Therapeutic effects of proanthocyanidin and coenzyme Q10 on nitrogen mustard-induced ocular injury / Efeitos terapêuticos da proantocianidina e da coenzima q10 em lesão ocular induzida por mostarda de nitrogênio

ABSTRACT Purpose: Nitrogen mustard (NM) is a devastating casualty agent in chemical warfare. There is no effective antidote to treat NM-induced ocular injury. We aimed to assess the effects of proanthocyanidin (PAC) and coenzyme Q10 (CoQ10) on NM-induced ocular injury. Methods: Eighteen male rats were divided into the following 4 groups: NM, NM + PAC, NM + CoQ10, and control. The 3 NM groups received a single dose of NM (0.02 mg/μL) on the right eye to induce ocular injury. The control group received saline only. Thirty minutes after the application of NM, the NM + PAC group received PAC (100 mg/kg) via gastric gavage, while the NM + CoQ10 group received CoQ10 (10 mg/kg) via intraperitoneal injection. PAC and CoQ10 were administered once a day for 5 consecutive days. The rats were then sacrificed. Macroscopic images of the eyes were examined and eye tissues were collected for histology. Results: The treatment groups were compared to the control group with regard to both corneal opacity and lid injury scores. The findings were not significantly different for both the NM + PAC and NM + CoQ10 groups. In both the NM + PAC and NM + CoQ10 groups, the histological changes seen in the NM group demonstrated improvement. Conclusions: Our results indicate that PAC and CoQ10 treatments have therapeutic effects on NM-induced ocular injury in a rat model. PAC and CoQ10 may be novel options in patients with NM-induced ocular injury.

RESUMO Objetivo: A mostarda de nitrogênio (MN) é um agente de guerra química devastador. Não existe um antídoto eficaz para tratar lesões oculares induzidas por MN. Nosso objetivo foi avaliar os efeitos da proantocianidina (PAC) e da coenzima Q10 (CoQ10) na lesão ocular induzida por MN. Métodos: Dezoito ratos machos foram divididos em 4 grupos: MN, MN + PAC, MN + CoQ10 e Controle. Três grupos receberam uma dose única de MN (0,02 mg/μL) destilada no olho direito para gerar lesão ocular. Os animais do grupo controle receberam apenas solução salina. Trinta minutos após a aplicação de MN nos animais, o grupo MN + PAC recebeu PAC (100 mg/kg) por gavagem gástrica, enquanto a CoQ10 (10 mg/kg) foi administrada ao grupo MN + CoQ10 por meio de injeção intraperitoneal. A administração de PAC e de CoQ10 foi realizada uma vez por dia, durante 5 dias consecutivos. Os ratos foram, então, sacrificados. Imagens macroscópicas dos olhos foram examinadas e tecidos oculares foram coletados para histologia. Resultados: Os grupos de tratamento foram comparados ao grupo de controle quanto à opacidade da córnea e quanto aos escores de lesão da cobertura da córnea. Os resultados foram insignificantes para ambos os grupos. Ambos, o grupo MN+PAC e o grupo MN+CoQ10, apresentaram melhoras das alterações histológicas observadas no grupo MN. Conclusões: Nossos resultados indicam que os tratamentos com PAC e com CoQ10 têm efeitos terapêuticos sobre lesões oculares induzidas por MN em um modelo em ratos. A proantocianidina e a CoQ10 podem ser uma nova opção nesses casos.

Paraclinical findings in Iranian veterans exposed to sulfur mustard gas: A literature review / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>Sulfur mustard (SM) causes various systemic disturbances in human beings. This study aimed to assess paraclinical changes caused by exposure to SM gas in Iranian veterans during the war between Iraq and Iran.</p><p><b>METHODS</b>A literature review was carried out in international and national medical databases including ISI, Medline, Scopus, Iranmedex and Irandoc. Both Farsi and English literature were searched.</p><p><b>RESULTS</b>Search of the literature yielded 422 medical articles related to SM poisoning. Among them, 30 relevant articles were thoroughly reviewed. The most important reported complications were leukopenia, neutropenia, lymphopenia, eosinophilia, thrombocytopenia, increased bleeding time, positive C-reactive protein (CRP), rheumatoid factor (RF), antinuclear antibody (ANA), decreased T helper cells, natural killer cells, IL6, and IL8 levels, elevation of serum immunoglobulins, decreased levels of T3, T4 and cortisol, increased level of adrenocorticotropic hormone (ACTH), proteinuria, hematuria, and elevated liver enzymes. Also, there were some changes in chest assessments.</p><p><b>CONCLUSIONS</b>SM causes profound systemic complications in victims, even years after exposure. The paraclinical changes can be observed in hematology, immune system, biochemistry, hormonal profile and some imaging studies.</p>

Poisoning mechanism and protective drugs of sulfur mustard: An update / 第二军医大学学报

Sulfur mustard (SM) is a bifunctional alkylating agent that can react with multiple biochemical molecules such as DNA, RNA, proteins, and so on, and its alkylation with DNA is one of the major poisoning mechanisms. Presently the pathogenesis of SM included DNA alkylation, poly ADP-ribose polymerase (PARP) activation, oxidative stress, inflammation, activation of immunoregulation and proteolytic enzymes, etc. By now there have been no specific antidotes in clinical treatment, and all the existing drugs are mainly used for symptomatic treatment. The drugs used clinically and currently being under development include free radical scavengers, antioxidant agents, PARP inhibitors, anti-inflammatory drugs and protease inhibitors. This review summarized the advances in pathogenesis of SM and the corresponding protective drugs.

Pathologic changes of corneal lesions in rabbits after ocular exposure to mustard gas / 中华创伤杂志

Objective To observe the pathologic changes of corneal lesions in rabbits induced by mustard gas and changes of lymphocytes in the pathologic development so as to discuss the role of lymphocytes in the pathogenic process.Methods Thirty-five New Zealand white rabbits were randomly divided into experiment group and control group according to the random number table.In experiment group,rabbits were exposed to 0.2 ml of mustard gas liquid (400 μl/L) for a period of 4 minutes and were divided into 6 subgroups at 15 min,2 h,1 w,3 w,4 w,and 8 w after injury.Corneal tissue from each group was collected to detect changes of lymphocytes with aid of HE and immunohistochemical stainings.Results In experiment group,corneal epithelium was dropped totally,basement membrane was exposed,swelling was thickened,matrix layer was loosened and swelled,and collagen fibre was ranged loosely at 15 minutes and two hours ; corneal epithelium appeared multi-layer hyperplastic change in acute restoration period at 1 week; basal corneal epithelium cells appeared irregular column arrangement in chronic inflammation period at 3 and 4 weeks; around 10％ of toxic corneas had delayed reaction with partial corneal epithelium resloughed,fundus membrane appeared,edema thickened,and stroma collagens loosened and swelled at eight weeks.Immunohistochemical Envision staining showed positive expression of plymphomonocytes (CD20,CD45RO,and LCA) inside matrix layer at corneoscleral junction.Conclusions Ocular exposure to 400 μ/L mustard gas for a period of four minutes leads to direct and delayed lesions to cornea,which presents at one week and eight weeks respectively.The lymphocyte-mediated cellular and humoral immunity responses may be involved in the pathogenic process.

Therapeutic effect of equol on local skin wounds caused by mustard gas in rabbits and related mechanism / 第二军医大学学报

Objective To study the therapeutic effect of estrogenic equol (Eq) on mustard gas-caused skin wounds and the possible mechanism. Methods A rabbit skin wound model was established by contaminating the hip skin with mustard gas (0. 5 mg/cm2). The contaminated wound was treated externally with different concentrations (10, 50, and 100 μmol/L) of Eq for a week, and the changes of the wound were observed macroscopically. Wound specimens were collected for histopathological evaluation. Cell apoptosis and DNA cycle changes were analyzed by flow cytometry. Levels of IL-6 and TNF-α in the contaminated wound specimens were detected. Results The scores of erythema, edema, erosion and necrosis of the contaminated wounds in the treatment groups were significantly lower than those in the control group 120 h after mustard gas exposure (P<0. 05). The scores for changes of histopathological inflammation and necrosis were significantly lower in the 50 μtmol/L Eq treatment group than those in the control group 168 h after mustard gas exposure (P<0. 05). The results of flow cytometry showed that the proportion of apoptotic cells in the control group was significantly higher than those in the treatment groups 168 h after mustard gas exposure (P<0. 05), and the proportion of cells in DNA (S+G2-M) phase in the treatment groups was significantly higher than that in the control group (P<0. 05). The levels of IL-6 and TNF-α in the treatment groups were significantly lower than those in the control group 168 h after mustard gas exposure (P<0. 05). Conclusion Eq can promote healing of mustard gas-caused local skin damaged by inhibiting inflammatory response and cell apoptosis and enhancing cell proliferation.

Experiences Living with Fatigue in Iranian Veterans Chemically Injured by Sulfur Mustard Gas: A Phenomenological Study

PURPOSE: Fatigue affects the quality of life. Evidence shows that the phenomenon of fatigue is experienced differently depending on the type of disease and its consequences. The aim of the study was to explicate the meanings of the experience of living with fatigue in chemically injured veterans. METHODS: The hermeneutic phenomenology approach was used in this study, with an emphasis on Van Mennen's viewpoint and approach. According to Van Mennen, six overlapping dynamic activities are recommended to conduct a phenomenological study. During unstructured interviews, the participants were asked to describe their daily living experiences with fatigue. The participants were individuals who were chemically injured due to exposure to mustard gas. After examining every statement in the interview text, extractions of the meaning units, clustering, and themes were performed. RESULTS: The data explication was based on the third to sixth stages of Van Mennen's approach. The experience living with fatigue was classified into four essential themes: fatigue as a chronic condition, as an unstable and affected situation, as a physical condition of the entire individual, and as a mental condition of the entire individual. CONCLUSION: Due to unique social interactions and pathogenicity, victims of mustard gas experience fatigue differently than patients with other chronic diseases.

Effect of mustard gas at LD50 dose on reproductive system of rats / 第二军医大学学报

Objective: To evaluate the effect of mustard gas on reproductive system in rats. Methods: Forty male SD rats and 40 female rats were equally divided into mustard gas-treated group and control group. Animals in mustard gas-treated group were injected with 5 mg · kg body weight (LD50) of mustard gas, and those in control group received propylene glycol injection. Serum hormonal determinations (estriol, testosterone, luteinizing hormone, follicle-stimulating hormone) were completed by collecting ocular vein blood at 1 day, 3 days, 5 days, 7 days and 28 days after injection. The rats were then killed were anatomized. Microscope was used to observe pathological changes in germ cells (ovary or testes). Two female rats and 3 male rats died during 4th and 5th day after injection. Results: The changes of serum hormone and pathology nearly restored to the normal levels in the female rats after injection of mustard gas for a short period. Most of the spermatogenic cells were still necrotic and seminiferous tubules still had hollow cross 1 month after injection in male rats. The FSH levels in the sera of male rats were significangtly increased from (1.210 0±0. 354 6) mmol/L in control group to 0.475 0±0. 045 1) mmol/L 28 days after injection(P<0.01). Conclusion: Mustard gas causes slight injury of ovary in female rats and the damage can recover in a short period, but it can cause long-time and unreversable injury to testes in male rats.

Therapeutic effect of bone marrow transplantation on mustard-poisoned mice / 中国药理学与毒理学杂志

AIM To study the therapeutic effect of bone marrow transplantation on mustard gas-poisoned mice. METHODS Mustard-poisoned mice model was constructed by injected mustard gas 20 mg·kg~(-1) subcutaneously. At 4 h, 1 d and 2 d after poisoned by mustard gas, the suspended bone marrow cells (106 cells per time) were injected into mice respectively. After poisoned 3, 5, 7 and 10 d, the numbers of leukocyte and marrow nucleated cells were recorded periodically. At the same time, after poisoned 4 h and 1 d by mustard gas, mice in another group were injected granulocyte colony stimulating factor (G-CSF) 74 μg·kg~(-1) combined with transplantation and changes of the cell number were observed. RESULTS Compared with poisoned group without transplantion, leukocyte number in bone marrow transplanted group presented ascending trend, but there was no statistical significance until the 10 d after poisoned. However, the number of marrow nucleated cell increased significantly from the 5th day. After mice injected bone marrow cells combined with G-CSF, leukocyte increased markedly in earlier stage compared with transplantation only. CONCLUTION The bone marrow transplantation has significant effects on improvement of marrow hematogenic system inhibited by mustard gas.

Acute effects of sulphur mustard gas on the number of lymphocytes in the rat's spleen / Efectos agudos del gas mostaza sobre el número de linfocitos en el bazo de rata

Sulphur mustard (SM), commonly known as mustard gas is an alkylating agent that causes serious blisters upon contact with human skin. SM is frequently used as a chemical warfare agent. There is some evidence for sulfur mustard-induced lymph system effects in humans. Between 2000-2001, 42 male albino Wistar rats were used. After accommodation with environment, we divided rats to control, sham and experimental groups (2.5 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg and 40 mg/kg). Then we injected sulphur mustard oil in rat's intraperitoneal space. Then their spleens were removed for histological verification. Our results showed that significant difference in lymphocytes number in experimental groups after 24 hours. The number of lymphocytes in 5, 10, 20 and 40 mg/kg groups was increased and this increase in 40 mg/kg group was more than the other groups. We concluded that the number of lymphocytes increased due to exposure of mustard gas and there is a relationship between the increase of lymphocytes and dose of exposure.

El sulfuro de mostaza (SM), comúnmente conocido como gas mostaza, es un agente alquilante que causa graves ampollas en contacto con la piel humana. SM se utiliza con frecuencia como un agente de guerra química. Hay algunas evidencias que indican que el SM induce efectos en el sistema linfático en seres humanos. Entre los años 2000-2001, fueron utilizadas 42 ratas albinas Wistar macho. Después de la acomodación con el medio ambiente, las ratas se dividieron en grupos control, impostor y experimental (2,5 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg y 40 mg / kg). Luego se inyectó aceite de SM en el espacio intraperitoneal de las ratas. A continuación, sus bazos fueron removidos para la verificación histológica. Los resultados mostraron una diferencia significativa en el número de linfocitos en el grupo experimental después de 24 horas. El número de linfocitos en los grupos de 5, 10, 20 y 40 mg / kg fue mayor siendo este incremento en el grupo de 40 mg / kg más alto que en los otros grupos. Concluimos que el número de linfocitos aumenta debido a la exposición de gas mostaza existiendo una relación entre el aumento de linfocitos y la dosis de exposición.

Chromosomal aberration and micronucleus formation induced by sulfur mustard / 第三军医大学学报

Objective To study the effects of sulfur mustard on chromosomal aberration in Chinese hamster fibroblast cell line (CHL cells) and the micronucleus (MN) frequency in the bone marrow cells of mice. Methods CHL cells (with and without S9) treated with different concentrations of sulfur mustard for 24 h were harvested for the preparation of chromosome. Subsequently, the frequency and the type of chromosomal aberration were examined. The KM mice were sacrificed at 24 h after subcutaneous injection of sulfur mustard. The number of micronuclei-containing polychromatic erythrocytes (PCEs) was counted by means of bone marrow smear and Giemsa staining. Results Sulfur mustard might increase the chromosomal aberration frequency of CHL cell in a dose-dependent manner. S9 had no effects on the classificaiton and the frequency of chromosomal aberration. The types of chromosomal aberration included gap, break, deletion, polyploid, and so on. Compared with that in the control group, the MN ratio in every group increased significantly (P

Transcription and expression of endothelin-1 and endothelin A receptor in lung after mustard intoxication / 中国病理生理杂志

AIM: To investigate the role of endothelin-1 (ET-1), endothelin A receptor (ETAR) in pathophysiology of the lung after mustard intoxication. METHODS: The changes of transcription and expression of ET-1 and ETAR in the lung and their relationship with the tissue damage were studied with immunohistochemistry and RT-PCR. RESULTS: The level of ET-1 in the lung was significantly increased at 2 h, but decreased at 12 h after mustard intoxication. The transcription and expressin of ET-1 and ETAR were significantly increased at the early stage after intoxication. mRNA of endothelin-1 was increased, while mRNA of endothelin A receptor was down-regulated after (12 h.) CONCLUSION: ET-1 and ETAR in lung may play important roles in the pathophysiological process of mustard intoxication. [

THE INFLUENCE OF MUSTARD GAS AND OTHER CDEMICAL AGENTS TO THE KINDS OCEANIC LIVINGS / 第二军医大学学报

The samples of sea water and oceanic living things, including 8 kinds of animals and 21 plants were collected from Dinghai Qingdao and Yulin harbor in China and were experimented with mustard gas, lewisite, sarin and VX. The results showed that when fishes and crabs were in sea water of 5-35℃ which contaminated by mustard gas (50-750mg/l) for several minutes, they will appeared symptoms and died one hour later.The color of 12 kinds plants of Chlorophyta and Phaeophyta were changed just a few minutes after contacted with mustard gas and other agents.It also showed that effects of acid and alkali on plants of Phaeophyta have the same action as chemical agents as mentioned above.This method is simple and practical.Combining with some signs of chemical attack in war, it can be used for detection of gases, so it has some practical significance.

Effect of mustard gas at LD50 dose on reproductive system of rats / 第二军医大学学报

Objective:To evaluate the effect of mustard gas on reproductive system in rats.Methods:Forty male SD rats and 40 female rats were equally divided into mustard gas-treated group and control group.Animals in mustard gas-treated group were injected with 5 mg?kg body weight(LD_(50))of mustard gas,and those in control group received propylene glycol injection.Serum hormonal determinations(estriol,testosterone,luteinizing hormone,follicle-stimulating hormone)were completed by collecting ocular vein blood at 1 day,3 days,5 days,7 days and 28 days after injection.The rats were then killed were anatomized.Microscope was used to observe pathological changes in germ cells(ovary or testes).Two female rats and 3 male rats died during 4th and 5th day after injection.Results: The changes of serum hormone and pathology nearly restored to the normal levels in the female rats after injection of mustard gas for a short period.Most of the spermatogenic cells were still necrotic and seminiferous tubules still had hollow cross 1 month after injection in male rats.The FSH levels in the sera of male rats were significangtly increased from(1.2100?0.3546)mmol/L in control group to (1.4750?0.0451)mmol/L 28 days after injection(P

3-year follow up survey of 39 patients with skin lesion caused by mustard gas / 解放军医学杂志

Objective To investigate the remote effects of epidermal growth factor(EGF)and early removal of eschar combined with skin grafting for the treatment of skin lesion produced by mustard gas.Methods A 3-year follow-up survey was condueted in 39 patients with skin lesion produced by mustard gas.The condition of healing was observed and evaluated.Results Good epithelial healing with pigmentation was observed in all the surveyed patients,and only in a few of them showed local recurrence of wound.Itching and paresthesia in the wounded parts were the main complaints.The sequelae remained unpredictable.Conclusion The early use of epidermal growth factor and early removal of eschar followed by skin grafting for the treatment of skin lesion produced by mustard gas may achieve satisfactory therapeutic effect.

Morphological study of injuries to cells of peripheral blood and bone marrow as a result of mustard gas poisoning / 解放军医学杂志

Objective To study the morphological changes in peripheral blood cells and bone marrow cells of the patients suffering from mustard gas poisoning and their relationship with the degree of poisoning. Method The peripheral blood cells and marrow cells were examined morphologically after mustard gas poisoning. Results The total WBC count was reduced progressively in 44 patients, among them 10 patients showed granulocytopenia or agranulocytosis. Severe injury to eosinophils was seen also in the early period, and the percentage and absolute value of lymphocytes were significantly lowered too. However, platelets were not significantly influenced. The red cell count and hemoglobin level were elevated because of hemoconcentration. Blood marrow cells showed marked morphological changes. WBC became swollen with appearance of poisonous granules and vacuoles in their cytoplasm. Deformed lymphocytes (LY) could be found in the marrow cells of all the patients, mainly in the form of immature lymphocytes. It was shown that mustard gas produced serious damage to the hematopoitic cells of the bone marrow in the early period. Conclusions The examination of pheripheral blood and bone marrow cells is an important measure to determine the degree of mustard gas poisoning. The decrease in the absolute value of lymophocyte count and the time of its recovery, and degree of inhibition of hematopoiesis are closely related to the degree of mustard gas poisoning.

The analysis of eye injury and treatment in “8?4” mustard gas poisoning accident / 解放军医学杂志

Objective To investigate the incubation period, degree of injury, distribition of pathological changes, and therapeutic effect of eye injury in “8?4” mustard gas poisoning accident. Methods The relationships among contact pattern, contact time and incubation period and degree of injury were analyzed, and the distribition of pathological changes and therapeutic effect were assessed. Results An incubation period was found in eye injury after the exposure to mustard gas. It varied with the mode of contact and the time of exposure to the gas. The conjunctiva was the main site of injury, follwed by the eyelid, and then the cornea and the iris. Conclusions The length of the incubation period varied with the degree of injury and contact time and mode. Thorough washout and medicinal intervention should be instituted as early as possible.

Effect of basic fibroblast growth factor on healing of rabbit corneal wound caused by mustard gas / 第二军医大学学报

Objective: To study the effect of topical administration of basic fibroblast growth factor (bFGF) on healing of rabbit corneal wound caused by mustard gas. Methods: All the corneas of 8 New Zealand white rabbits were injured by liquid mustard at the concentration of 0.2%. One eye of each rabbit was treated with bFGF eye drops 6 times daily, the other eye with 0.25% chloromycetin eye drops as control. Computerized image analyzer was used to calculate the corneal epithelial healing rate. Results: The mean epithelial healing rate in bFGF-treated eyes was (1.276?0.152) mm 2/h, whereas in the control eyes it was (1.094?0.154) mm 2/h. The difference of 2 groups was statistically significant (P