrointestinal stromal tumor(glomus tumor and schwannoma were not excluded)and planned to perform laparoscopy-gastroscopy cooperative surgery.Immunohistochemical staining after the operation revealed that spinal muscular atrophy(+),hcaldesmon(+),cluster of differentiation 34(CD34)(+),2%Ki-67-positive rate,CD56,melanoma antigen,CD117,discovered on GIST-1,leukocyte common antigen,caudal type homeobox 2,cytokeratin,and S-100 were all negative.The tumor was finally diagnosed as a GGT.CONCLUSION GGTs are rare submucosal tumors of the stomach and should be considered in the differential diagnosis of gastric submucosal tumors.Laparoscopy-gastroscopy cooperative surgery is less invasive and more precise and could be an effective method for the treatment of GGTs.

High expression of squamous cell carcinoma antigen in poorly differentiated adenocarcinoma of the stomach: A case report

BACKGROUND Squamous cell carcinoma antigen(SCCA)is regarded as a specific indicator of epithelial malignancies and is widely used in the diagnosis of squamous cell carcinoma(SCC).However,the expression of SCCA in gastric adenocarcinoma has not been studied in detail.CASE SUMMARY A 52-year-old man was admitted to our hospital for a 2.5 cm x 2.5 cm ulcer at the antrum-body junction with dull pain and fullness in the upper abdomen for 2 mo.His pre-surgery serological testing results showed 0.51 ng/mL SCCA(reference interval,<1.5 ng/mL)and 9.9 ng/mL carcinoembryonic antigen(reference range,<4.7 ng/mL).He underwent radical distal gastrectomy and Roux-en Y anastomosis and was diagnosed with poorly differentiated mucinous adenocarcinoma(Lauren classification:Diffuse)by pathological examination of the resected lesion.Immunohistochemistry showed that SCCA was highly expressed in the cytoplasm of cancer cells.After surgery,the patient received an S-1 adjuvant chemotherapy regimen for six cycles containing tegafur,gimeracil,and oteracil potassium.He showed no sign of recurrence or metastasis within 24-mo follow-up.CONCLUSION This is a frontal report of SCCA overexpression in poorly differentiated adenocarcinoma of the stomach.

Laparoscopic-assisted endoscopic full-thickness resection of a large gastric schwannoma: A case report

BACKGROUND Schwannomas,also known as neurinomas,are benign tumors derived from Schwann cells.Gastrointestinal schwannomas are rare and are most frequently reported in the stomach.They are usually asymptomatic and are difficult to diagnose preoperatively;however,endoscopy and imaging modalities can provide beneficial preliminary diagnostic data.There are various surgical options for management.Here,we present a case of a large gastric schwannoma(GS)managed by combined laparoscopic and endoscopic surgery.CASE SUMMARY A 28-year-old woman presented with a 2-mo history of epigastric discomfort and a feeling of abdominal fullness.On upper gastrointestinal endoscopy and endoscopic ultrasonography,a hypoechogenic submucosal mass was detected in the gastric antrum:It emerged from the muscularis propria and projected intraluminally.Computed tomography showed a nodular lesion(4 cm×3.5 cm),which exhibited uniform enhancement,on the gastric antrum wall.Based on these findings,a preliminary diagnosis of gastrointestinal stromal tumor was established,with schwannoma as a differential.Considering the large tumor size,we planned to perform endoscopic resection and to convert to laparoscopic treatment,if necessary.Eventually,the patient underwent combined laparoscopic and gastroscopic surgery.Immunohistochemically,the resected specimen showed positivity for S-100 and negativity for desmin,DOG-1,α-smooth muscle actin,CD34,CD117,and p53.The Ki-67 index was 3%,and a final diagnosis of GS was established.CONCLUSION Combined laparoscopic and endoscopic surgery is a minimally invasive and effective treatment option for large GSs.

Synchronous Mucinous Borderline Tumor of the Ovary and Low-Grade Appendiceal Mucinous Neoplasm

We present a rare case of synchronous mucinous borderline tumor of the ovary and low-grade appendiceal mucinous neoplasm (LAMN). We performed a left adnexectomy to diagnose the left ovarian borderline malignancy and an ileostomy because of the swollen appendix during the operation. It was diagnosed as left ovarian mucinous borderline malignancy and LAMN. LAMN causes peritoneal dissemination, ovarian metastasis, and peritoneal pseudomyxoma. The appendix and ovary are close to each other anatomically and can metastasize if there is a tumor in either. For ovarian mucinous tumors, it is necessary to search the gastrointestinal tract, especially the appendix, as the primary lesion. For appendix tumors, it is necessary to search for the ovary. Since LAMN may be associated with borderline ovarian malignancies, as in this case, there is a possibility of the duplication of tumors when searching for ovarian mucinous tumors as the primary tumor and if ovarian tumors are found. Since LAMN and mucinous ovarian tumors have similar histological features, immunohistochemical staining is useful for their differentiation because they show different immunostaining patterns.

EXPRESSION AND CLINICAL SIGNIFICANCE OF E2F-3 AND Bc1-2 IN HEPATOCELLULAR CARCINOMA

Objective: To investigate the expression of E2F and Bc1-2 and the clinicopathological significance in hepatocellular carcinoma. Methods: The expressions of E2F-3 and Bc1-2 in 74 patients with hepatic carcinoma, paracarcinoma and 15 patients with liver cirrhosis were detected by S-P immunohistochemical staining. Results: The expression of E2F in hepatic carcinoma was significantly higher than that in paracarcinoma or liver cirrhosis (P<0.005), the expression of Bc1-2 in hepatic carcinoma was significantly higher than that in paracarcinoma (P<0.005), in which Bc1-2 expression was lower than in liver cirrhosis(P<0.05). The expression of E2F-3 was related with histological grade, tumor size, and the expression of Bc1-2 was related with histological grade, tumor size and tumor number. There was correlation between the expression of E2F-3 and Bc1-2 in hepatic carcinoma. Conclusion: E2F-3 and Bc1-2 expression may play an important role in development, progression and cell apoptosis of tumor.

Gastric schwannoma treated by endoscopic full-thickness resection and endoscopic purse-string suture:A case report

BACKGROUND Schwannomas,also known as neurinomas,are tumors that derive from Schwann cells.Gastrointestinal schwannomas are extremely rare,but the stomach is the most common site.Gastric schwannomas are usually asymptomatic.Endoscopy and imaging modalities might offer useful preliminary diagnostic information.However,to diagnose schwannoma,the immunohistochemical positivity for S-100 protein is essential,whereas CD117,CD34,SMA,desmin,and DOG-1 are negative.CASE SUMMARY A 45-year-old female was found to have a gastric mass during a medical examination,which was diagnosed as a gastric schwannoma.We performed endoscopic full-thickness resection and endoscopic purse-string suture.Pathology and immunohistochemical staining confirmed the diagnosis of gastric schwannoma through the positivity of S-100 protein.Furthermore,to exclude the misdiagnosis of gastrointestinal stromal tumor,we performed a mutational detection of the c-Kit and PDGFRA genes.Postoperative follow-up revealed that the patient recovered well.CONCLUSION Immunohistochemical staining is essential for the diagnosis of schwannoma.Endoscopic full-thickness resection is an effective treatment method for gastric schwannoma.

Rare giant corneal keloid presenting 26 years after trauma:A case report

BACKGROUND Corneal keloid is a rare clinical disease with an unknown etiology,which is easily misdiagnosed.Surgery is the most effective treatment but is rarely reported in the literature.Herein,we report the clinical features,histopathology,and surgical outcome of a giant corneal keloid with trophoblastic vessels and discuss the genesis of the mass.CASE SUMMARY A 36-year-old young man was admitted to the hospital because of a large mass on the surface of the left cornea.The patient had suffered an injury to his left eye at the age of 6-years-old;however,as the injury did not cause cornea perforation,he did not undergo treatment.Slit lamp exam showed a large,elevated,opaque lesion that covered the entire cornea and protruded from the surface of the eyeball.Anterior segment optical coherence tomography(AS-OCT) revealed a lesion of irregular density involving the anterior stroma.We suspected a secondary corneal fibroproliferative mass based on the clinical history,and slit lamp and AS-OCT findings.The patient subsequently underwent a superficial keratectomy and keratoplasty,and the final diagnosis of corneal keloid was confirmed by intraoperative histopathological examination.CONCLUSION Non-penetrating corneal trauma damages corneal epithelium basement membrane,initiating stromal fibrosis and causing corneal keloids.AS-OCT and biopsy confirm diagnosis.

Neurothekeoma located in the hallux and axilla:Two case reports

BACKGROUND Neurothekeomas(NTKs)are rare benign soft tissue tumours that typically occur in the head,trunk,and upper limbs and are rare in other parts of the body.CASE SUMMARY Herein,we present two rare cases in which primary NTKs were located in the hallux and axilla.A 47-year-old woman complained of a verrucous bulge on the plantar side of the left hallux.The surface skin of the tumour was abraded due to poor wound healing.A 6-year-old boy complained of a gradually growing subcutaneous mass in the axilla.The tumours of both patients were completely resected,and the diagnosis of NTK was confirmed by histopathology.At the oneyear follow-up,both patients had a good prognosis without local recurrence.CONCLUSION To date,NTKs located in the hallux and axilla have rarely been reported in the literature.We describe NTKs that occurred in unconventional areas and summarize the challenges in their diagnosis and differential diagnosis.

Primary intramedullary melanoma of lumbar spinal cord: A case report

BACKGROUND Primary intramedullary melanoma is a very rare tumor,most frequently occurring in the cervical and thoracic spinal cord.CASE SUMMARY We present a rare case in which the primary intramedullary melanoma was located in the lumbar spine.A 56-year-old man complained of progressive intermittent pain in the lumbar area.Thoracic magnetic resonance imaging showed a spinal intramedullary tumor between the L3 and S1 levels.The tumor was resected entirely,and the diagnosis of malignant melanoma was confirmed by histopathology.CONCLUSION Primary melanoma of the spinal cord,particularly intramedullary localization,has rarely been reported in the previous literature.We describe a primary malignant melanoma of the lumbar spinal cord and discuss the challenges associated with the diagnosis.

Pancreatic cancer with synchronous liver and colon metastases: A case report

BACKGROUND Metastasis of pancreatic cancer to the colon is rare and the features need to be further elucidated.Herein,we report a rare case of pancreatic cancer with simultaneous liver and colon metastases.CASE SUMMARY A 48-year-old man with intrahepatic space-occupying lesions based on a computed tomography scan was admitted to our hospital for further treatment.Abdominal magnetic resonance imaging revealed a 6.4 cm×4.2 cm mass in the tail of the pancreas and multiple low-density masses in the liver parenchyma.In addition,a mass of 2.2 cm×1.6 cm with surface congestive erosions in the sigmoid colon was detected by colonoscopy.Histopathological examination of biopsies from both the liver and colon lesions revealed a moderately to poorly differentiated adenocarcinoma.Immunohistochemical staining of the colon tumor was positive for cytokeratin(CK)7 and CK,but negative for colorectal adenocarcinoma-related markers CK 20,CDX2,and SATB2,thus indicating that the metastasis originated from the pancreas.Next-generation sequencing for genomic profiling of the liver and colon metastases both found mutations in KRAS(p.G12D)and TP53(c.376-1delG),with microsatellite stable and low tumor mutational burden without actionable or cancer-predisposing gene mutations detected.The patient was subsequently treated with 12 cycles of FOLFIRINOX which led to a sustainable response,followed by ongoing maintenance treatment with irinotecan plus fluorouracil.CONCLUSION For this rare case,careful evaluation of histopathological and immunohistochemical staining results are required.The genomic profiling of colon lesions was revealed for the first time,and FOLFIRINOX showed good treatment efficacy in this patient.

A Statistical Evaluation of Uncoupling Protein 1 in the Limited Area of Brown Adipose Tissue by Immunoelectron Microscopy

Uncoupling protein 1 (UCP1) expressed by the brown adipose tissue (BAT) in the mitochondrial crista acts as a homeostatic thermogenerator of eutherians. The evaluation of UCP1 expression in the BAT offers significant scientific insight, especially in studies targeting limited areas such as the periarterial and pericardial regions of small experimental mammals. However, the negligible amount of this adipose tissue would render the general quantitative evaluation of the protein unreliable because of lipid contamination and low protein concentration. To address this problem, we quantitatively evaluated UCP1 expression in the mitochondrion of the mouse interscapular BAT using immunoelectron microscopy and immunohistochemical studies using a combination of primary and secondary antibodies in scheme A (rabbit anti-UCP1 IgG/gold particle-conjugated goat anti-rabbit IgG), B (rabbit IgG/gold particle-conjugated goat anti-rabbit IgG), C (rabbit anti-UCP1 IgG/gold particle-unconjugated goat anti-rabbit IgG), and D (rabbit IgG/gold particle-unconjugated goat anti-rabbit IgG). Scheme A shows the immunopositive reaction of obvious gold particles in the mitochondrial area, whereas other procedures revealed less distinctive reactions. The distinctive gold particle immunoreaction comprised electrical high-density spots with a mean diameter of >5 nm. However, in scheme B, the electrical high-density spots were scattered outside the mitochondrion and were significantly smaller than 4 nm;schemes C and D demonstrated few immunoreactions. Logistic regression analysis between schemes A and B showed that the threshold diameter of the electrical high-density spots measuring >5 nm indicated a true positive immunoreaction to anti-UCP1 antibody specifically in the mitochondrial area. Minor statistical difference was observed in the primary anti-UCP1 antibody between polyclonal IgG and monoclonal antibodies. Therefore, immunoelectron microscopy might be useful for evaluating negligible protein expression in some limited areas, such as UCP1 expression in the BAT of small experimental animals.

Tissue distribution of deoxynivalenol in piglets following intravenous administration

Contamination of deoxynivalenol（DON） in grains is common worldwide and pigs are particularly susceptible to this mycotoxin. The distribution of DON in porcine tissues following intravenous administration was investigated in this study. Fifteen pigs were randomly divided into three groups. Animals in groups A and B were administrated with DON at the dose of 250 and 750 μg kg–1 body weight, respectively, while group C served as blank control. Plasma, bile and 27 tissues were collected at 30 min post-administration. DON concentrations in all samples were tested using high-performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS）. To observe the distribution of DON in tissues, these samples were further subjected to the immunohistochemical analyses. Totally, the bile and 13 tissues were sampled for DON-based detection, including kidney, mesenteric lymph nodes, muscle, stomach, jejunum, colon, plasma, spleen, rectum, cecum, liver, ileum, and duodenum. No significant difference was observed for the concentrations of DON in duodenum, ileum and liver samples between groups A and B; while the DON concentrations in cecum and rectum of group B were significantly higher（P-value 〈0.05） than those in group A. In addition, the DON concentrations in stomach, jejunum, colon, mesenteric lymph nodes, muscle, kidney, spleen, bile, and plasma of group B were remarkably higher than those of group A（P-value〈0.01）. Levels of DON in other 14 tissues including medulla oblongata, midbrain, diencephalon, pons, tip and tongue body, tongue, soft palate, tonsils, pharyngeal mucosa, oral buccal mucosa, thymus, thyroid, esophagus and adrenal gland were all below the limit of detection. The results of immunohistochemistry showed that 11 tissue samples（medullaoblongata, tonsil, adrenal medulla, thyroid gland, thyroid, stomach, duodenum, jejunum, kidney, spleen, and mesenteric lymph nodes） were positive and DON was mainly distributed around blood vessels in these tissues. Therefore, we believed that concentrations of DON in tissues differ when pigs are in exposure to various dosages and DON causes lesions in many pig tissues.