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犬磷酸铵镁尿石症的诊疗及其中草药添加剂的研制
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摘要
犬尿石症属营养代谢性疾病,发病率占犬泌尿系统疾病的18%左右,近几年有逐渐增高趋势。尿石症的发病原因复杂,根据其分布位置分为肾结石、输尿管结石、膀胱结石、尿道结石,其临床病理因结石的大小、形状以及个体和品种差异而有所不同。据晶体和基质组成,主要有磷酸铵镁结石、草酸钙结石、磷酸钙结石、胱氨酸结石、硅石结石、尿酸铵结石和其他嘌呤结石或复合型结石。主要根据患犬的具体情况,结石的大小、形状,有无梗阻、感染、积水,结石的复发趋势等,选择合适的治疗方法。据报道,目前在北美犬的尿石症的发病率为0.54%,中国农业大学动物医院统计为1.50%,南京地区统计的尿石症的发病率0.58%一1.31%。据美国明尼苏达州结石定量分析中心1981—1997年对所有结石样本成分分析统计,犬磷酸铵镁结石占样品总数的49.6%(38285/77191),其中仅1997年就占45%(6923/15259)。因此,对临床上发病率最高的犬磷酸铵镁结石的研究有着极其重要的意义。
     本研究对南京地区临床上犬尿石症的发病情况进行了调查研究;对犬磷酸铵镁尿石症临床病例的诊疗进行了系统研究;研制出中草药添加剂,进行了临床犬磷酸铵镁尿石症的预防和治疗试验,结果证明对犬磷酸铵镁尿石症病例能起到很好的溶解,软化和排石作用,同时在防止手术后复发方面也有较好的效果。
     试验1南京地区犬尿石症的发病情况调查
     通过对2009年10月至2010年8月南京地区5家有代表性的动物医院临床犬尿石症的门诊186病例(经X线、B超和实验室检查,初步诊断为尿石症)的结石样本进行收集,并对成分进行离子鉴定,确定结石类型。调查样本病例检查结果显示各结石的发病率为:磷酸铵镁成分占51.08%(95/186)、草酸钙占24.19%(45/186)、磷酸钙占3.76%(7/186)、尿酸及尿酸盐占10.75%(20/186)、胱氨酸占1.08%(2/186)、复合型结石占8.06%(15/186)、混合性结石占1.08%(2/186)。患尿石症的发病犬年龄结构、品种特征、发病部位、季节特点及营养结构对结石的成分及发生部位有一定的关系。
     试验2犬磷酸铵镁尿石症的临床诊疗研究
     通过对2009-2010年公安部南京警犬研究所犬病研究中心医院收治的88例犬磷酸铵镁结石病例进行临床发病、治疗情况研究分析,所调查的患犬中以2-6岁的青年犬居多,约占收治尿石症病例的60%;我们对病例的基本情况统计表明,小型犬磷酸铵镁结石的发病率较高,为70.45%(62/88);母犬发病率为63.64%(56/88),远高于公犬发病率36.36%(32/88)。临床诊疗表明,通过对病犬的临床症状、X线和B超检查可初步诊断为犬尿石症,并能判知结石的部位;对取出的结石样本的成分分析可以确定结石的类型;通过营养结构调查分析,80%以上的病犬摄入过多动物内脏和瘦精肉从而导致尿液中的磷酸根离子、镁离子、铵离子增高,当达到一定浓度时,不溶性的磷酸铵镁就会饱和析出沉淀,从而引发结石。
     试验3预防和治疗犬尿石症的中草药添加剂的安全性研究
     根据中医理论组方,结合犬尿结石的发病机理,研制出纯中草药添加剂,主要成分包括:三棱、莪术、鸡内金、金钱草、黄芪、怀牛膝、核桃仁等。急性毒性试验中,中草药添加剂经小鼠口服的1日最大量56.25g/kg,相当于一头工作犬(30kg)用量的281倍(含量为1%时为281倍)、宠物犬(5kg)用量的225倍(含量为1%时为225倍),没有出现副作用和不良反应。从亚急性毒性试验结果可以看出,中草药添加剂可使大白鼠的红细胞、白细胞、血红蛋白的数量有所增加,体重平均日增重0.3225g,说明该中草药添加剂具有增强机体免疫力和促进动物生长的作用。本试验结果表明该中草药添加剂是安全、合理、有效的。
     试验4中草药添加剂预防犬磷酸铵镁尿石症术后结石复发的临床研究
     试验所选病例均为2007-2008年公安部南京警犬研究所犬病研究中心医院收治的门诊患犬,通过临床检查确诊为膀胱结石的小型犬,所取结石成分分析为磷酸铵镁结石,并于术后行X线B超检查,无结石。所选病例随机分为治疗组15例,对照组15例。试验组15例中,雄犬11例,雌犬4例;对照组15例中,雄犬10例,雌犬5例。所有犬尿石症手术后常规治疗;另试验组:加服中草药添加剂胶囊,1次/d,每次1粒,连续90天。经B超综合检测分析,术前膀胱壁厚度治疗组为3.82士1.01mm,对照组为3.79士0.96mm,两者无明显差异(P>0.05);试验1年后治疗组为2.95士0.41mm,对照组为3.40士0.52mm,两者有显著性差异(P<0.05);试验1.5年后治疗组为2.63±0.56mm,对照组为3.12士0.48mm,两者有显著性差异(P<0.05)。30天后镜检每个视野下的晶体数(分为无晶体、1-5个晶体、6个以上晶体),试验组无晶体为60%(9/15),1-5个晶体为40%(6/15);对照组无晶体为0,1-5个晶体为20%(3/15),6个以上晶体为80%(12/15);两组间治疗前后显微镜检查疗效分析,均有明显差异(P<0.05)。1年、1.5年内,治疗组的复发率均为0;对照组复发率分别为7.14%、20%;相同的时间段,治疗组的复发率显著低于对照组(P<0.05)。经上述数据的对比可充分证实中草药添加剂用于临床预防膀胱尿结石上有显著的疗效。
     试验5中草药添加剂治疗临床犬磷酸铵镁尿石症效果研究
     试验所选病例均为2008-2009年公安部南京警犬研究所犬病研究中心医院收治的门诊患犬,均为首次发病,临床上未发生尿闭的小型犬,且确诊为犬磷酸铵镁结石。所选病例随机分为治疗组30例,对照组15例。治疗组30例中,雄犬13例,雌犬17例,结石平均直径0.71±0.15cm。对照组15例中,雄犬6例,雌犬9例,结石平均直径0.65±0.14cm。所有试验犬常规治疗,治疗组加服中草药添加剂胶囊,1次/d,每次2粒,连续30天。经X光透视、B超疗效评定标准评定,试验30天后治疗组30例患犬检查发现治愈14例,显效8例,有效6例,无效2例,总有效率为93.33%。对照组15例患犬检查发现有1例治愈,显效1例,有效3例,无效10例,总有效率为33.33%。治疗组30天后临床症状、体征减轻或消失的有效率为88.89%,肉眼血尿有效率为100%,镜下血尿有效率为86.21%;对照组30天后临床症状、体征减轻或消失的有效率为23.08%,肉眼血尿有效率为1000%,镜下血尿有效率为42.86%。30天后镜检每个视野下晶体数,治疗组无晶体为46.67%(14/30),1-5个晶体为53.33%(16/30),6个以上晶体为0;对照组无晶体为0,1-5个晶体为33.33%(5/15),6个以上晶体为66.67%(10/15)。试验前治疗组与对照组各项指标无统计学意义(P>0.05);治疗组与对照组30天后总有效率、临床症状、体征及镜检疗效有显著性差异(P<0.05)。
Canine urolithiasis is the nutrition metabolic disease, and the incidence is about18%of the urinary system diseases, and increasing gradually in recent years. The cause of urolithiasis is complicated, which disease is divided into kidney stones, ureter stone, bladder calculi, urethra calculi according to the distribution position, and the clinical pathology is different because of the size, shape, individual and variety varies. According to crystal and matrix composition, urolithiasis is mainly divided into strurite calculi, calcium oxalate stone, calcium phosphate stones, cystine stones, silica stones, uric acid, and other ammoniumcalculicalculi or purine compound stone. We choose the suit treatment according to the calculi size, shape, with or without obstruction, infection, hydrops, the recurrence trend of calculi, and so on. It has been reported that, the incidence of urolithiasis in North America canine is0.54%, statistically for1.50%by animal hospital of China agricultural university, and the incidence of urolithiasis is0.58%-1.31%in nanjing area. The American Minnesota State calculi analysis center analysis all the stones samples from1981to1997year, founding that the calculi carbonated-apatite accounted for49.6%of the total sample number (38285/77191), and only in1997year is45%(6923/15259). Thus, the study of the clinical highest incidence of canine carbonated-apatite calculi is very important.
     This study is to reaserach the prevalence situation of clinical canine urolithiasis in nanjing region, and the system research of diagnosis and treatment in carbonated-apatite urolithiasis, developed the herbal medicine additive, and carried the prevention and treatment trial of the canine clinical canine carbonated-apatite urolithiasis, the experimental results proved that, the herbal medicine additive have very good dissolution, soften and row calculi effects of canine carbonated-apatite urolithiasis cases, and also have good effect of preventing recurrence after surgery.
     Experiment1Investigation of the prevalence situation of canine urolithiasis in Nanjing area
     Through collecting the calculi samples in five representative animal hospital clinical canine urolithiasis of outpatient service186cases (by X-ray, B ultrasonic and laboratory, a primary diagnosis is urolithiasis) from the October2009to August2010in Nanjing, identifing the ion of the ingredients, and determine the type of stone. The results showed that, phosphate magnesium accounted for51.08%(95/186), calcium oxalate accounted for24.19%(45/186), calcium phosphate accounted for3.76%(7/186), uric acid and uric acid salt accounted for10.75%(20/186), cystine accounted for1.08%(2/186), compound calculi accounted for8.06%(15/186), mixed stones accounted for1.08%(2/186). The age structure, breed characteristics, pathogenic sites, seasonal characteristic and nutrition structure of the disease canine have a certain relationship with calculi composition and the happening part.
     Experiment2Research of diagnosis and treatment of canine struvite urolithiasis
     We research the clinical morbidity and therapeutic condition of88cases of canine struvite urolithiasis, which were collected from2009to2010year in canine disease research center hospital of Nanjing police canine institute, and the age of2-6years young canine were in the majority, accounting for about60%of the cases; the statistics of the basic situation of case showed that, struvite stones of small canines had the higher incidence, it was70.45%(62/88); The incidence of bitch was63.64%(56/88), and far higher than male canines, whose incidence was36.36%(32/88). Clinical diagnosis and treatment showed that, according to canine clinical symptoms, X-ray and B ultrasonic examination, we can primary diagnosis and know the parts of the stone; and we can determine the type of calculi through analysising the component of stones samples; through the investigation and analysis of the nutrition structure, we found that, more than80%of the sickness canine intaked excessive entrails and thin essence, leading to increase phosphoric acid root ion, magnesium ions and ammonium ions in the urine, when the ions reach a certain concentration, the insoluble magnesium ammonium phosphate will be saturated and precipitate, then causing stone.
     Experiment3Safety evaluation of the herbal medicine additive on prevention and treatment
     According to TCM theory composing prescriptions and combining with the pathogenesis of canine urinary stones, we developed a pure Chinese herbal medicine additive, whose main ingredients including:Sanleng, Ezhu, Galli stomachichum corium, Loosestrife, Astragalus, achyranthis root, walnut kernel and so on. In acute toxicity trial, the maximum oral of herbal medicine additive in mice was56.25g/kg per day, equaling to281times (content was1%for281times) of the amount in working canine (30kg),225times (content was1%for225times) of the amount in pet canine (5kg), and we have not found the side effects and adverse reaction. In subacute toxicity trial, we found that, herbal medicine additive can increase the number of erythrocyte, leukocyte and hemoglobin, and the average daily gain weight was0.3225g, these results showed that, the herbal medicine additive can enhance immunity and promote the growth of animals. The experiment results showed that the Chinese herbal medicine feed additive is safe, reasonable and effective.
     Experiment4Clinical research of herbal medicine additive on prevention recurrence of stones after canine struvite urolithiasis surgery
     Test cases canine were selected from2007to2008year in canine disease research center hospital of Nanjing police canine institute, and through the clinical tests, the stone was diagnosed as bladder calculi of small canine, the calculi component was struvite stone, and carrying the X line and B ultrasonic examination, founding no stone. The selected cases randomly were divided into treatment group and control group,15cases respectively. In treatment group, the male dog was11cases, and the female dog was4cases; in control group, the male dog was10cases, female dog was5cases. After surgery, all urolithiasis canines had the conventional treatment; in experiment group:herbal medicine additive capsule added,1time/d, every time1grain, for90straight days. Through B-ultrasonography testing analysis, before surgery bladder wall thickness was3.82±1.01mm in treatment group, and was3.79±0.96mm in control group, no obvious difference (P>0.05) was found between the two group; after1year treatment, bladder wall thickness was2.95±0.41mm in treatment group, and was3.40±0.52mm in control group, significant difference (P<0.05) was found; after1.5years treatment, bladder wall thickness was2.63±0.56mm in treatment group, and was3.12±0.48mm in control group, significant difference (P<0.05) was found. After30days, obversing the crystal number in each field under microscopic (1-5crystal as no crystal, more than six as crystal), we found that, no crystal was60%(9/15) and1-5crystal was40%(6/15) in treatment group; no crystal was0,1-5crystal was20%(3/15), more than six crystal was80%(12/15) in control group; before and after treatment, efficacy analysis of examination under the microscope had significant difference (P<0.05) between the two groups. Within1year,1.5years, recurrence of the treatment group was0; the recurrence in control group were7.14%and20%; The same period, the recurrence of treatment group was significantly lower than control group (P<0.05). These results can fully confirmed that, herbal medicine additives have significant efficacy on preventing bladder urinary stones.
     Experiment5Effects of herbal medicine additive on canine struvite urolithiasis
     Test cases were selected from2008to2009year in canine disease research center hospital of Nanjing police canine institute, all the puppies was the first time to affect struvite urolithiasis, and have not urine closed in small dogs, and the diagnosis was struvite bladder calculi. The selected cases were randomly divided into treatment group of30cases and control group of15cases. In treatment group, the male dog was13cases, the female dog was17cases, and the average diameter of stone was0.71±0.15cm. In control group, the male dog was6cases, the female dog was9cases, and the average diameter of stone was0.65±0.14cm. All experiment canines had the conventional treatment; herbal medicine additive capsule was added in treatment group,1time/d, every time2grain, for30straight days. Through the effect evaluation standard evaluation by X-ray and B ultrasonic curative, after30treatment days, in treatment group,14cases were cured,8cases were markedly improved,6cases were effective and2cases were invalid, the total effective rate was93.33%; in control group,1case was cured,1case was marked effect,3cases were effective,10cases were invalid, the total effective rate was33.33%. After30days, the effective rate of clinical symptoms, signs relief or disappeare was88.89%, the effective rate of gross hematuria was100%, and the effective rate of hematuria was86.21%under mirror; the effective rate of clinical symptoms, signs relief or disappeare was23.08%, the effective rate of gross hematuria was100%, and the effective rate of hematuria was42.86%under mirror. After30days, obversing the crystal number in each field under microscopic, no crystal was46.67%(14/30),1-5crystal was53.33%(16/30) and more than six crystal was0in treatment group; no crystal was0,1-5crystal was33.33%(5/15) and more than six crystal was66.67%(10/15) in control group. Before experiment, each index had no statistically significant (P>0.05) between treatment and control group; after30experiment days, the total effective rate, clinical symptoms, signs and efficacy of microscopic examination had significant difference (P<0.05) between treatment and control group.
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