Comparación Morfométrica de Esqueletos Transparentados de Cuatro Especies de Roedores Myomorfos / Morphometric Comparison of Transparent Skeletons of Four Species of Myomorphic Rodents

Las técnicas de doble tinción y transparentación se han usado desde 1897, pero su utilidad ha sido poco explorada en los estudios anatómicos de micromamíferos adultos. No obstante, la combinación de estas técnicas con el análisis alométrico mutivariado posibilita el estudio de esqueletos poscraneales articulados de tales grupos de micromamíferos como los roedores, los cuales son muy limitados ya que casi siempre se enfocan en los cráneos. En este estudio, analizamos y comparamos la morfometría del esqueleto de Neotomodon alstoni con la de Meriones unguiculatus, Phodopus campbelli y Rattus norvegicus. Usamos la técnica de doble tinción y transparentación para analizar las relaciones morfométricas entre estos roedores utilizando sesenta caracteres esqueléticos. Se encontró que tres especies comparten dos correlaciones comunes y compartieron el mismo tipo de crecimiento isométrico en una de ellas; además se encontraron similitudes aparentes entre los patrones de la morfometría de P campbelli con el patrón de osificación descrito para la especie relacionada Mesocricetus auratus. Las diferencias en el crecimiento alométrico pueden representar también diferencias en el ritmo de desarrollo de acuerdo con el tipo de historia de vida de cada especie. Aquí demostramos que tanto la técnica de preparación como el método de análisis morfométricos son herramientas poderosas pero simples, para realizar estudios anatómicos y morfológicos en el laboratorio. Nuestros resultados reflejan las condiciones del desarrollo ontogenético derivados delpropio patrón de heterocronía para cada especie, y además representan la historia evolutiva de este grupo analizado. Sin embargo, consideramos que es deseable más investigación.

SUMMARY: Clearing and staining techniques have been present since 1897, However, their use in anatomical studies of adult micromammals has been limited. When using such techniques in combination with allometric method, it is possible to study articulated skeletons of micromammals, instead of relying only on the skulls, which is important in morphologically complicated groups as the rodents. Research involving multivariate allometric analysis of postcranial skeleton of rodents has been limited and confined to specific items. In this study, we analyzed and compared the morphometry of the skeleton of Neotomodon alstoni with that of Meriones unguiculatus, Phodopus campbelli and Rattus norvegicus. We applied the double staining and clearing technique in order to determine the morphometric relation between these rodents using sixty skeletal characters. We found that three species share two common correlations and one isometric, with apparent similarities between the morphometry patterns of P campbelli with the ossification pattern described for the related species Mesocricetus auratus. The differences in allometric growth could represent differences in the development stages according to the type of life history for each species. In this analysis we confirmed that both the preparation technique and morphometric analysis method, are simple yet verifiable tools for anatomical and morphological studies. Our results reflect the conditions of ontogenetic development derived from the heterochrony pattern for each species, representing the evolutionary history for this group. Therefore, as this approach continues to be discussed, ongoing research is warranted.

Effect of Moringa olifera leaves on growth and gut microbiota of Nile tilapia (Oreochromis niloticus) / Efeito das folhas da Moringa olifera no crescimento e na microbiota intestinal da tilápia do Nilo (Oreochromis niloticus)

Abstract The study was conducted to evaluate the effect of Moringa olifera on the growth and gut health of Tilapia (Oreochromis niloticus). The feed having 30% crude protein was prepared as an experimental diet with 4%, 8% and 10% M. olifera leaf supplementation, respectively. The control diet was devoid of M. olifera leaves. The 10 weeks feeding trial was carried out on 60 fish in aquaria. Fish was fed @ 3% of body weight twice a day. Diet with the high level of inclusion of M. olifera leaves significantly increased the growth rate, Survival Rate (SR), Specific Growth Rate (SGR) and Feed Conversion Efficiency (FCE) in all treatment groups compared to the control group. Similarly, Feed Conversion Ratio (FCR) gradually decreased and found highly-significant. To check the gut health of the Tilapia, random samples were selected and dissected. Nutrient agar was used as culture media to check the growth of bacteria. Pour Plate Method was used for viable colonies count by colony counter. Through staining method, the different bacteria such as Escherichia coli, Salmonella, Shigella and Pseudomonas aeruginosa were identify abundantly in the intestine of control diet fish but less number present in treatment diets groups. These results showed that M. olifera leaves up to 10% of dietary protein can be used for Nile tilapia for significant growth and healthy gut microbiota of fish.

Resumo O estudo foi conduzido para avaliar o efeito da Moringa olifera no crescimento e saúde intestinal da tilápia (Oreochromis niloticus). A ração com 30% de proteína bruta foi preparada como dieta experimental com 4%, 8% e 10% de suplementação de folhas de M. olifera, respectivamente. A dieta controle foi desprovida de folhas de M. olifera. O ensaio de alimentação de 10 semanas foi realizado em 60 peixes em aquários. O peixe pesava 3% do peso corporal duas vezes ao dia. A dieta com alto nível de inclusão de folhas de M. olifera aumentou significativamente a taxa de crescimento, taxa de sobrevivência (SR), taxa de crescimento de sobrevivência (SGR) e eficiência de conversão alimentar (FCE) em todos os grupos de tratamento em comparação com o grupo de controle. Da mesma forma, a taxa de conversão de alimentação (FCR) diminuiu gradualmente e foi considerada altamente significativa. Para verificar a saúde intestinal da tilápia, amostras aleatórias foram selecionadas e dissecadas. O ágar nutriente foi usado como meio de cultura para verificar o crescimento das bactérias. O método da placa de Verter foi usado para a contagem de colônias viáveis ​​por contador de colônias. Através do método de coloração, diferentes como Escherichia coli, Salmonella, Shigella e Pseudomonas aeruginosa foram identificados abundantemente no intestino de peixes da dieta controle, mas em menor número nos grupos de dieta de tratamento. Esses resultados mostraram que M. olifera deixa até 10% da proteína dietética e pode ser usado para tilápia do Nilo para um crescimento significativo e microbiota intestinal saudável de peixes.

Effect of Moringa olifera leaves on growth and gut microbiota of Nile tilapia (Oreochromis niloticus)

Abstract The study was conducted to evaluate the effect of Moringa olifera on the growth and gut health of Tilapia (Oreochromis niloticus). The feed having 30% crude protein was prepared as an experimental diet with 4%, 8% and 10% M. olifera leaf supplementation, respectively. The control diet was devoid of M. olifera leaves. The 10 weeks feeding trial was carried out on 60 fish in aquaria. Fish was fed @ 3% of body weight twice a day. Diet with the high level of inclusion of M. olifera leaves significantly increased the growth rate, Survival Rate (SR), Specific Growth Rate (SGR) and Feed Conversion Efficiency (FCE) in all treatment groups compared to the control group. Similarly, Feed Conversion Ratio (FCR) gradually decreased and found highly-significant. To check the gut health of the Tilapia, random samples were selected and dissected. Nutrient agar was used as culture media to check the growth of bacteria. Pour Plate Method was used for viable colonies count by colony counter. Through staining method, the different bacteria such as Escherichia coli, Salmonella, Shigella and Pseudomonas aeruginosa were identify abundantly in the intestine of control diet fish but less number present in treatment diets groups. These results showed that M. olifera leaves up to 10% of dietary protein can be used for Nile tilapia for significant growth and healthy gut microbiota of fish.

Resumo O estudo foi conduzido para avaliar o efeito da Moringa olifera no crescimento e saúde intestinal da tilápia (Oreochromis niloticus). A ração com 30% de proteína bruta foi preparada como dieta experimental com 4%, 8% e 10% de suplementação de folhas de M. olifera, respectivamente. A dieta controle foi desprovida de folhas de M. olifera. O ensaio de alimentação de 10 semanas foi realizado em 60 peixes em aquários. O peixe pesava 3% do peso corporal duas vezes ao dia. A dieta com alto nível de inclusão de folhas de M. olifera aumentou significativamente a taxa de crescimento, taxa de sobrevivência (SR), taxa de crescimento de sobrevivência (SGR) e eficiência de conversão alimentar (FCE) em todos os grupos de tratamento em comparação com o grupo de controle. Da mesma forma, a taxa de conversão de alimentação (FCR) diminuiu gradualmente e foi considerada altamente significativa. Para verificar a saúde intestinal da tilápia, amostras aleatórias foram selecionadas e dissecadas. O ágar nutriente foi usado como meio de cultura para verificar o crescimento das bactérias. O método da placa de Verter foi usado para a contagem de colônias viáveis por contador de colônias. Através do método de coloração, diferentes como Escherichia coli, Salmonella, Shigella e Pseudomonas aeruginosa foram identificados abundantemente no intestino de peixes da dieta controle, mas em menor número nos grupos de dieta de tratamento. Esses resultados mostraram que M. olifera deixa até 10% da proteína dietética e pode ser usado para tilápia do Nilo para um crescimento significativo e microbiota intestinal saudável de peixes.

Prevalence of Taenia solium taenaisis in Kanchipuram district of Tamil Nadu (India): A cross-sectional study

Background: Taenia solium infections in humans include the infection by the adult tapeworm, these infections are of public health concern and are among the most important afflictions of humans who live in areas of poverty in the developing world and least developed countries. T. solium, a zoonotic disease, transmitted between pigs and humans and among humans, is common in developing countries. Aims and Objectives: The aim of the study was to estimate the detection rate of T. solium taeniasis among patients and random community screening with an indication of intestinal parasitic infection by routine stool examination. Materials and Methods: Stool samples were collected from the community and patients. Those who were willing, samples were screened for the cysts/ova/egg by direct microscopic examination by saline, iodine, concentration technique, and modified acid fast staining, were performed to differentiate species of T. solium and Taenia saginata. Results: Overall samples were 2030, out of which 870 stool samples were from community field screening 585 (28.81%) were positive. 1160 from tertiary care center, 668 (32.90%) were positive gave a total prevalence of intestinal parasitic infection of 61.72%. The prevalence of T. solium taeniasis was 194 (9.55%) out of which 92 (4.53%) were from community and 102 (5.02%) were from tertiary care center. Conclusion: The high prevalence of intestinal parasitic infestation might be due to the poor sanitary, contaminated water, and lack of education that is prevalent in the studied region as in other pockets in rural India. Our study showed the usefulness of the Ziehl-Neelsen modified acid-fast stain for identification of Taenia species.

Efficacy and safety of topical 2% rebamipide ophthalmic suspension in dry eye disease at tertiary care centre

Purpose: To evaluate the effect and side effects of topical 2% rebamipide ophthalmic suspension in dry eye disease. Method: This prospective randomized case control study included total 80 patients (40 cases and 40 controls) of dry eye. Symptoms were graded according to OSDI scoring system and specific tests for dry eye included Tear film breakup time (TBUT), Schirmer’s test, Fluorescein corneal staining (FCS), Rose Bengal staining) were performed. Case group received 2% rebamipide ophthalmic suspension four times daily and control group given carboxymethylcellulose 0.5% four times daily. The follow ups had done at two, six and twelve weeks. Results: The maximum numbers of patients were between 45-60 years. Patient with mild moderate and severe OSDI Score shows marked improvement. Mild TBUT score showed improvement but statistically not significant (P value-0.34). In moderate and severe TBUT Score statistically significant improvement (P value- 0.0001, 0.0001). In all grade FCS shows statistically significant improvement with p value-0.0001, 0.0001, and 0.028 respectively. Schirmer’s test score in all cases had shown improvement but statistically not significant and P value were 0.09, 0.07, and 0.07 respectively. In mild, moderate and severe Rose Bengal staining statistically significant improvement (P value -0.027, 0.0001, and 0.04) .The only side effect was dysgeusia (10% patients). Conclusion: Rebamipide 2% ophthalmic suspension showed significant improvement in symptoms and signs of dry eye. It able to modify epithelial cell function, improve tear stability, and suppress inflammation suggests that it may be a first drug of choice for severe dry eye disease.

Anticancer potential of Solanum lycopersicum L. extract in human lung epithelial cancer cells A549

The study aimed to reveal the phytochemical profile, free radical scavenging potential, and anticancer activity of Solanum lycopersicum L. leaf extract (SLLE). According to the study, SLLE contains plant secondary metabolites that are beneficial for health, like phenolics, flavonoids, ascorbic acid, alkaloids, and terpenoids. The SLLE has shown potential free radical scavenging potential in DPPH and ABTS free radical scavenging analysis and its EC50 values (concentration required to inhibit 50% of free radicals) were determined as 481.29 ± 33.82 and 527.56 ± 20.34 µg/mL, respectively. The SLLE has the ability to scavenge free radicals and could be used to treat illnesses brought on by oxidative stress. The anticancer activity of SLLE was assessed by MTT, LDH, micro-morphological, live/dead dual staining, and caspase-3 analysis. In the MTT assay, the IC50 value (concentration required to inhibit 50% of cell viability) of SLLE was determined as 190.41 ± 4.77 µg/mL. Furthermore, SLLE has shown potential anticancer activity by adversely affecting the plasma membrane integrity and escalating the caspase-3 levels. In the biomedical field, SLLE could be highly useful to treat cancer.

Construction of ATMT System for Clonostachys rosea and Its Conditional Optimization of Colonization into Glycyrrhiza uralensis Seeds / 中国实验方剂学杂志

ObjectiveAgrobacterium tumefaciens-mediated transformation （ATMT） of Clonostachys rosea， an endophytic fungus of Glycyrrhiza uralensis seeds， was established and optimized， and orthogonal test was designed to optimize the colonization conditions of C. rosea for G. uralensis seeds， so as to lay foundation for the development of biofertilizer and the breeding of high-quality G. uralensis. MethodThe conditions of ATMT were optimized from three aspects， including the concentration of acetosyringone， co-culture time and the concentration of conidia of recipient fungi. Then， high-quality transformants were selected. Orthogonal test was used to optimize the colonization conditions by taking co-culture temperature， co-culture time and spore concentration as factors and colonization rate as index. ResultWhen spore concentration was 1×107 cfu·mL-1， acetosyringone concentration was 150 μmol·L-1 and the co-culture time was 60 h， the transformation efficiency of C. rosea was the highest， which was 135 transformants per 1×107 recipient fungal spores. The accuracy and stability of the transformations were tested by cloning the marker gene green fluorescent protein （GFP） and β-glucuronidase （GUS） staining. When co-culture temperature was 25 ℃， co-culture time was 36 h and the spore concentration was 1×106 cfu·mL-1， the colonizing rate for C. rosea back dyeing into G. uralensis seeds by seed soaking method was the highest， which was 71.11%. ConclusionThis study successfully establishes stable and efficient technical systems not only of ATMT in C. rosea， but also of colonization of the transformants into G. uralensis seeds， which can lay a foundation for the development of biofertilizer of G. uralensis.

Establishment of a mouse model of acute systemic cold injury induced by hypothermia / 中华急诊医学杂志

Objective:To establish an animal model of acute systemic cold injury in mice.Methods:There were 98 C57BL/6 mice, half male and half female, with body weight of 22-27 g and age of 10 weeks. The mice were randomly divided into 7 groups ( n=14) according to the changes of anal temperature in cold environment, namely, group A (38.5 ± 1) ℃, group B (35 ± 1) ℃, group C (30 ± 1) ℃, group D (25 ± 1) ℃, group E (20 ± 1) ℃, group F (15 ± 1) ℃, and group G (10 ± 1) ℃, among which, group A was the blank control group, and the rest groups were the experimental group. The mice in the blank control group were placed in the normal environment (20 ± 5) ℃, and the mice in the experimental group were placed in the low temperature artificial climate box at - 20℃. The anal temperature of the mice was measured intermittently (as the core temperature), and the time required for the core temperature of the mice to drop to groups B, C, D, E, F and G was recorded. The righting reflex was used to evaluate the consciousness state, the action ability and the general state of each organ of mice were observed, and the blood routine and HE staining of each organ were detected. Results:The lower the core temperature of the experimental group, the longer the time required. The consciousness state, action ability, general state of organs, blood routine, and HE staining of organs in groups B, C, and D were basically the same as those in group A, and there was no acute systemic cold injury. Therefore, the blood routine, general observation of organs, and HE staining of organs in groups B, C, and D were no longer displayed compared with those in group A. Compared with group A, mice in group E began to suffer from disturbance of consciousness and action ability. With the decrease of core body temperature, the damage was aggravated, and mice in group G died. Compared with group A, the indices of blood routine test (WBC, RBC, HGB, PLT) of mice in group E began to decrease, and the univariate variance calculation showed that only WBC changes had statistical significance ( P<0.05). Compared with groups A and E, the indices of blood routine test (WBC, RBC, HGB, PLT) of mice in group F were further reduced, and the changes of each index in univariate variance calculation were statistically significant ( P<0.05). The general observation results showed that compared with group A, the lung, liver and spleen surfaces of mice in group E began to darken, and compared with groups A and E, the lung, liver, spleen, kidney and heart of mice in group F were further deepened and darkened, with irregular edges. HE staining results of various organs showed that compared with group A, the mice in group E began to have partial alveolar structure destruction and a small amount of inflammatory cell infiltration, the central vein of the liver was slightly congested, and the red and white pulp of the spleen were indistinct. Compared with groups A and E, the pathological structure damage of the lung, liver, spleen, kidney, heart and brain tissues of the mice in group F was further aggravated. Conclusions:Detection of consciousness state, action ability, general state of organs, blood routine and HE staining indices of organs in mice under low temperature can simulate the progress of clinical acute cold injury, and the animal model of acute systemic cold injury was successfully prepared.

Practical Value of Environment-friendly Sample Release Agent Combined with Ultrasound in the Preparation of Pathological Tissue Sections / 中山大学学报(医学科学版)

ObjectiveTo explore the practical value of environment-friendly sample release agent combined with ultrasound in the preparation of pathological tissue sections. MethodsFrom February 2013 to December 2022， 2 518 pathological specimens submitted by Foshan Municipal Hospital of Traditional Chinese Medicine were selected as the study objects. Two samples of the same specimen were randomly divided into two groups： the environment-friendly fast group， in which the pathological tissue sections were made by using the environment-friendly sample release agent combined with ultrasound； and the traditional group， in which formaldehyde， ethanol and xylene were used to make slices in the conventional way. The differences of hematoxylin （HE） staining effect， immunohistochemistry （IHC） staining effect and MDM2 gene detection result of atypical lipomatous tumor/highly differentiated liposarcoma （ALT/WDL） tissue sections between the two groups were compared. Results① The wax of the two groups' pathological tissues was dehydrated well and the tissue hardness was moderate. After HE staining， the sections of the two groups were intact， without cracks and tremor marks， and the contrast between nucleus and cytoplasm was appropriate， with good transparency， uniform staining， and no tissue loss. The excellent rate and score of HE staining in the environmental fast group were higher than those in the traditional group， but the difference was not statistically significant （χ2 = 3.125，P1 = 0.070；t = 0.965，P2 = 0.334）. ②After IHC staining of the two groups of sections， the positive location of the cells was accurate， the staining was specific and uniform， the staining intensity was moderate， the staining sensitivity was good， and there was no tissue loss. The excellent rate of IHC staining and the positive rate of IHC staining in the environmental fast group were lower than those in the traditional group， but the difference was not statistically significant （χ12 = 2.769，P1 = 0.092；χ22 = 0.800，P2 = 0.375）. ③The background and outline of the two groups of WDL tissue sections were clear， the staining was uniform， the cells were clear and visible， the nuclear boundary was clear， the hybridization signal was clear and bright under the background fluorescence， and there was no miscellaneous signal. The two groups of sections were hybridized successfully， and MDM2 showed positive amplification. The number of cells successfully hybridized in the environment-friendly fast group was lower than that in the traditional group， but the difference was not statistically significant （t = 1.414，P = 0.230）. ConclusionsThe tissue treatment method of using environment-friendly sample release agent combined with ultrasound can ensure the detection effect of HE staining， IHC staining and MDM2 gene detection of pathological tissue sections， and is more efficient and environment-friendly， suitable for promotion and use in hospitals at all levels.

Development of a flow cytometry method for detection of bovine multi-cytokines / 生物工程学报

This study aims to develop a method to detect bovine multi-cytokines based on flow cytometry. Previously we have prepared and screened monoclonal antibodies against bovine cytokines IFN-γ, IL-2, TNF-α, IP-10 and MCP-1. These bovine cytokine monoclonal antibodies were fluorescently labeled, and the combination of antibody and cell surface molecules were used to develop the method for detecting bovine multi-cytokines. Subsequently, the developed method was used to determine the cytokine expression profile of Mycobacterium bovis BCG infected bovine peripheral blood mononuclear cells in vitro, and evaluate the cytokine expression level of peripheral blood CD4+ T cells of tuberculosis-positive cattle. The bovine multi-cytokine flow cytometry detection method can effectively determine the cytokine expression of BCG-infected bovine peripheral blood T lymphocytes. Among them, the expression levels of IFN-γ, IL-2, and TNF-α continue to increase after 40 hours of infection, while the expression levels of IP-10 and MCP-1 decreased. The combined detection of IFN-γ, IL-2, and TNF-α on CD4+ T lymphocytes in peripheral blood of cattle can effectively distinguish tuberculosis-positive and tuberculosis-negative samples. This method may facilitate evaluating the level of cellular immune response after bovine pathogen infection and vaccine injection.

Effects of black tea tooth staining previously to 35% hydrogen peroxide bleaching

Aim: To determine if the artificial staining with black tea (BT) influences the enamel microhardness before in-office bleaching and if BT staining is necessary to evaluate the efficacy of bleaching with 35% hydrogen peroxide Methods: Enamel/dentin blocks were randomized into groups according to the staining protocol (n=5/group): (CO) control ­ maintained in artificial saliva solution (AS); (BT4) immersed in black tea solution for 4 h; (BT24) immersed in black tea solution for 24 h. After the staining protocols, all specimens were kept in AS for one week, followed by bleaching (three sessions of HP application for 40 min). Knoop surface microhardness (kgF/mm2) was determined at baseline (T0), after staining (T1), after 7 days of storage in AS (T2), and after bleaching (T3). The color (∆E00) and coordinate changes (∆L, ∆a, ∆b) were measured using a digital spectrophotometer at T0 and T3. Data were submitted to one-way (∆E00, ∆L, ∆a, ∆b) or two-way ANOVA repeated measures (kgF/mm2) and Tukey's test (a=5%). Results: The staining protocols (BT4 and BT24) promoted significantly lower microhardness (T1 and T2, p<0.05) than CO, whereas CO was the only group to maintain microhardness values over time. Bleaching promoted perceptible ∆E00 without a significant difference among the groups regardless of the staining protocol (p=0.122). CO and BT4 showed no differences in terms of ∆L and ∆a (p>0.05), but BT4 displayed a higher ∆b than CO. Conclusion:The artificial staining with BT negatively affected the enamel surface microhardness and was not essential to evaluate the efficacy of 35% hydrogen peroxide bleaching

Application value of new assistive technology in keratopathy-associated cataract surgery / 国际眼科杂志(Guoji Yanke Zazhi)

Keratopathy-associated cataract, that is, on the basis of corneal disease, and later the development of lens opacity, seriously damage visual quality. In order to avoid corneal transplantation for some patients, partial visual quality can be restored. A comprehensive and accurate evaluation of the effect of corneal opacity on visual function is of great improtance for determining cataract surgery alone. Due to the opacity of the cornea, the operation is very difficult and challenging. Therefore, it is of clinical value to develop and use new assistive technologies, including capsule staining, endoillumination, pupil dialation technology, femtosecond laser assisted technology, etc., avoiding problems such as limited visibility and decreased light flow caused by corneal opacity and facilitating cataract surgery. This article reviews progress of assistive technologies for keratopathy-associated cataract, hoping to guide clinical application.

Influencing factors of corneal epithelial punctate staining in patients wearing orthokeratology / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the influencing factors of punctate staining of corneal epithelium in patients wearing orthokeratology.METHOD: Data of 280 cases wearing orthokeratology were collected, including 185 cases in the group without corneal staining and 95 cases in the group with corneal staining. All patients were examined for general distant vision, intraocular pressure, slit lamp, fundus examination after mydriasis, comprehensive optometry, corneal topography, corneal thickness, corneal endothelium, axial length, Schirmer Ⅰtest(SⅠt)before wearing lenses, meibomian gland loss score before wearing lenses, eccentric distance, interleukin-2(IL-2)in tear, tumor necrosis factor-α(TNF-α)content analysis and so on. The influencing factors of corneal epithelium punctate staining were analyzed by univariate and multivariate Logistic regression.RESULTS: There were significant differences in preoperative diopter, preoperative meibomian gland deletion score, IL-2, TNF-α and lens sediment between the two groups(all P&#x003C;0.05). Logistic regression analysis showed that diopter before wearing lenses was a protective factor for corneal epithelium punctate staining. Before wearing lenses, the loss of meibomian gland score, IL-2, TNF-α and lens deposits were the risk factors of corneal epithelium punctate staining. In the detection of corneal epithelium punctate staining, the comprehensive advantage of lens deposits was obvious, the specificity of lens deposits was higher, and the sensitivity of IL-2 was the highest. CONCLUSIONS: Before wearing lenses, diopter is the protective factor of corneal epithelium punctate staining, and the loss of meibomian gland score, IL-2, TNF-α and lens deposits are the risk factors of corneal epithelium punctate staining.

Development and application of a method for determination of activity and vitality of Hansenula polymorpha by carboxyfluorescein diacetate-propidium iodide dua fluorescent staining / 中国生物制品学杂志

@#Objective To develop a method for determination of activity and viability of Hansenula polymorpha by dual fluorescent staining with carboxyfluorescein diacetate(CFDA)and propidium iodide(PI).Methods The time durations(5,10,20,40,60 and 120 min for CFDA,5,10,15,20 and 25 min for PI)and working solution concentrations(50,100,200and 300 μg for CFDA,2,10,20 and 30 μmol/L for PI)for the dual fluorescent staining were optimized by single factor test. Under the optimal condition,the H.polymorpha samples at theoretical survival rates of 0,25%,50%,75% and 100%were determined,of which the fluorescent intensity was observed under fluorescent microscope,and the gray value was analyzed by ImageJ software. The live and dead cells were counted,based on which the actual survival and death rates were calculated. Meanwhile,the relationships of actual gray value,actual survival rate and actual death rate to the corresponding theoretical values were analyzed. Activity and viability of three batches of cultured H.polymorpha were detected by CFDA-PI dual fluorescence staining.Results The optimal time durations for staining with CFDA and PI were 60 and 5 min,while the optimal working solution concentrations were 200 and 2 μmol/L,respectively. The actual gray value,actual survival rate and actual death rate of H.polymorpha samples at various theoretical survival rates were significantly correlated to the corresponding theoretical values(R~2=0. 998 3～0. 999 2,P < 0. 05). The CVs of activity and viability values in three detections of three batches of H.polymorpha culture were 3. 20%～4. 03% and 1. 10%～2. 27%, respectively.Conclusion The CFDA-PI dual fluorescent staining was successfully developed,which may be used for determination of activity and vitality of H.polymorpha.

Application value of endoscopic Nd-Fe-B magnet ring tracer technique to laparoscopy for colorectal lesions (with video) / 中华消化内镜杂志

Objective:To explore the clinical application value of preoperative colonoscopic marking by Nd-Fe-B magnet ring to assist laparoscopy.Methods:A total of 51 patients with colorectal tumor who underwent radical laparoscopy from January 2020 to October 2021 at the Department of Gastrointestinal Surgery, Guizhou Provincial People's Hospital were recruited. The patients were marked by Nd-Fe-B magnet ring under endoscopy one day before the operation, another magnet ring was sent into the abdominal cavity during the radical laparoscopy through cannula. The two magnet rings were attracted and clung to each other to orient the lesions. The basic information of patients, location of preoperative marks under endoscopy and laparoscopy conditions were recorded.Results:All 51 Nd-Fe-B magnet rings were successfully located to the position of colorectal tumor and fixed. According to the location of the lesions, there were 15 cases of transverse colon, 12 cases of descending colon, 19 cases of sigmoid colon, and 5 cases of upper rectal segment. According to the lesion type, there were 21 cases of colon cancer, 25 cases of polyp carcinomatosis, and 5 cases of laterally spreading tumors with partial carcinomatosis. There were 5 cases with positive margins after endoscopic mucosal resection and 1 case with positive margin after endoscopic submucosal dissection. All lesions were accurately located during the operation. The marking time was 4.1±1.2 min (3-6 min) before the operation and the localization time was 1.5±1.1 min (0.9-5.3 min) during the operation. All magnet rings were removed from the body by laparoscope. The mean distances between the tumor and the cutting edge of the proximal and distal intestinal segments were 5.5 cm and 6.3 cm, respectively. No complications such as colon mucosal injury, bleeding, intestinal perforation or local inflammatory reactions occurred.Conclusion:Nd-Fe-B magnet ring tracer technique for laparoscopic orientation is simple, fast, accurate and safe with no need for additional equipment or apparatus, which is worthy of clinical application.

Analysis of LMP specific T cell immune response and clinical factors in stage III-IVa nasopharyngeal carcinoma / 中华放射肿瘤学杂志

Objective:To explore the Epstein-Barr virus (EBV) latent infection membrane protein (LMP) 1 or LMP2 specific T cell immune response and clinical significance in stage III-IVa nasopharyngeal carcinoma (NPC), aiming to provide ideas and evidence for immunotherapy in NPC.Methods:Fifty-nine NPC patients admitted to the Affiliated Tumor Hospital of Xinjiang Medical University from February 2018 to October 2020 for primary treatment were collected. Peripheral blood monocytes (PBMCs) were stimulated by LMP antigen. Intracellular cytokine staining and flow cytometry were applied to study the expression levels of IL-2, IL-13, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) from CD4 + T and CD8 + T cells, and then analyzed in conjunction with clinical factors. Results:The positive rates of total PBMCs to LMP1 and LMP2 in NPC patients were different. The positive rate of LMP1 specific CD4 + T cells was statistically higher in stage T 3-T 4 NPC than that in stage T 1-T 2 (51.0% vs. 10.0%, P=0.042). There were also differences in the expression of cytokines between LMP1 and LMP2, CD4 +T cells and CD8 +T cells. Survival analysis showed the 2-year and 3-year overall survival (OS) rates were 91.5% and 88.2%, and the 2-year and 3-year progression-free survival (PFS) rates were 83.3% and 75.3%. Univariate analysis suggested that smoking history, male and LMP1 stimulated IL-13 positive expression in CD4 + T cells affected the disease progression ( P=0.026, 0.045 and 0.006); multivariate analysis showed LMP1 stimulated IL-13 positive expression in CD4 + T cells and smoking history were the independent prognostic factors affecting PFS ( P=0.017, 0.019). Conclusions:LMP1 and LMP2 generate specific T-cell immune response in PBMCs of NPC patients, with differential expression in two T-cell subsets. LMP1 and LMP2 specific T cell immune response is associated with primary tumor size and metastatic lymph node volume. LMP1 stimulated IL-13 positive expression in CD4 + T cells and smoking history affects the disease progression.

Assessment of Qualitative and Quantitative Changes in 5 days stored Platelet Concentrates in Bangabandhu Sheikh Mujib Medical University

Background: The fast-growing demand for platelet concentrates (PC) necessitates the storage of these blood products before transfusion. Platelets are prepared as concentrates from the whole blood or by plateletpheresis. Qualitative and quantitative assessment of these PCs is an important issue in transfusion medicine. To assess the qualitative, quantitative changes and bacteriological safety of 5 days of stored platelet concentrates (PC).Material & Methods:This prospective study was conducted at the department of Clinical Pathology in collaboration with the Department of Transfusion Medicine, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka from April 2008 to April 2009. A total of 65 healthy donors were included in the study as per the inclusion and exclusion criteria. Therefore, 65 platelet concentrates (bags/units) were prepared from the donors. Purposive sampling of the units was done. pH and platelet indices (PLT, MPV, PDW and P-LCR) were measured and Gram staining of PCs was performed on days 0 and 5. Statistical significant tests were done at a 95% confidence interval using the statistical package for social science (SPSS).Results:The mean (±SD) pH was 7.18±0.07 ranging from 7.0 to 7.3 during day 0. On day 5 the mean (±SD) pH was 6.77±0.11 and their range was from 6.5 to 7. The mean pH difference was statistically significant (p<0.05) between day 0 and day 5. The mean (±SD) PLT/unit was 70.56±15.56 x109/unit and it ranged from 38.01 to 110.6 x109/unit during day 0. On day 5 the mean (±SD) PLT/unit level was 68.46±15.52 x109/unit and it ranged from 36.82 to 107.2 x109/unit. The mean PLT/unit difference was statistically significant (p<0.05) between day 0 and day 5. The mean (±SD) MPV was 9.34±0.92 fl and it ranged from 7.5 to 11.5 fl during day 0. During day 5 the mean (±SD) MPV was 9.27±0.99 fl ranging from 7.0 to 11.2 fl. The mean (±SD) PDW was 10.07±1.61 fl and which ranged from 7.4 to 14.4 fl during day 0. During day 5 the mean (±SD) PDW was 10.72±1.71 fl ranging from 7.0 to 15.4 fl. The mean (±SD) PLCR was 18.28±5.67 % and it ranged from 8.0 to 32.5 % during day 0. During day 5 the mean (±SD) PLCR was 21.18±5.91 % and it ranged from 10.0 to 36.3 %. The mean PLT, PDW and PLCR differences were statistically significant (p<0.05) between day 0 and day 5 in the unpaired t-test, however, the mean MPV difference was not statistically significant (p<0.05) between day 0 and day 5. Gram staining of platelet concentrates on day 0 and day 5 found no bacteria.Conclusions:Storage-induced lesions take place in PCs when stored for 5 days in second-generation storage containers under the currently recommended conditions, but how far these changes are clinically relevant needs to be investigated.

Assessment of Qualitative and Quantitative Changes in 5 days stored Platelet Concentrates in a tertiary care Hospital of Bangladesh

Background: The fast growing demand for platelet concentrates (PC) necessitates the storage of these blood products prior to transfusion. Platelets are prepared as concentrates from the whole blood or by plateletpheresis. Qualitative and quantitative assessment of these PCs are an important issue in transfusion medicine. Aim of the study: To assess the qualitative, quantitative changes and bacteriological safety of 5 days stored platelet concentrates (PC).Material & Methods:This prospective study was conducted at the department of Clinical Pathology in collaboration with the department of Transfusion medicine, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka during April 2008 to April 2009. A total of 65 healthy donors were included for the study as per the inclusion and exclusion criteria. Therefore, 65 platelet concentrates (bags/units) were prepared from the donors. Purposive sampling of the units was done. pH and platelet indices (PLT, MPV, PDW and P-LCR) were measured and Gram staining of PCs were performed on day 0 and 5. Statistical significant tests were done at 95% confidence interval using statistical package for social science (SPSS).Results:The mean (±SD) pH was 7.18±0.07 ranging from 7.0 to 7.3 during day 0. During day 5 the mean (±SD) pH was 6.77±0.11 and their range was from 6.5 to 7. The mean pH difference was statistically significant (p<0.05) between day 0 and day 5. The mean (±SD) PLT/unit was 70.56±15.56 x109/unit and it ranged from 38.01 to 110.6 x109/unit during day 0. During day 5 the mean (±SD) PLT/unit level was 68.46±15.52 x109/unit and it ranged from 36.82 to 107.2 x109/unit. The mean PLT/unit difference was statistically significant (p<0.05) between day 0 and day 5. The mean (±SD) MPV was 9.34±0.92 fl and it ranged from 7.5 to 11.5 fl during day 0. During day 5 the mean (±SD) MPV was 9.27±0.99 fl ranging from 7.0 to 11.2 fl. The mean (±SD) PDW was 10.07±1.61 fl and which ranged from 7.4 to 14.4 fl during day 0. During day 5 the mean (±SD) PDW was 10.72±1.71 fl ranging from 7.0 to 15.4 fl. The mean (±SD) PLCR was 18.28±5.67 % and it ranged from 8.0 to 32.5 % during day 0. During day 5 the mean (±SD) PLCR was 21.18±5.91 % and it ranged from 10.0 to 36.3 %. The mean PLT, PDW and PLCR difference were statistically significant (p<0.05) between day 0 and day 5 in unpaired t-test, however the mean MPV difference was not statistically significant (p<0.05) between day 0 and day 5. Gram staining of platelet concentrates on day 0 and day 5 found no bacteria.Conclusions:Storage-induced lesions take place in PCs, when stored for 5 days in second generation storage containers under the currently recommended conditions, but how far these change are clinically relevant need to be investigated

Adventitial Collagen Cross-Linking by Glutaraldehyde Reinforcing Human Saphenous Vein - Implication for Coronary Artery Bypass Grafting

ABSTRACT Introduction: A weak venous wall is one of the major reasons contributing to vein graft failure after coronary artery bypass grafting (CABG). We investigated whether adventitial collagen cross-linking by glutaraldehyde reinforces venous wall, preserving the endothelium of veins during high-pressure distention. Methods: Human saphenous veins (SVs) were collected from 40 patients undergoing CABG, and adventitia cross-linking was performed with 0.3% glutaraldehyde for five minutes. The cross-linked SVs were accessed by biodegradation assay, immunofluorescent staining, and tensile test. Native SVs and cross-linked SVs from another 20 patients received the 200 mmHg pressure distention for two minutes. Pressure-induced injury of SVs were accessed by immunohistochemistry and electron microscopy. Results: Time to digestion was 97±13 minutes for native SVs and 720±0 minutes for cross-linked SVs (P<0.05). After adventitial cross-linking, the collagen I fibres of the vein remarkably presented with compact and nonporous arrangement. In the high-stretch region (stretch ratio 1.4-1.8), the Young's elastic modulus of stress-stretch ratio curve in cross-linked SVs was larger than that in native SVs (13.88 vs. 5.83, P<0.05). The cross-linked SVs had a lower extent of endothelial denudation without fibre fracture during high-pressure distension than native SVs. Comparing with the non-cross-linked SVs, the percentage of endothelial nitric oxide synthase staining length on the endothelium of cross-linked SVs was significantly preserved after high-pressure distension (85.2% vs. 64.7%, P<0.05). Conclusion: Adventitial collagen cross-linking by glutaraldehyde reinforced venous wall by increasing stiffness and decreasing extensibility of SVs and mitigated the endothelial damage under high-pressure distension.

MicroRNA-124a Protects the Myocardium Against Ischemia Reperfusion Injury Through Regulation of the Notch Signaling Pathway

Abstract Introduction: This study's objective is to investigate the effect of downregulation of micro ribonucleic acid (miR)-124a on myocardial injury after ischemia reperfusion (I/R) in rats. Methods: Sprague Dawley (SD) rats (n=20) were divided into four groups - sham, I/R, I/R+miR-124a antagomir (I/R+ant-miR-124a), and I/R+ant-normal control (NC). The pathomorphological and infarct size variance of injured myocardial tissues with IR were conducted with hematoxylin (HE) and triphenyltetrazolium chloride (TTC) staining. The expression levels of miR-124a, BAX, nuclear factor kappa B (NF-KB), Notch1, and Hes1 were examined by quantitative real-time polymerase chain reaction or Western blot in myocardium. The inflammatory cytokines interleukin (IL)-6, IL-1β, and tumor necrosis factor alpha (TNF-α) were detected by the enzyme-linked immunosorbent assay, as well as the activity of lactate dehydrogenase (LDH) and creatine kinase (CK) in serum by colorimetry. Results: The expression of miR-124a was increased in the I/R group. Compared with I/R and I/R+ant-NC groups, after downregulating miR-124a, the expression of IL-6, IL-1β, TNF-α, BAX, NF-KB, LDH, and CK were decreased, but the expression of Notch1 and Hes1 were increased. In HE staining, myocardial tissue edema, red blood cell exudation, and myocardial fiber arrangement disorder were accompanied by inflammatory cell infiltration and local necrosis in the I/R group. However, the pathological injury of myocardial tissue was alleviated after downregulating miR-124a. Additionally, TTC results showed that the myocardial infarction area was decreased in the I/R+ant-miR-124a group. Conclusion: Downregulation of miR-124a expression through Notch pathway can significantly reduce myocardial damage after 24 hours of I/R in SD rats. Therefore, miR-124a may become a potential therapeutic target for I/R injury.