Effect of Eosin Solution Exposed to Sunlight against Anopheles Mosquito Larvae

Mosquitoes are a menace for millions of people around the world, they are vectors for destructive microorganisms which cause diseases like Malaria, Dengue fever and Lymphatic filariasis, affecting people in developing countries and areas with tropical climates. Anopheles, a predominant genus transmits malaria, and the World Health Organization have shown that 350,000 lives especially children and pregnant women are lost annually by it. The aim of this study is to determine the effect of eosin solution exposed to sunlight on larvae of Anopheles mosquito. The use of control method was aid at eliminating the larva stages of the mosquito life cycle. A total of one hundred and twenty (120) Anopheles mosquito larvae were harvested using dipper with handle and net from drainages at Eagle Island and Rivers State University both in Port Harcourt. Five different concentrations of Eosin solutions were prepared in volumes of 1000 microlitre (µl), 800µl, 600µl, 400µl, 200µl after a stock solution of 1gram(g) in 100ml and a control, the physicochemical parameters of the solutions were determined using Extech model DO700 measuring instrument. Twenty (20) mosquito larvae, were carefully introduced into each of the concentrations, exposed to sunlight and observed for 24 hours (hrs) for a period of six (6) days for susceptibility. A hundred percent (100%) mortality was recorded in eosin volume of 1000µl and 800µl. The separate solutions of eosin showed significant effects of their concentrations on the Anopheles mosquito larvae of P-value 0.017 at P<0.05. The result obtained for the physicochemical parameters were; pH 5.24, temperature 30.4oC, conductivity 168µS/cm, salinity 0.08%, total dissolved solids 118 milligram per litre(mg/L) and dissolved oxygen was 6.5mg/L for the control. Changes occurred in the values of the dissolved oxygen before and after exposure to sunlight in all the dilutions. The results obtained showed that after 24 hrs, the mortality rate of the larvae increased, indicating that Anopheles mosquito larvae expose to concentrations of eosin solutions results in their mortality within 48 hrs. It may be concluded that this study has provided some evidence of larvicidal effect of eosin solution exposed to sunlight on larvae of Anopheles mosquito.

Histopathological alterations in gills, liver, kidney and muscles of Ictalurus punctatus collected from pollutes areas of River / Alterações histopatológicas em brânquias, fígado, rim e músculos de Ictalurus punctatus coletados em áreas poluídas do rio

Abstract Untreated sewage and industrial wastes from Faisalabad city are disposed to River Chenab through Chakbandi Main Drain (CMD). The present project is planned to investigate the effects of this freshwater pollution on the body of fish Ictalurus punctatus. The specimens of this fish species were collected upstream and downstream of the entrance of CMD into River Chenab. Fish gills, liver, kidney and muscles from dorsolateral regions of fish were subjected to histopathology. Farmed fish and fish from upstream areas were used as control. Fish collected from polluted experimental sites showed significant damage in selected organs. Gill tissues showed an abnormality in the form of an uplifting of the primary epithelium, fusion, vacuolation, hypertrophy, and necrosis. While liver tissues subjected to hepatocytes degeneration, necrosis, mitochondrial granular hepatocyte, and sinusoids dilation. Kidney tissues indicated increased bowmen space and constricted glomerulus and degenerated nephrons. Edema, necrosis, and atrophy were observed in muscle tissues of fish from polluted areas. Fish from the upstream area showed fused gill lamellae, inflammatory cell infiltration, hypertrophy and vacuolation in hepatocytes. Kidney tissues indicated the presence of nuclear tubular cells, destructive renal tubules, hemorrhage, and necrosis at tubular epithelium. Intra myofibril spaces were also observed in muscles. Specimens of control fish indicated no variation in gills, liver, kidney, and muscles. The present study revealed a strong correlation between the degree of tissue damage and environmental contamination. Present findings also compel global warnings to protect our water bodies and fish to rescue the human population.

Resumo O esgoto não tratado e os resíduos industriais da cidade de Faisalabad, no Paquistão, são descartados no Rio Chenab através do dreno principal de Chakbandi (CMD). O presente projeto busca investigar os efeitos dessa poluição de água doce no corpo de peixes Ictalurus punctatus. Os espécimes deste peixe foram coletados a montante e a jusante da entrada do CMD no Rio Chenab. Brânquias, fígado, rim e músculos das regiões dorsolaterais dos peixes foram submetidos à histopatologia. Peixes de criação e peixes de áreas a montante foram utilizados como controle. Peixes coletados em locais experimentais poluídos mostraram danos significativos em órgãos selecionados. Os tecidos branquiais mostraram uma anormalidade na forma de elevação do epitélio primário, fusão, vacuolação, hipertrofia e necrose. Observou-se que os tecidos hepáticos estão sujeitos a degeneração de hepatócitos, necrose, hepatócitos mitocondriais granulares e dilatação de sinusoides. Os tecidos renais indicaram aumento do espaço dos arqueiros, glomérulos contraídos e néfrons degenerados. Edema, necrose e atrofia foram observados nos tecidos musculares de peixes de áreas poluídas. Peixes da área a montante apresentaram lamelas branquiais fundidas, infiltração de células inflamatórias, hipertrofia e vacuolização em hepatócitos. Os tecidos renais indicaram a presença de células tubulares nucleares, túbulos renais destrutivos, hemorragia e necrose no epitélio tubular. Os espaços intramiofibrilas também foram observados nos músculos. Amostras de peixes controle não indicaram variação em brânquias, fígado, rim e músculos. O presente estudo revelou uma forte correlação entre o grau de dano tecidual e a contaminação ambiental. As descobertas atuais também constituem avisos globais para proteger nossos corpos d'água e peixes para resguardar a população humana.

Value of fluorescence staining in histopathological diagnosis of Acanthamoeba keratitis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the application value of fluorescent staining technique in the detection of amoebic pathogens in corneal tissue biopsy, and to apply fluorescent staining technique in the histopathological diagnosis of Acanthamoeba keratitis(AK), comparing the results with those of hemotoxyiln-eosin staining(HE staining)and periodic acid-schiff staining(PAS staining), and analyzing the sensitivity and specificity of these three staining methods.<p>METHODS:Specimens of infected corneal tissue were collected from 74 cases(75 eyes), and then they were divided into an AK group and a non-Acanthamoeba keratitis(NAK)group based on the results of corneal scraping, culture and histopathological diagnosis. The tissues of consecutive sections were stained with HE staining, PAS staining and fluorescence respectively, and the sensitivity and specificity of the three staining methods for the diagnosis of AK were analyzed. Area under the curve(AUC)was calculated using the receiver operating characteristic(ROC)curve. Further analysis was performed to count the number of Acanthamoeba pathogens found by the three staining methods under the same magnification field of view at the same site, and to clarify the diagnostic value of fluorescent staining technique for AK.<p>RESULTS: The sensitivity of HE staining was 69%(27/39)with a specificity of 92%; the sensitivity of PAS staining was 62%(24/39)with a specificity of 97%, and the sensitivity of fluorescent staining was 95%(37/39)with a specificity of 97%. There were differences in the sensitivity of the three staining methods for the diagnosis of AK(χ2=19.857, <i>P</i><0.001), and pairwise comparison revealed that the differences between HE staining and fluorescent staining, PAS staining and fluorescent staining for the diagnosis of AK were statistically significant(<i>P</i>=0.003,<0.001), while the difference in sensitivity between HE staining and PAS staining for the diagnosis of AK was not statistically significant(<i>P</i>=0.978). The maximum AUC was 0.960 for fluorescence staining, followed by 0.804 for HE staining and 0.794 for PAS staining, respectively. The median number of amoeba cysts detected by HE staining, PAS staining and fluorescent staining at the same site under the same magnification field of view was 4(0, 11), 2(0, 9)and 12(3, 33), respectively(χ2=56.561, <i>P</i><0.001). Pairwise comparison revealed that the differences in the number of amoeba cysts found by HE staining and fluorescence staining, PAS staining and fluorescence staining were statistically significant(<i>P</i><0.001), while the difference in the number of amoeba cysts found by HE staining and PAS staining was not statistically significant(<i>P</i>=0.210). Fluorescently stained histopathological sections make it easier to identify amoebic pathogens.<p>CONCLUSION:Fluorescent staining technique is more sensitive to histopathological diagnosis of AK than HE staining and PAS staining, which can significantly improve the positive rate of detection of amoebic pathogens.

MicroRNA-141-3p expression in serum of patients with cerebral hemorrhage and its mechanism / 解剖学报

Objective To analyze the expression level of microRNA-141-3p (miR-141-3) in patients with intracerebral hemorrhage (ICH), and explore the effect and mechanism of miR-141-3p on cerebral hemorrhage injury in rats. Methods Forty patients with ICH and 40 healthy controls in total were enrolled in this study. The expression of miR- 141-3p in peripheral blood serum was determined by the Real-time PCR method. The target relationship between miR-141- 3p and NOD-like receptor 3 (NLRP3) 3′ UTR was confirmed by dual luciferase reporter assay. miR-141-3p agonist and agonist NC were injected into rats via the lateral ventricle, respectively. On day 7 after treatment, the neurological function score was evaluated, and then all rats were killed to obtain brain tissue. Brain water content was examined by the dried and wet mass. HE staining was conducted to observe the pathological changes of cerebral tissue. The mRNA expressions of NLRP3 and miR-141-3p were detected by Real-time PCR. The protein expression of interleukin (IL)-lβ, IL-6 and IL-18 were detected by Western blotting analysis. Results The expression of miR-141-3p in serum of ICH patients was significantly down-regulated compared to healthy controls and negatively correlated with the severity of edema around the hematoma [(0.068±0.038) vs (0.520±0.028), t = 15.93, P<0.001; r =-0.8948, -0.9434 to-0.8087, P<0.001 ]. The result of luciferase reporter assay showed that miR-141-3p was related to the regulation of NLRP3 gene expression. The relative expression levels of miR-141-3p in agonist group were significantly higher than those in the agonist NC group (P< 0.001), while the expression levels of NLRP3, IL-lβ, IL-6 and IL-18 were significantly lower than those in the agonist NC group (P< 0.001). Compared with the agonist NC group, the cerebral water content reduced significantly (P< 0.001), and the neurological function score was significantly improved on the day 7 after treatment in agonist group (P< 0.001). The result of HE staining showed that injection of miR-141-3p in ICH rats could reduced the severity of edema around the hematoma. Conclusion MiR-141-3p alleviates ICH-induced inflammatory injury in rat possibly by modulating miR-141-3p.

Efficacy of Eosin Dye Removal by Peanut Shell Agrowaste Adsorbent

Abstract Peanut shell (PS) which is an excessive waste-product from agricultural processes, it can be recycled to a natural adsorbent for example it uses as removal dyes. Synthetic dye effluent without improperly discharged from industries to the river cause wastewater and damage to living organisms, especially, anionic dyes are difficult removed by conventional treatments such as biological, chemical, oxidation, and physical-filtration. However, an adsorption treatment is widely used for decolorization of dyes and give the best results for removal of various types of dissolved coloring materials. This research was used Eosin Y (EO) for the anionic model of dyeing wastewater and used PS for agrowaste adsorbent. The purpose of this study was investigated the efficiency adsorption of EO removal by PS. This efficiency adsorption was measured by different PS dosages, contact times, adsorbate concentration and equilibrium data. The results can be concluded that the PS had the efficiency adsorption of EO removal due to the equilibrium adsorption capacity (qe) and the highest dose of PS were balanced to adsorption of dye. The highest EO removal percentage was found in 87.7%, the qe was 0.351 mg g-1 and can adsorb from 10 mg L-1 to 1.23 mg L-1 in 25 g L-1 of PS dose at 30 minutes. In addition, the PS structure was found in multi-layer and many porous which is suitable for adsorbent. The morphological examination of PS was shown before and after adsorption that not changed. Therefore, PS might be an alternative choice for removal dye, and be used for the recycle adsorbent agrowaste as a commercial product for adding their values.

Comparison of HE and Giemsa staining in diagnosis of bacterial and allergic conjunctivitis in children / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To explore the diagnostic effect of hematoxylin-eosin staining(HE)and Giemsa staining in the diagnosis of bacterial and allergic conjunctivitis in children. <p>METHODS:Totally 422 children with conjunctivitis diagnosed by conjunctivitis from the ophthalmology department of our hospital during 2016-10/2019-10 as the research objects. HE and Giemsa staining methods were used to stain the conjunctival scratches, and the staining results were used to diagnose bacterial/allergic conjunctivitis. Observe the positive detection rate of the two staining results for bacterial/allergic conjunctivitis and the staining situation. <p>RESULTS: The positive rate(33.0%)and coincidence rate(63.6%)of HE staining for the diagnosis of bacterial conjunctivitis were significantly lower than Giemsa staining(90.7% and 88.8%, <i>P</i><0.001), while the positive rate of allergic conjunctivitis was not significantly different 90.8% <i>vs </i>87.2%, <i>P</i>>0.05).<p>CONCLUSION: The Giemsa staining method can accurately diagnose bacterial conjunctivitis in children and the method is simple. Both HE and Giemsa staining methods have good diagnostic effects on allergic conjunctivitis, which can provide a basis for improving the clinical diagnosis efficiency and early treatment options.

Effect of miR-155 on acute lung injury in burned rats: Changes of nuclear factor-kappa B pathway / 中国组织工程研究

BACKGROUND: Currently, studies have focused on the role and mechanism of nuclear factor-kappa B pathway in the pathological process of acute lung injury in burned rats, such as the targeting inhibition of kB kinase by miR-155, which further weakens the activity of nuclear factor-KB and plays a role in acute lung injury in burned rats. However, there are still some pathological mechanisms to be studied and confirmed. OBJECTIVE: To investigate the effect of miR-155 on acute lung injury in burned rats through nuclear factor-KB pathway. METHODS: The rat models of acute lung injury were established by warm water bath simulating bum injury. The burned rats were divided into acute lung injury, miR-155-mimics and miR-155-inhibitor groups. After fluid resuscitation, the rats in the miR-155-mimics and miR-155-inhibitor groups were injected into the tail vein of 5 |_iL of miR-155-mimics and miR-155-inhibitions, respectively. The expression levels of tumor necrosis factor-a and interleukin-1 p in bronchoalveolar lavage fluid were detected by ELISA. The lung morphology in the three groups was observed by hematoxylin-eosin staining. The protein expression levels of nuclear factor-KB and cyclooxygenase 2 were evaluated by western blot assay. The nuclear factor-KB protein in lung tissues was detected by immunohistochemistry. RESULTS AND CONCLUSION: (1) The results of hematoxylin-eosin staining showed that the severity of lung injury in the miR-155-inhibitor group, acute lung injury group and the miR-155-mimics group was increased gradually (P < 0.05). (2) ELISA results showed that compared with the acute lung injury group, the expression levels of tumor necrosis factor-a and interleukin-1 p were increased in the miR-155-mimics group (P < 0.05), and decreased in the miR-155-inhibitor group (P < 0.05). (3) Western blot assay results showed that compared with the acute lung injury group, the expression levels of nuclear factor-KB and cyclooxygenase 2 proteins were increased in the miR-155-mimics group (P < 0.05), and decreased in the miR-155-inhibitor group (P < 0.05). (4) Immunohistochemical results showed that the expression level of nuclear factor-KB was increased in the miR-155-inhibitor group, which was dark brown. The expression of nuclear factor-KB in cytoplasm and nucleus of neutrophils, mononuclear macrophages, alveolar epithelial cells was the most obvious. (5) These results indicate that in lung tissue cells, decreased miR-155 can down-regulate nuclear factor-KB activity, which reduces the inflammatory response of the lung between the damaged tissue. The study was approved by the Laboratory Animal Ethics Committee of the First People’s Hospital of Neijiang, approval No. 1801270.

Lycium barbarum polysaccharide intervenes with CD151 and matrix metalloproteinase 3 in the articular cartilage in a rabbit model of knee osteoarthritis / 中国组织工程研究

BACKGROUND: Studies have shown that Lycium barbarum polysaccharide (LBP) has the functions of anti-aging, nerve protection, anti-fatigue, blood sugar control, anti-oxidation, and anti-tumor. It may have some protective effects against osteoarthritis of the knee, but have been rarely reported. CD151 and matrix metalloproteinase 3 (MMP-3) are two common cytokines for assessing knee osteoarthritis. OBJECTIVE: To observe the effect of LBP on the expression of CD151 and MMP-3 in rabbit osteoarthritis. METHODS: Sixty-four healthy 6-month-old white rabbits were randomly divided into four groups: blank group, model group, LBP group and normal saline group. Animal models of knee osteoarthritis were made using Hulth method in the rabbits except those in the blank group. The rats in the LBP and normal saline groups were fed with normal dose of LBP and normal saline for 4 weeks, and then the articular cartilage tissues were taken from the affected side at 12 weeks after modeling. The morphological changes of the articular cartilage were observed by hematoxylin-eosin staining. The expression levels and spatial distribution of CD151 and MMP-3 in articular cartilage was observed by immunohistochemical staining and western blot. Ethic approval was given by the People’s Hospital of Ningxia Hui Autonomous Region (approval No. 2014-30817). RESULTS AND CONCLUSION: immunohistochemistry staining and western blot results showed that the absorbance values and protein expression of MMP-3 and CD-151 were significantly lower in the LBP group than the normal saline and model groups (P < 0.05). Therefore, the expression of CD151 and MMP-3 in the articular cartilage of osteoarthritis was increased, and LBP could inhibit the expression of CD151 and MMP-3 in osteoarthritis, so as to slow down the occurrence of osteoarthritis.

Effect of spinal cord decellularized scaffold on the repair of spinal cord defects in rats / 解剖学报

Objective Exploring the effect of spinal cord decellularized scaffold on spinal cord defects and observing the behavior and regeneration of rats after operation. Methods The spinal cords of 30 SD rats were treated with 3% Triton X-100 and 2% sodium deoxycholate on oscillator. The cell residue and the spatial structure of the tissue were compared before and after treatment, in order to understand the tissue structure of the stent itself. 90 SD rats were randomly divided into control group, simple injury group and stent transplantation group. Excision of the spinal cord 9-10 segments in the simple injury group and the stent graft group the acellular scaffold was transplanted to the stent graft group. Behavioral scores were observed postoperatively. At 4, 8, and 12 weeks, the spinal cords of the injured part of the rats were taken for HE staining and immunofluorescence detection of nerve regeneration-related proteins. Results After decellularization of the spinal cord, the nerve cells and axons were completely removed, and the extracellular matrix of the spinal cord was preserved. Scanning electron microscopy revealed that the scaffold retained a certain porous network scaffold structure. In the experiment of decellularized scaffold in vivo, the Basso-Beattie-Bresnahan(BBB) score showed that the recovery of hindlimb motor function in rats with decellularized scaffolds was better than that in rats with simple injury. HE staining showed that the decellularized scaffold could fill the defect of the spinal cord segment and accelerate the repair process of the injured spinal cord. Immunofluorescence showed that there was a certain axonal regeneration in the injured part of the stent transplantation group. Conclusion The spinal cord decellularized scaffold retains the extracellular matrix and has a certain spatial structure, which can accelerate the process of spinal cord defect repair to a certain extent, and has a certain promoting effect on nerve regeneration.

Improvement program on pretreatment of acid decalcified tissue in hematoxylin-eosin staining / 华西口腔医学杂志

OBJECTIVE@#To explore the treatment conditions of acid decalcified specimens and improve the poor quality of sections and unclear structure of hematoxylin-eosin (HE) staining caused by the change in pH in tooth and hard tissue after acid decalcification.@*METHODS@#A total of 20 cases of oral pathological specimens that contain hard tissues were decalcified and treated with routine treatment, concentrated ammonia water immersion treatment, and saturated lithium carbonate solution immersion treatment. The quality and HE staining effects of hard tissue sections treated with different methods were compared.@*RESULTS@#Compared with routine treatment, lithium carbonate saturated solution treatment showed complete sections. Hematoxylin is strongly stained, the nucleus is clear, and the cytoplasm is bright.@*CONCLUSIONS@#Soaking acid decalcified specimens in lithium carbonate saturated solution before embedding in dehydration can neutralize the acidic environment of the tissue. The quality of sections and HE staining effect are improved and are suitable for the pretreatment of acid decalcified tissue samples of oral pathology.

Identificación de Helicobacter pylori por medio de la coloración especial de Warthin-Starry en biopsias de pacientes con gastritis crónica folicular, previamente negativas en la tinción de hematoxilina-eosina / Warthin-Starry stain identification of Helicobacter pylori in biopsies of patients who previously tested negative in hematoxylin-eosin staining for follicular gastritis

Resumen Las técnicas empleadas para la detección del Helicobacter pylori (H. pylori) son no invasivas e invasivas. En estas últimas, la presencia del H. pylori se determina a partir de la tinción de hematoxilina-eosina (HE), prueba rutinaria, mientras que en pocas ocasiones se aplica la tinción de Warthin-Starry (WS) como coloración especial. Objetivo: identificar la presencia de H. pylori por medio de la coloración especial de la WS en biopsias de pacientes con gastritis crónica folicular, previamente negativas en la tinción HE. Materiales y métodos: se desarrolló un estudio de tipo descriptivo transversal, en un período de 12 meses. Se tomaron los bloques de parafina de las muestras de la mucosa gástrica de pacientes con diagnóstico de gastritis crónica e hiperplasia folicular. Además, se extrajo un corte histológico del mismo bloque, al cual se le aplicó HE y se determinó la presencia o ausencia de H. pylori. Así, de estar ausente, se tomó del mismo bloque un corte adicional y se aplicó WS. Esto se evaluó con el fin de identificar la existencia o no del bacilo. Resultados: se recolectaron 314 muestras; 209 fueron negativas y 105 fueron positivas para HE. El 45 % (94) de estas muestras fueron positivas respecto a la presencia del bacilo, al aplicar la segunda coloración, y el 55 % (115) de las muestras persistieron negativas. Conclusión: el hallazgo de H. pylori es significativamente alto al aplicar la coloración de WS a muestras cuyo estudio histológico evidenció la ausencia del bacilo en biopsias de la mucosa gástrica, especialmente en muestras con escasa cantidad de bacterias.

Abstract Non-invasive and invasive techniques can be used for detection of Helicobacter pylori. An invasive technique identifies the bacteria through routine hematoxylin-eosin staining. Warthin-Starry stain is rarely used. Objective: Our objective was to identify H. pylori by Warthin-Starry staining of patient's biopsies with chronic follicular gastritis who had previously tested negative in hematoxylin-eosin staining. Materials and methods: This is a descriptive, cross-sectional descriptive study that was carried out over a period of 12 months. The study examined paraffin blocks of samples taken from the gastric mucosa of patients diagnosed with chronic gastritis and follicular hyperplasia. A histological section was extracted from a block and tested with hematoxylin-eosin staining for the presence or absence of H. pylori. If absent, an additional cut was taken from the same block and Warthin-Starry staining was used to retest for the presence of the bacteria. Results: Of the 314 samples collected, 209 tested negative, and 105 tested positive for H. pylori when hematoxylin-eosin staining was used. Of the 209 negative samples, 45% (94) tested positive when Warthin Starry stain was used, and 55% (115) still tested negative. Conclusion: Findings of H. pylori are significantly higher when Warthin Starry stain was used to test samples whose previous histological study had evidenced an absence of the bacillus, especially in samples with a small amount of bacteria.

A Prospective Study of Histomorphological Spectrum of Biopsy Confirmed Skin Adenexal Tumors in a Tertiary Care Centre at Bikaner Region

Introduction: The adenexa are part of skin andcomprised of sebaceous glands, sweat glands and hairfollicles. Skin adenexal tumors are rare in occurrenceand precise classification of these neoplasms is difficult.Benign tumors are more common than malignanttumors. Current study aimed to know the histopathologyof skin adenexal neoplasm and to correlate with age,gender, loacation and type of differentiation.Material and methods: A prospective study of 57histopathologically confirmed cases of skin adenexaltumors was carried out in Department of Pathology. Inthis study biopsies were received in 10% formalin andstained by routine haematoxylin and eosin stain. Nonneoplastic conditions were excluded from the study.Results: Out of the 57 cases of skin adenexal tumorsstudied 55 (96.5% ) were benign and 2 (3.5% ) weremalignant with a male: female ratio of 1:1.3. The mostfrequent group of tumors were of eccrine/apocrineorigin 28/57 (49.1%) followed by follicular origin(38.6%) 22/57 and sebaceous origin 7/57 (12.9%).Pilomatrixoma was the most common benign tumorand sebaceous carcinoma was the only malignant tumorencountered in the study. Most common age groupaffected range from 41-60 years and mean age observedwas 45 years. Head and neck (47.5%, 27/57) was themost common site involved in both males and femaleswith a predominance in the facial region.Conclusion: Skin adenexal tumors (SAT) are very rareand the classification of these tumours is complex. Thesetumors are usually missed clinically and histopathologyproves to be the gold standard for diagnosis of theseneoplasms.

Tetrachromic VOF/Masson's trichrome/H and E stains: Unmasking their usability in differential stromal hard tissue staining

Background: Identification and differentiation of stromal hard tissue components is a challenging task. Numerous methods of demonstrating these components have been worked upon in the past. Although some of the methods have been successful, there are many drawbacks of employing them. The need of the hour, therefore is to develop and use a simple, rapid and cost-effective method of identifying stromal hard tissues as they may signify an important change in the diagnosis of the pathology. Our aim is therefore to observe the usability of tetrachromic VOF stain over Hematoxylin and Eosin and Masson's Trichrome in routinely encountered head and neck pathologies. Materials and Method: Routine cases such as Central and peripheral ossifying fibromas, osteomas, giant cell granulomas, osteomyelitis and malignancies like osteosarcomas were retrieved from the department archives and 3 sections from each block were prepared to stain with H and E, Masson's trichrome and modified tetrachromic VOF stains respectively using standard staining protocol. Results: Tetrachromic VOF takes an upper hand in stromal hard tissue differentiation irrespective of the pathology. Conclusion: Modified tetrachromic VOF is simple, cost-effective method and can be employed for diagnosis of cases with hard tissue differentiation within the stroma on routine basis.

Comparison Of Morphological Characteristics Of Pituitary Gland Between The Different Age Groups

Background: The pituitary gland (PTG) size, shape will change according to the age in response to the changes inthe hormonal environment. Hence care should be taken while evaluating the PTG disorders. This present studyconducted to evaluate the morphological changes in PTG with relation to age.Materials and Methods: This study was conducted in the Department of Anatomy was approved by InstitutionalHuman Ethics Committee. A total of 73 PTG specimens were included in this study. They are divided in to sixgroups based on the age. G-I (Foetus), G-II (1-10 Y), G-III (11-30 Y), G-IV (31-50 Y), G-V (51-70 Y) and G-VI(Above 71Y). All the specimens were subjected for H&E stain. The slides were observed for morphological changes. The datawas expressed in MEAN±SD and Statistical Package for Social Sciences (SPSS 16.0) version used for analysis.Results: More number of males was in group-V and females in group-IV. Pars intermedia had maximum thicknessin foetal life. Basophilic zone was not seen in foetal life but it is more prominent in other age groups. Cellularityincreased as age progress. Pars anterior and nervosa showed more vascularity compared to interior. As ageprogress this vascularity is decreased.Conclusion: From the study observations it can be concluded that as age progress there is a significant changesin the PTG morphology. Knowledge about these changes can useful for the diagnosis and treatment of variousdisorders of PTG.

Effect and mechanism of essential oil from Zanthoxylum bungeanum in microemulsion gel preparation on percutaneous delivery of complex components / 中国中药杂志

This study aimed to investigate the transdermal enhancing effect of essential oil from Zanthoxylum bungeanum(Z. bungeanum oil) in microemulsion gel(ZO-ME-gel) on permeation of different components,and reveal the transdermal enhancing mechanism of ZO-ME-gel. A series of components with different log P values were selected as model drugs and encapsulated in ZO-ME-gel to simplify and characterize the complex components of traditional Chinese medicine. The transdermal behavior of the model drugs was further examined using the improved Franz diffusion cell method. Then attenuated total reflection Fourier transform infrared spectroscopy(ATR-FTIR),differential scanning calorimetry(DSC) studies and hematoxylin-eosin(HE) staining were used to investigate the effects of Z. bungeanum oil and ZO-ME-gel on keratin,intercellular lipids and microstructure of the stratum corneum(SC). The results showed that Z. bungeanum oil and ZO-ME-gel had a good transdermal enhancing effect on both hydrophilic and lipophilic drugs,and the best effect was achieved when log P value was-0. 5. The transdermal enhancing mechanism of Z. bungeanum oil and ZO-ME-gel was related to affecting the order of the SC lipids,changing lipid fluidity and protein conformation,and disrupting the integrity of the SC structure. 5% Z. bungeanum oil had greater transdermal enhancing effect and destruction of SC structure than ZO-ME-gel. These results suggested that Z. bungeanum oil loaded in microemulsion gel still had a good transdermal enhancing effect although the effect was not as great as Z. bungeanum oil itself,in addition,ZO-ME-gel was less irritating to the skin and safer to use,which had a guiding role in the development and clinical application of Z. bungeanum oil-containing traditional Chinese medicine topical preparations.

A point of confusion for embryologists in the identification of viable spermatozoa by the eosin-nigrosin test / 대한생식의학회지

A viable spermatozoon is a prerequisite for fertilization in intracytoplasmic sperm injection (ICSI). Thus, it is crucial to select viable but immotile spermatozoa on the day of ICSI. We report conflicting results in the identification of viable but immotile spermatozoa between the eosin-nigrosin staining and the laser test, which resulted in confusion for embryologists during assisted reproductive technology (ART). Three patients’ semen samples that showed no motile spermatozoa are described in this report. To identify viable spermatozoa, we used both the eosin-nigrosin test and the laser test for each sample, and repeated the semen analysis twice in each patient. Viable but immotile spermatozoa selected by the laser test were used for ICSI. Viable spermatozoa were detected by both the eosin-nigrosin and laser tests in two patients (case 1, 95.00% vs. 24.21% and 92.68% vs. 22.22%; case 2, 41.18% vs. 23.48% and 39.81% vs. 22.52%), indicating consistent results between the two methods. In the third patient, the eosin-nigrosin test yielded viability rates of 20.75% and 19.14%, while the result of the laser test was 0%. Thus, testicular aspiration was performed to collect viable sperm from this patient. Normal fertilization was achieved after the injection of viable but immotile spermatozoa selected from these patients by the laser test, resulting in the birth of two healthy babies. Our study documents a case where the eosin-nigrosin test showed a limitation in identifying viable but immotile spermatozoa for ART, while the laser test may overcome this limitation. Larger samples may be required to corroborate the clinical value of the laser test.

A rapid pathological preparation method for composite material observation / 中国修复重建外科杂志

Objective: To explore a simple and rapid pathological slices method to observe the porous structure and the composition distribution of composite materials. Methods: Taking polyurethane/small intestinal submucosa (PU/SIS) composite as an example, PU/SIS was OCT-embedded and sliced into sections by frozen section technology, after which general observation of the section integrity was carried out. After dyed with water-soluble eosin in alcoholic solution, the staining effect and the porous structure of the composite were observed under light field microscope. Sections were sealed with five different sealing methods. Group A: sealing piece using glycerogelatin method; group B: anhydrous alcohol dehydration→transparency using TO transparent reagent→sealing piece using neutral quick drying glue; group C: color separation using deionized water→air-drying→sealing piece using neutral quick drying glue; group D: air-drying→transparency using TO transparent reagent→sealing piece using neutral quick drying glue; group E: air-drying→sealing piece using neutral quick drying glue. Then, the morphology and the components distribution of the composite were observed under light field microscope, and the simple and feasible method was selected as optimum method. Results: From general observation, the frozen section of the PU/SIS composite, which was 6 μm in thickness, was complete and continuous. Although the outline of the material and the porous structure in the sections could be observed clearly under light field microscope, the two components still could not be identified by using eosin staining method. After sealing piece, the material components in groups A, B, and C still could not be identified or be dissolved and deformed; the morphology of the material in groups D and E were preserved and the two components in the composite were clearly visible. Conclusion: The morphology and the components distribution of PU/SIS frozen sections can be characterized after soluble eosin staining and neutral quick drying glue sealing.

Determination of mexiletine hydrochloride in drugs by eosin Y Rayleigh light scattering / 国际药学研究杂志

Objective: To establish a simple,rapid and novel Rayleigh light scattering(RLS)method for rapid determination of mexiletine hydrochloride in drugs. Methods: In the presence of acid Tris-HCl medium and cetylpyri- dinium bromide,eosin Y reacted with mexiletine hydrochloride to form a ternary ion association complex with two charac- teristic scattering peaks by electrostatic attraction. The detection wavelengths were 368 and 586 nm. There was a linear relationship between the mexiletine hydrochloride concentration in a certain range and the Rayleigh light scattering en- hancement intensity(ΔIRLS)of the association complex. Single-wavelength Rayleigh scattering(SWO-RLS)method or dualwavelength Rayleigh light scattering(DWO-RLS)method was used to determine the content of mexiletine hydrochloride, and the mexiletine hydrochloride content was calculated according to the regression equation of standard curve. Results: The linear ranges of mexiletine hydrochloride were 0.005-0.65 mg/L(SWO-RLS method,368 nm),0.004-0.65 mg/L (SWO-RLS method,586 nm)and 0.004-0.65 mg/L(DWO-RLS method,368 nm+586 nm),respectisely. Detection lim- its were 0.0033(SWO-RLS method,368 nm),0.0040(SWO-RLS method,586 nm)and 0.0018 mg/L(DWO-RLS method, 368 nm+586 nm),respectisely. The recovery and relative standard deviation(RSD,n=5)for a SWO-RLS method were 98.6-103% and 1.4-1.8%,respectively(SWO-RLS method,368 nm). Conclusion: The method is simple,rapid, highly sensitive and high selectie.

Comparing The Efficacy Of Dishwash Solution, Diluted Lemon Water, Coconut Oil And Xylene As Deparaffinizing Agents For Hematoxylin And Eosin Staining Procedure

Aim: To compare the efficacy of dish wash solution, diluted lemon water, coconut oil and xylene as deparaffinizingagents for hematoxylin and eosin staining procedure.Objective: The objective is to find eco-friendly deparaffinizing agents like dish wash solution, diluted lemonwater and coconut oil as substitute to xylene and comparing the staining characteristics of each individualdeparaffinizing agent with Xylene.Materials and Methods: The study comprised of paraffin embedded 45 blocks of various tissues. Each block offour sections of 5 microns thickness was prepared. They were considered in four different groups like A, B, C andD. Tissue sections in Group A were stained with H & E method where xylene was used as deparaffinizing agent. Theother three sections were stained with H & E where dish wash solution, diluted lemon water and coconut oil wereused as deparaffinising agent’s alternative to Xylene. Staining characteristics were compared with xylene andscoring was given. The total score of 3–5 was regarded as satisfactory for diagnosis and less than that isinsufficient for diagnosis.Statitistical Analysis: Chi square test was used.Results: Adequacy of staining characteristics such as nuclear, cytoplasm, uniformity, clarity and crispiness ofstaining for diagnosis was greater with dish wash solution followed by diluted lemon water, coconut oil andxylene.Conclusion: The Eco-Friendly deparaffinizing agents such as dish wash solution, diluted lemon water, and coconutoil can be used as alternatives to xylene

Synthesis and biological evaluation of new peroxo-bridged diosgenin derivatives / 中草药·英文版

Objective: In order to find lead compound with anti-HBV activity from peroxo-bridged diosgenin derivatives obtained with Eosin Y as the photosensitizer. Method: Eosin Y was used as the photosensitizer to activate the oxygen in the air to synthesize novel diosgenin derivatives with peroxo-bridge. The structures of synthesized compounds were identified by NMR and HR-MS. Their cytotoxicity and antihepatitis B activity were evaluated via MTS assay and ELISA method, respectively. Results: Six diosgenin derivatives were synthesized, three of which contained peroxo-bridge, and their structures were confirmed by spectroscopy. It showed that 5α,8α-peroxo-6-alkenyl-diosgenin (7) could suppress the production of HBsAg on transfected HepG2.2.15 cells at low-toxic concentration and the inhibition rate on HepG2.2.15 cells was 18.28% at 12.50 µg/mL, better than that of 3TC (7.30% at 12.50 µg/mL) and others. Conclusion: Due to its lower cytotoxicity and potential anti-hepatitis B activity, compound 7 could be developed as the promising candidate of anti-hepatitis B drug. It also indicated that the peroxo-bridged derivatives had potential biological values for developing clinical agents.