Analysis of intraoperative frozen section diagnosis of 1 263 pulmonary nodules / 中国胸心血管外科临床杂志

@#Objective    To explore the key points and difficulties of intraoperative frozen section diagnosis of pulmonary diseases. Methods    The intraoperative frozen section and postoperative paraffin section results of pulmonary nodule patients in Beijing Chaoyang Hospital, Capital Medical University from January 2021 to January 2022 were collected. The main causes of misdiagnosis in frozen section diagnosis were analyzed, and the main points of diagnosis and differential diagnosis were summarized. Results    According to the inclusion criteria, a total of 1 263 frozen section diagnosis results of 1 178 patients were included in the study, including 475 males and 703 females, with an average age of 58.7 (23-86) years. In 1 263 frozen section diagnosis results, the correct diagnosis rate was 95.65%, and the misdiagnosis rate was 4.35%. There were 55 misdiagnoses, including 18 (3.44%) invasive adenocarcinoma, 17 (5.82%) adenocarcinoma in situ, 7 (35.00%) mucinous adenocarcinoma, 4 (2.09%) minimally invasive adenocarcinoma, 3 (100.00%) IgG4 related diseases, 2 (66.67%) mucinous adenocarcinoma in situ, 1 (16.67%) atypical adenomatous hyperplasia, 1 (14.29%) sclerosing pulmonary cell tumor, 1 (33.33%) bronchiolar adenoma, and 1 (100.00%) papillary adenoma. Conclusion    Intraoperative frozen section diagnosis still has its limitations. Clinicians need to make a comprehensive judgment based on imaging examination and clinical experience.

Survivin expression as a prognostic marker for breast cancer

SUMMARY OBJECTIVE: Due to the speed of development observed in breast cancer, several studies aimed at discovering new biomarkers have been carried out in order to arrive at an early diagnosis. As survivin plays a fundamental role in the evasion of apoptosis in tumor cells, the aim of this study was to verify the expression profile of the survivin gene in paraffin-embedded breast tumor samples and associate it with the clinical characteristics of the patients. METHODS: This is a cross-sectional study, for which 100 tumor samples were obtained from cancer patients treated throughout the year 2019 at Instituto de Mama do Cariri (Juazeiro do Norte, in the state of Ceará). This study included women over 30 years old who had confirmed breast cancer through anatomopathological examination but excluded those with non-neoplastic breast comorbidities, other neoplasms, or chronic diseases. Survivin gene expression was assessed by quantitative polymerase chain reaction. RESULTS: The expression of survivin is associated with the lack of expression of estrogen (p=0.027) and progesterone (p>0.0005) receptors. It means that survivin expression is higher in patients in which labeling was absent for estrogen receptor and progesterone receptor. CONCLUSION: Our data reinforce that survivin expression is higher in estrogen receptor-patients, thus representing an additional prognostic tool.

Utility of serum anti-phospholipase A2 receptor antibody as a biomarker in membranous nephropathy: An experience from a tertiary care center of Eastern India

Background: Membranous nephropathy (MN) is a pattern of glomerular injury. Exact categorization into primary membranous nephropathy (PMN) or secondary membranous nephropathy (SMN) is essential for treatment. An endogenous podocyte antigen, M-type phospholipase A2 receptor (PLA2R) has been discovered to be involved in the pathogenesis of PMN. Aims and Objectives: In this article, we aimed to analyze renal tissue PLA2R and serum anti-PLA2R antibodies in MN cases and determined the diagnostic utility. Materials and Methods: The study was of prospective type carried out from March 2019 to August 2020. Analysis of cases of MN was performed with PLA2R paraffin immunoflourescence and serum anti-PLA2R antibody ELISA. Results: Overall sensitivity, specificity, PPV, and NPV of serum anti-PLA2R ELISA for PMN was 91.3%, 80%, 75%, and 93.3%, respectively, and of tissue PLA2R staining for PMN was 91.67%, 81.08%, 75.86%, and 93.75%, respectively. There was strong concordance between two methods. In the patients that were followed up, we found baseline serum anti-PLA2R antibody was less in complete remission group than that in non-remission group and the reduction in serum anti-PLA2R antibody was more in complete remission group than that in non-remission group. Conclusion: Routine light and immunofluorescence examination are incapable of giving exact categorical opinion regarding PMN and SMN. Serum anti-PLA2R antibody detection and renal tissue PLA2R analysis are sensitive and specific in detecting PMN. Baseline serum anti-PLA2R antibody and anti-PLA2R antibody quantification trends are related to prognosis of PMN. So they can be incorporated as additional biomarker.

Value of cell paraffin block immunohistochemistry and pleural effusion CRKL and MIC-1 in the diagnosis of malignant pleural effusion / 国际肿瘤学杂志

Objective:To study the value of cell paraffin block immunohistochemistry and pleural fluid Crk like protein (CRKL) and macrophage inhibitory cytokine-1 (MIC-1) in the diagnosis of malignant pleural effusion.Methods:A total of 98 patients with pleural effusion treated in Shantou Central Hospital from February 2020 to February 2021 were retrospectively selected as the research objects, including 58 benign cases and 40 malignant cases. The levels of CRKL and MIC-1 in pleural effusion were detected by enzyme-linked immunosorbent assay. The pleural effusion was analyzed by cell paraffin block immunohistochemistry. The levels of various indexes in benign group and malignant group were compared. The diagnostic value of cell paraffin block immunohistochemistry and pleural effusion CRKL and MIC-1 for benign and malignant pleural effusion was analyzed by receiver operating characteristic (ROC) curve.Results:With pathological results as the gold standard, 54 cases of benign and 44 cases of malignant were diagnosed by cell paraffin block immunohistochemistry. The diagnostic accuracy was 75.5% (74/98) , and the sensitivity and specificity were 75.0% (30/40) and 75.9% (44/58) respectively. The levels of pleural effusion CRKL [2.84 (2.17, 3.98) ng/ml vs. 1.88 (0.94, 2.62) ng/ml], MIC-1 [2.28 (1.67, 2.98) ng/ml vs. 1.76 (1.22, 2.32) ] ng/ml] in the malignant group were higher than those in the benign group, with statistically significant differences ( Z=-4.57, P<0.001; Z=-3.09, P<0.001) . The optimal critical value of CRKL in pleural effusion for the diagnosis of malignant pleural effusion was 2.33 ng/ml, the area under the curve (AUC) was 0.76 (95% CI: 0.66-0.85) , and the sensitivity and specificity were 67.5% (27 /40) , 74.1% (43/58) . The optimal critical value of MIC-1 in pleural effusion for the diagnosis of malignant pleural effusion was 2.10 ng/ml, the AUC was 0.74 (95% CI: 0.64-0.85) , and the sensitivity and specificity were 60.0% (24/40) , 82.8% (48/58) . The AUC of MIC-1 and CRKL in pleural effusion combined with cell paraffin block immunohistochemistry for the diagnosis of malignant pleural effusion was 0.83 (95% CI: 0.75-0.91) , and the sensitivity and specificity were 85.0% (34/40) and 70.7% (41/58) . The sensitivity and AUC of combined diagnosis were significantly higher than those of CRKL and MIC-1 alone (sensitivity: χ2=4.26, P=0.046; χ2=6.27, P=0.012; AUC: Z=3.53, P<0.001; Z=4.14, P<0.001) . Conclusion:CRKL and MIC-1 in pleural effusion of patients with malignant pleural effusion are highly expressed, which can be used as indicators for the diagnosis of malignant pleural effusion. Detection combined with cell paraffin block immunohistochemistry can improve the diagnostic value of malignant pleural effusion.

Experimental study on the structural and functional changes of the retinal tissue induced by intense blue light irradiation in rats / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To observe the structural and functional changes of retinal tissue in rats after different duration of intense blue light irradiation.METHODS: A total of 48 healthy 8-week-old SD male rats were selected and randomly divided into the control group(n=12)and 3, 6 and 12h experimental groups(n=36). The rats in the control group received natural light, and the rats in the experimental groups received blue light with a wavelength of 465±5nm and the illumination of 1000±100lx for 3, 6, and 12h each day. Optical coherence tomography(OCT), fundus fluorescein angiography(FFA)and haematoxylin-eosin(HE)staining of paraffin pathological section were used to observe the changes of the retinal thickness, retinal tissue structure and the function in different directions and layers.RESULTS: The OCT results showed that the retinal thickness in the superior, inferior, nasal, and temporal sides of rats in different groups was statistically significant(P&#x0026;#x003C;0.05), and there was no statistical significant difference between the control group and the 3h experimental group in the total retinal thickness(P&#x0026;#x003E;0.05), while the differences between any other two groups were statistically significant(P&#x0026;#x003C;0.05); The mean total retinal thickness, internal limiting membrane(ILM)-inner nuclear layer(INL)thickness, outer plexiform layer(OPL)-outer segment(OS)thickness and retinal pigment epithelium(RPE)of rats in each group were statistically significant(P&#x0026;#x003C;0.05), and the mean total retinal thickness and OPL-OS thickness were statistically significant between any two groups(P&#x0026;#x003C;0.05). The ILM-INL thickness of rats in the control group and 3 and 12h experimental groups was statistically significant(P&#x0026;#x003C;0.05), and the thickness of RPE layer in the 12h experimental group was statistically different from that of the 3 and 6h experimental groups(P&#x0026;#x003C;0.05). FFA results showed that there was no obvious fluorescence leakage in the fundus of rats in the control group and the 3h experimental group, while there was obvious fluorescence leakage and hyperfluorescence in the retina of the 6 and 12h experimental groups, and the background fluorescence of choroid was enhanced. HE staining showed the atrophy and apoptosis of cells in the optic cell layer, and some lightly stained nucleus. In addition, RPE layer showed atrophy and thinned with the increase of light time, and there was significant difference in the number of the optic cells between the control group and experimental group(P&#x0026;#x003C;0.05).CONCLUSION: The intense blue light irradiation could cause thinning of the retina in rats, with varying degrees of thinning in different layers of the retina. It could also led to decrease and even disappearance of the number of cells in the visual cell layer, the focal atrophy of the RPE layer, and the change of vascular permeability. With the extension of the light time, the structural and functional changes in retinal tissue became more obvious.

Intraoperative assessment of surgical margins using ‟en face" frozen sections in the management of cutaneous carcinomas

Abstract Background: Basal cell and squamous cell carcinomas (BCC and SCC) are the most common types of cancer worldwide. Intraoperative assessment of surgical margins by frozen section has been widely used to ensure disease-free margins. The intraoperative ‟en face" freezing technique evaluates all peripheral and deep margins. Objective: To report the results of the ‟en face" freezing technique in relation to tumor recurrence and agreement with paraffin-embedded tissue examination. Methods: Retrospective analysis of patients undergoing surgical excision of BCC and SCC at the A. C. Camargo Cancer Center, Brazil. Results: This study included 542 skin carcinomas, which were excised from 397 patients. A total of 201 male patients (50.6%), and 196 female patients (49.4%) were assessed, whose mean age was 64 years. The tumors were mostly located on the head and neck region (87.8%). BCC corresponded to 79.7% of the cases. The mean follow-up was 38 months. Tumor relapse occurred in 0.86% of the primary tumors and 3.7% of recurrent tumors. The result of the intraoperative ‟en face" frozen section evaluation was in agreement with the final result of the anatomopathological examination (paraffin test) in 98% of the lesions. Study limitations: Not having a minimum follow-up time of 5 years for all patients. Conclusion: The ‟en face" freezing technique shows low tumor relapse, being reliable and safe to guarantee negative surgical margins of the tumor.

Cell HE staining smears and paired cell paraffin sections in detection of epithelial growth factor receptor gene of pleural fluid specimens / 中南大学学报(医学版)

OBJECTIVES@#The advanced non-small cell lung cancer (NSCLC) patients with pleural effusion have no opportunity for surgery treatment. Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are the first-line drugs for these patients with EGFR-sensitive mutation. However, the disease progression and drug update during or after treatment of EGFR-TKIs bring more challenges and puzzles to clinical diagnosis and treatment, which inevitably requires archived pleural cell samples for EGFR re-examination or comparative study. Understanding the DNA quality of archived pleural fluid samples and effectively using archival data of pleural fluid cells are of great significance for tracing the origin of cases and basic medical research. This study aims to evaluate the consistency of EGFR mutant gene expression between the 2 methods, and to explore a reliable way for preserving cytological data and making full use of cytological archival data via cell HE staining smear and cell paraffin section.@*METHODS@#A total of 57 pleural fluid cytology cases in the Department of Pathology of China Aerospace Center Hospital from October 2014 to April 2021 were selected. Tumor cells were detected by cell HE staining smears and immunohistochemical staining for TTF-1 and Napsin A in the paired cell paraffin sections. There were more than 200 tumor cells in cell HE staining smear and the proportion of tumor cells were ≥70% in matched cell paraffin sections. Patients with 2 cell smears (one for cell data retention and the other for DNA extraction) were selected as the research subjects, and 57 pleural fluid samples were enrolled. EGFR gene mutation was detected by amplification refractory mutation system-polymerase chain reaction in 57 paired cell HE staining smears and cell paraffin sections. DNA concentration was 2 ng/μL. Cell HE smear was amplified side-by-side with DNA samples from paired cell paraffin sections. Result determination was according to the requirements of the reagent instructions. The external control cycle threshold (Ct) value of the No. 8 well of the samples to be tested was between 13 and 21, which was considered as successful and reliable samples. When the Ct value of EGFR gene mutation was <26, it was considered as positive; when the Ct value was between 26 and 29, it was critical positive; when the Ct value was equal or more than 29, it was negative. ΔCt value was the difference between mutant Ct value and externally controlled Ct value. The smaller the ΔCt value was, the better the quality of DNA of the detected sample was.@*RESULTS@#Among the 57 pleural effusion samples, 42 patients were hospitalized with pleural effusion as the first symptom, accounting for 73.7% (42/57). EGFR mutation was detected in 37 samples [64.9% (37/57)]. The mutation rate for 19del was 37.8% (14/37) while for L858R was 48.6% (18/37). Females were 56.7% (21/37) of mutation cases. The mutation consistency rate of cell HE staining smear and matched cell paraffin sections was 100%. The ΔCt values of cell HE staining smears were less than those of matched cell paraffin sections. The mutation Ct values of 37 cytological samples were statistically analyzed according to the preservation periods of the years of 2014-2015, 2016-2017, 2018-2019, and 2020-2021. There were significant differences in cell paraffin section in the years of 2014-2015 and 2016-2017 compared with the years of 2018-2019 and 2020-2021, while no significant differences were found in cell HE staining smear. Statistical analysis of externally controlled Ct values of 57 cytological samples showed that there were significant differences between cell HE staining smears and cell paraffin section in the years of 2014-2015 and 2016-2017, compared with the years of 2018-2019 and 2020-2021. The mutational Ct values of 37 paired cell blocks and smears were all <26, and the externally controlled Ct values of 57 paired cell paraffin sections and HE staining smears were all between 13 and 21.@*CONCLUSIONS@#The DNA quality of cell HE smears and matched cell paraffin section met the qualified requirements. Two methods possess show an excellent consistency in detecting EGFR mutation in NSCLC pleural fluid samples. The DNA quality of cell HE staining smear is better than that of cell paraffin sections, so cell HE staining smear can be used as important supplement of the gene test source. It should be noted that the limitation of cell HE staining smears is non-reproducibility, so multiple smears of pleural fluid are recommended to be prepared for multiple tests.

Practical tips to using formalin-fixed paraffin-embedded tissue archives for molecular diagnostics in a South African setting

Background: Formalin-fixed paraffin-embedded (FFPE) tissue archives in hospitals, biobanks, and others offer a vast collection of extensive, readily available specimens for molecular testing. Unfortunately, the use of tissue samples for molecular diagnostic applications is challenging; thus, the forensic pathology FFPE tissue archives in Africa have been a largely unexploited genetic resource, with the usability of DNA obtainable from these samples being unknown.Intervention: The study, conducted from January 2015 to August 2016, determined the usefulness of FFPE tissue as a reliable source of genetic material for successful post-mortem molecular applications and diagnostics. Formalin-fixed paraffin-embedded tissue samples were collected and archived from autopsies conducted over 13 years in the forensic medicine department of the University of Pretoria (Pretoria, South Africa). Deoxyribonucleic acid from FFPE tissue samples and control blood samples was amplified by high-resolution melt real-time polymerase chain reaction before sequencing. The procurement parameters and fixation times were compared with the quantity and quality of the extracted DNA and the efficiency of its subsequent molecular applications.Lessons learnt: This study has shown that FFPE samples are still usable in molecular forensics, despite inadequate sample preparation, and offer immense value to forensic molecular diagnostics.Recommendations: FFPE samples fixed in formalin for more than 24 h should still be used in molecular diagnostics or research, as long as the primer design targets amplicons not exceeding 300 base pairs.

Evaluación de la metilación del DNA en tejidos de cáncer colorrectal fijados en formalina y embebidos en parafina (FFPE) / Evaluation of DNA methylation in formalin-fixed, paraffin embedded colorectal cancer tissues (FFPE)

Resumen Las alteraciones en la metilación de dinucleótidos CpG en regiones promotoras es uno de los mecanismos epigenéticos implicados en cáncer que tiene uso potencial como biomarcador. Su evaluación, a partir de tejidos fijados en formalina y embebidos en parafina (FFPE), representa un gran desafío dadas la degradación parcial, el entrecruzamiento y las bajas cantidades del DNA obtenido. En esta nota técnica, describimos un protocolo para el estudio del estado de metilación del promotor distal del proto-oncogén K-RAS, a partir de varias muestras obtenidas de dos tejidos FFPE de cáncer colorrectal con antigüedad de 11 años. Se empleó un protocolo de conversión con bisulfito alternativo al usual; se usó una DNA polimerasa modificada y una PCR anidada y se optimizó la secuenciación directa del DNA convertido con bisulfito. Este protocolo podría ser aplicado para determinar estados de metilación en otros genes y tipos de cáncer en tejidos FFPE.

Abstract Alterations in the methylation of CpG dinucleotides in promoter regions is one of the epigenetic mechanisms involved in cancer that has potential use as a biomarker. Its evaluation from formalin-fixed and paraffin-embedded (FFPE) tissues represents a great challenge given the partial degradation, crosslinking, and low amounts of the obtained DNA. In this technical note we describe a protocol for the study of the methylation status of the distal promoter of the K-RAS proto-oncogene from several samples obtained from two 11-years old FFPE tissues of colorectal cancer. An alternative bisulfite conversion protocol to the usual one was used; a modified DNA polymerase and a nested PCR were used and the direct sequencing of the converted DNA with bisulfite was optimized. This protocol could be applied to determine methylation states in other genes and types of cancer.

Comparative study of rapid paraffin section versus frozen section of histopathological evaluations of donor organ / 中华器官移植杂志

Objective:To explore the advantages and disadvantages of frozen section versus rapid paraffin section in the evaluations of donor organ.Methods:Five cases of donor liver and 8 cases of discarded donor kidney were collected from 2017 to 2021.Tissues were harvested and prepared by frozen section, rapid paraffin section and normal paraffin section.After hematoxylin-eosin (H&E) staining, the specimens of donor kidney/liver were evaluated by differential histopathological structures and donor quality scoring system.Results:Rapid paraffin section was similar to normal paraffin section in reflecting the proportion of glomerulosclerosis (18.6%±22.3%), arteriolar hyaline degeneration (43.7%±23.8%) and arteriolar stenosis (47.9%±29%). The proportion of glomerulosclerosis (0.8%±2.2%), arteriolar hyaline degeneration (4.9%±7.4%) and arteriolar stenosis (5.3%±7.5%) were lower in frozen sections than those in rapid paraffin sections.The diagnoses of hydropic degeneration and necrosis in donor liver were more accurate in rapid paraffin section.Conclusions:Rapid paraffin section is superior to frozen section in observing histopathological changes under microscope.Scoring of donor organ is more precise according to rapid paraffin section.

A Comparative Study On The Effect Of Paraffin Wax Therapy With Manual Mobilization Versus Ultrasound Therapy With Manual Mobilization On Pain, Range Of Motion And Functional Status Of Shoulder In Diabetic Subjects With Adhesive Capsulitis

Background: The shoulder is a complex joint that plays a crucial role in many activities of daily living. Adhesive capsulitis is defined as a painful and disabling condition in which the capsule and the connective tissue surrounding the glenohumeral (GH) joint becomes inflamed, leading to restriction of range of motion in the joint. A wide range of physical therapy interventions are used to improve the joint range of motion in subjects with adhesive capsulitis. Earlier, isolated studies were done on the effectiveness of paraffin wax therapy and ultrasound therapy in subjects with adhesive capsulitis. The need of the present study was to compare the effect of paraffin wax therapy with manual mobilization and ultrasound therapy with manual mobilization on pain, shoulder ROM and functional status in diabetic subjects with adhesive capsulitis. Methods: Subjects for the study were recruited from the outpatient department of endocrinology and physiotherapy. The size of the sample is 80 (eighty subjects) 40 in each group, and study design is an experimental study design (Non-randomized open-label study). The pretreatment values of pain and functional status of the shoulder were assessed using Shoulder Pain and Disability Index (SPADI) scale and ROM was assessed using a universal goniometer. The subjects in the paraffin wax therapy group received wax therapy for 10 minutes and undergone manual mobilization. The subjects in the ultrasound therapy group received ultrasound therapy (3MHz) for 10 minutes and undergone manual mobilization. The treatment was given for 3 sessions per week for 6 weeks and post-treatment values were noted for the subjects of both the groups. Results: There was an improvement of ROM of external rotation, pain & functional status of the shoulder in subjects who underwent paraffin wax therapy with mobilization and ultrasound therapy with mobilization. When comparing both the groups, the subjects who underwent paraffin wax therapy with mobilization have better outcomes than ultrasound therapy with the mobilization group (p< 0.05). Conclusion: There was an improvement of ROM of external rotation, pain & functional status of the shoulder in subjects who underwent paraffin wax therapy with mobilization and ultrasound therapy with mobilization. When comparing both the groups, the subjects who underwent paraffin wax therapy with mobilization have better outcomes than ultrasound therapy with the mobilization group.

Xylene-free staining: Is it possible and practical?

Good paraffin sections are key to correct histopathological diagnosis. Xylene is hazardous to health, expensive, and difficult to dispose. Various substitutes have been tried without success. We aimed to examine if 1.7% dishwasher soap (DWS) aqueous solution and refined mineral oil (RMO) for deparaffinization can replace xylene. Fifty tissue blocks consisting of benign and malignant lesions were processed using xylene (A), 1.7% DWS (B), and RMO (C). Each section was evaluated, scored as 0 (inadequate) and 1 (adequate) by two independent pathologists who were blinded to agent used. Following criteria were considered: nuclear staining, cytoplasmic staining, clarity, crispness, and uniformity. Total score of <2 was graded as inadequate for diagnosis and 3–5 as adequate. Statistical analysis was done using the SPSS software by applying chi-square test. Among three methods, B had the best scores in adequacy for cytoplasmic staining (P = 0.001), clarity (P = 0.004), and crispness (P = 0.003). About 1.7% DWS and RMO were found to be effective methods for deparaffinization and can replace xylene.

Effect of Liquid Paraffin Gauze Dressing In Burn Wounds: A Prospective Study

Introduction: Burn injury causes a considerable amount of disability, prolonged hospital stay, and burden on the public healthsector. Main requirement in burn wound management is an economical, easy to apply, readily available dressing, or method ofcoverage that will provide good pain relief, protect the wound from infection, promote healing, prevent heat and fluid loss, beelastic, non-antigenic, and adhere well to the wound while waiting for spontaneous epithelialization of superficial partial thicknessburns. The sterilized paraffin gauze dressing is non-adherent and non-allergenic and helps in speedy recovery of burn wounds.Materials and Methods: A prospective study of 90 patients with partial thickness burns who were salvageable (≤40% bodysurface area), admitted to Burn unit of Shyam Shah Medical College and associated Sanjay Gandhi Memorial Hospital,Rewa from June 1, 2017, to May 31, 2018. The autoclaved liquid paraffin gauze was applied over burn wound. Patients wereassessed on the basis of subsidence of pain, time of epithelialization if occurred after liquid paraffin gauze dressing. Patients’blood investigations were noted and the assessment of the effect of hemoglobin (anemia) and platelet counts in burn woundhealing in terms of mean epithelialization time were done.Results: Mean epithelialization time was 16 days. In 25% of cases epithelialization developed in 10–12 days. Post-burn painsubsided in 4–6 days in maximum in 54.44% cases. Mild and moderate anemia had no significant effect on wound healingtime (mean epithelialization time). Patient with less than normal platelet counts (<1.5 lakh/cumm) had more epithelializationtime and with normal platelet count had less epithelialization time. 15 patients developed complications and most commoncomplication was hyper granulation (11.11%).Conclusion: Burn wounds pose a great burden on health-care infrastructure and burn units. We can conclude that liquid paraffingauze dressing has good patient acceptability and less painful, it is easily available and relatively less expensive. In developingand resource-poor countries, most of the patients are from the rural background so these patients will need a dressing that isrelatively less expensive and easily available such as liquid paraffin gauze dressing.

Surgical Techniques for Correction of Penile Paraffinoma

@#Background: Penile augmentation using injection of a foreign body into penile skin was mainly performed by non-medical personnel. Majority of these patients end up with complication of an abnormal mass formation known as penile paraffinoma. Methods: We described three different surgical techniques for correction of penile paraffinoma based on our single-centre experience. Informed consents were obtained from patients whose photographs were taken during the operation step. Results: In general, three patients had simple excision biopsy with primary suturing, four patients underwent single stage excision of circumferential granuloma with bilateral scrotal skin flap reconstruction and one patient experienced dual stage procedure. Three of them were injected with paraffin, one with silicone and the remaining four were unable to identify the substance used. All patients successfully underwent the surgical procedure and four of them had minor postoperative surgical site infection and wound gapping. Conclusion: All patients recovered well and the mean International Index of Erectile Function (IIEF-5) score obtained was 24.25. In our experience, excision biopsy was adequate for focal mass and reconstructive surgery using bilateral scrotal flap was suitable for circumferential mass.

Multistaining Optimization for Epstein-Barr Virus–Encoded RNA In Situ Hybridization and Immunohistochemistry of Formalin-Fixed Paraffin-Embedded Tissues Using an Automated Immunostainer

BACKGROUND: Single staining is commonly performed for practical pathologic diagnoses. However, this method is limited in its ability to specify cellular morphology and immunophenotype and often requires consumption of limited tissue. This study aimed to describe an optimized protocol for multiple in situ hybridization (ISH) and immunohistochemistry (IHC). METHODS: The quality of multistaining was evaluated by carefully changing each step of ISH and IHC in an angioimmunoblastic T-cell lymphoma (AITL) case on a Ventana BenchMark XT automated immunostainer. The optimized protocols were also performed using another immunostainer and in 15 cases of five Epstein-Barr virus (EBV)–associated malignancies using formalin-fixed paraffin-embedded tissue. RESULTS: The quality of various ISH-IHC staining protocols was semi-quantitatively evaluated. The best EBV-encoded RNA (EBER)-ISH/double IHC staining quality, equivalent to single staining, was obtained using the following considerations: initial EBER-ISH application, use of protease and antigen retrieval reagent (cell conditioning 1 [CC1] treatment time was minimized due to impact on tissue quality), additional baking/deparaffinization not needed, and reduced dilution ratio and increased reaction time for primary antibody compared with single immunostaining. Furthermore, shorter second CC1 treatment time yielded better results. Multiple staining was the best quality in another immunostainer and for different types of EBV-associated malignancies when it was performed in the same manner as for the Ventana BenchMark XT as determined for AITL. CONCLUSIONS: EBER-ISH and double IHC could be easily used in clinical practice with currently available automated immunostainers and adjustment of reagent treatment time, dilution ratio, and antibody reaction time.

DNA re-use after immunohistochemical staining of FFPE sections of non-small cell lung cancer / 医学研究生学报

Objective　The purpose of this study was to evaluate the quality of DNA from the formalin-fixed paraffin-embedded (FFPE) specimens of non-small cell lung cancer (NSCLC) after immunohistochemical staining and investigate DNA extraction by immunohistochemical staining of the specimens in small in number or difficult to obtain and the feasibility of related molecular tests.　Methods　We randomly collected 50 FFPE biopsy specimens of NSCLC in our Department of Pathology from June 2017 to December 2017 and sliced each into 12 sections, of which, 6 were directly subjected to DNA extraction (the control group) and the other 6 to immunohistochemical Envision two-step staining for DNA extraction (the experimental group). Then, we detected the mutations of the epidermal growth factor receptor (EGFR), kirsten rat sarcoma viral oncogene (KRAS) and V-raf murine sarcoma viral oncogene homolog B (BRAF) in all the DNAs extracted.　Results　No statistically significant differences were observed between the experimental and control groups in the DNA concentration and purity in the 50 FFPE biopsy specimens of NSCLC (P>0.05). Of the 50 NSCLC FFPE specimens of the experimental group, 20 (40%) showed the mutation of EGFR, 8 (16%) exhibited that of KRAS, and 5 (10%) manifested that of BRAF. In the other 50 specimens of the control group, 33 showed the mutations of EGFR, KRAS and BRAF. A 100% consistency was found in the results of detection between the experimental and control groups (P=0.000, Kappa=1.000).　Conclusion　 High-quality DNA can be extracted after immunohistochemical staining from NSCLC FFPE specimens, especially those small in number or difficult to obtain, and can be used for downstream molecular analysis of target genes, which is a good method for specimen recycling and provides a solution for subsequent molecular test of scarce or difficult-to-obtain clinical samples.

Fixatives Increase the Efficacy of Gel Formulations containing Piper aduncum Linnaeus (Piperales: Piperaceae) Essential Oil as Repellent / Jurnal Sains Kesihatan Malaysia

@#Piper aduncum essential oil exhibit repellency activity and has a potential to be use as an alternative for synthetic repellent such as N,N-diethyl-3- methylbenzamide, (DEET). However, the volatility properties of the essential oil decrease their persistence as a topical repellent. Study has shown that formulation of the essential oil with some fixatives may increase their effectiveness. Therefore, this study was conduct to evaluate the effectiveness of gel formulation containing P. aduncum essential oil with two fixative; vanillin and paraffin oil. Gel formulations containing P. aduncum essential oil with 5% and 10% vanillin and 6% paraffin oil was prepared and tested against Aedes aegypti in laboratory using Standards and Industrial Research Institute of Malaysia (SIRIM) bioassay method. After 240 minute post-application, formulation containing 5% and 10% vanillin was able to provide >70% repellency percentage against mosquito while formulation containing 6% paraffin oil gives <30% repellency percentage. As conclusion, P. aduncum based repellent gel containing 5% and 10% vanillin was able to prolong the effect of P. aduncum essential oil as repellent against dengue vector in laboratory.

Quantitative Expression of RNA from Frozen Organs and Formaldehyde-fixed and Paraffin-embedded Tissues / 法医学杂志

Objective Quantitative analysis and comparison of the expression of ribonucleic acid （RNA） from frozen organs and formaldehyde-fixed and paraffin-embedded （FFPE） tissues. Methods Frozen specimens of human brain, myocardium and liver tissues as well as FFPE samples at different postmortem intervals were collected and mass concentration of RNA was extracted and detected. Reverse transcription-quantitative polymerase chain reaction （RT-qPCR） technology was used to analyze the amplification efficiency and relative expression of each RNA marker. Results The mass concentration and integrity of RNA extracted from FFPE samples were relatively low compared with frozen specimens. The amplification efficiency of RNA markers was related with RNA species and the length of amplification products. Among them, glyceraldehyde-3-phosphate dehydrogenase （GAPDH） and β-actin （ACTB） with relatively long amplification products failed to achieve optimal amplification efficiency, whereas 5S ribosomal RNA （5S rRNA） achieved ideal amplification efficiency and showed quite stable expression across various tissues, therefore it was chosen as internal reference marker. The expression quantity of GAPDH and ACTB in frozen specimens with longer postmortem intervals and in FFPE samples with relatively long amplification products was decreased. The expressions of tissue-specific microRNAs （miRNAs）, GAPDH and ACTB with relatively short amplification products had consistency in the same tissues and FFPE samples. Conclusion Through standardizing the RT-qPCR experiment, selecting the appropriate RNA marker and designing primers of appropriate product length, RNA expression levels of FFPE samples can be accurately quantified.

Comparative Study Of Effectiveness Of Cyanoacrylate Versus Paraffin Gauze Dressing On Split Thickness Skin Graft Donor Site

Object: - The aim of this study was comparison of the results of use of cyanoacrylate versus paraffin gauze dressing on split thickness skin graft donor site. Materials & Methods - This prospective study was conducted on 50 patients who were admitted in surgical wards in tertiary care hospital from December 2016 to July 2018 and underwent split skin grafting. Results- In our study we observed that in cyanoarylate group patients, homeostasis and healing were faster and itching and pain as well as hospital stay was less as compared to paraffin gauze group patients. Conclusion- Use of cyanoacrylate over donor site is very good in comparison with paraffin gauze dressing of donor site in respect to healing, homeostasis and time of hospital stay

Contralateral Internal Mammary Lymphadenopathy Mimicking Metastasis in a Patient with a History of Breast Cancer and Prior Interstitial Mammoplasty by Paraffin Injection: MRI, PET-CT, and Pathological Findings

Foreign body injections into breasts may produce foreign body reactions, fibrosis, and local swelling of involved lymph nodes, which can be misdiagnosed as metastasis or malignancy. Here, the authors report MR imaging, PET-CT imaging, and pathologic findings of contralateral internal mammary lymphadenopathy suspicious of breast cancer metastasis in a 58-year-old woman with history of left breast cancer, and previous interstitial mammoplasty by paraffin injection in both breasts.