Origin and developmental patterns of lactase and other glycosidases in sheep amniotic and allantoic fluid.

Intestinal lactase activity (with its associated cellobiase, 4-methylumbelliferyl-beta-galactosidase and -beta-glucosidase activities) was used as a specific intestinal marker enzyme to study the release of protein and enzymes of intestinal origin in sheep amniotic fluid during gestation. In amniotic fluid, intestinal lactase activity peaked at 66--85 days of gestation and then decreased with gestation. This enzyme activity was very low or absent in allantoic fluid throughout gestation suggesting that there is no important transfer of amniotic fluid lactase towards the allantoic cavity. Maltase and 4-methylumbelliferyl-alpha-glucosidase showed no statistically significant variation with gestation in both amniotic and allantoic fluid whereas alpha-galactosidase and N-acetyl-beta-hexosaminidase which were first higher in allantoic than in amniotic fluid increased in amniotic fluid to reach allantoic fluid levels near term. Such patterns are consistent with the suggestion that the fetal urine is a source of alpha-galactosidase and N-acety-beta-hexosaminidase activities and that sheep urine is first accumulated in the allantoic sac via the urachus up to 86--90 days of gestation and thereafter passes more and more into the amniotic sac.     

Adenocarcinoma with a neuroendocrine component arising in the urachus. A case report

The clinical, cytologic, histologic and ultrastructural findings in a mixed adenocarcinoma and neuroendocrine carcinoma of the urinary bladder urachus, an extremely rare tumor only recently described, are presented for a 31-year-old woman who died of widespread metastatic disease six months following the initial diagnosis and treatment. Cytologic study of voided urine and bladder washings disclosed the presence of malignant cells with the features of a small cell carcinoma; retrospectively, scarce adenocarcinoma cells were also identified in those specimens. Histologic study of resection specimens, including the use of special stains and electron microscopy, confirmed the presence of a small cell component, consistent with the poor prognosis in this case. Image analysis measurements of the malignant cells suggested a high proliferation rate.     

Isoelectric profiles of human erythropoietin are different in serum and urine

By adding a step of immunoaffinity to the method we had previously developed for analysing erythropoietin (EPO) in urine, we were able to study the isoelectric profiles of this hormone in human serum samples. This method was sensitive enough to investigate samples presenting physiological levels of this hormone. Comparison with the corresponding profiles in urine showed that natural EPO was systematically more acidic in urine. The acidification process, which was not patent in the non-human primate Cynomolgus macaque, clearly also affected recombinant EPO when injected into humans. This process was unrelated to any enzymatic activity in urine since the incubation of natural or recombinant EPO in urine induced no transformation of their isoelectric profiles. The nature and mechanism of the structural modifications occurring during the renal handling of this hormone remain to be investigated.     

En Bloc Robot-assisted Laparoscopic Partial Cystectomy,Urachal Resection,and Pelvic Lymphadenectomy for Urachal Adenocarcinoma

Primary adenocarcinomas of the bladder and urachus are extremely rare, accounting for 0.5% to 2.0% of all bladder malignancies. During fetal development, the urachus develops into the median umbilical ligament that stretches from the umbilicus to the bladder. Adenocarcinoma accounts for 90% of all cases of urachal carcinoma. There is no consensus regarding the management of urachal carcinoma. Although the preferred treatment is wide local excision with partial or radical cystectomy, bladder-sparing management is increasing. We report a case of robot-assisted laparoscopic partial cystectomy with en bloc resection of the urachus and bilateral pelvic lymphadenectomy for urachal carcinoma. The robot-assisted laparoscopic approach allowed us to minimize surgical morbidity, postoperative pain, and convalescent time while maintaining the oncologic principle of wide local excision.Key words: Urachal adenocarcinoma, Robotic partial cystectomy, Pelvic lymphadenectomyPrimary adenocarcinomas of the bladder and urachus are extremely rare, accounting for 0.5% to 2.0% of all bladder malignancies.¹ The urachus is a musculofibrous band that extends from the dome of the bladder to the umbilicus. During fetal development, the urachus develops into the median umbilical ligament that stretches from the umbilicus to the bladder. Urachal carcinoma stems from the epithelium of the remnant of this structure, and adenocarcinoma accounts for 90% of all cases.² Historically, patients tend to have a poor prognosis, with 5-year survival rates of 11% to 61%.³ Patients with urachal carcinoma most commonly present with dysuria, hematuria, abdominal pain, or umbilical discharge.There is no consensus regarding the management of urachal carcinoma. Although the preferred treatment is wide local excision with partial or radical cystectomy, ^4–6 bladder-sparing management is increasing because the published reports do not clearly support a survival advantage with more radical extirpation.^7,8We report a case of robot-assisted laparoscopic partial cystectomy with en bloc resection of the urachus and bilateral pelvic lymphadenectomy for urachal carcinoma. The robot-assisted laparoscopic approach allowed us to minimize surgical morbidity, postoperative pain, and convalescent time while maintaining the oncologic principle of wide local excision.     

Prp-c and Prp-Sc at the fetal-maternal interface

Scrapie is a naturally occurring prion (PrP) disease causing a fatal neurodegenerative disorder in sheep and goats. Previous studies suggest that scrapie is transmitted naturally through exposure to the scrapie agent in wasted placentas of infected ewes. This study determined the distribution and biochemical properties of PrP cellular (PrP-C) and the distribution of PrP scrapie (PrP-Sc) in reproductive, placental, and selected fetal tissues and fetal fluids in sheep. Glycosylated, N-terminally truncated, proteinase K-sensitive PrP-C with apparent molecular masses of 23-37 kDa was present in reproductive, placental, and fetal tissues and fetal fluids. PrP-C was low or undetectable in intercotyledonary chorioallantois, amnion, urachus, amniotic fluid, and fetal urine. In pregnant ewes, cotyledonary chorioallantois, allantoic fluid, and caruncular endometrium contained higher levels of PrP-C than did intercaruncular endometrium, myometrium, oviduct, ovary, fetal bladder, or fetal kidney. Caruncular endometrial PrP-C was up-regulated during pregnancy. Despite the wide distribution of PrP-C in reproductive, placental, and selected fetal tissues and fetal fluid, PrP-Sc was detected only in caruncular endometrium and cotyledonary chorioallantois of pregnant scrapie-infected ewes. The embryo/fetus may not be exposed to scrapie in utero because it is separated physically from PrP-positive allantois and chorioallantois by PrP-negative amnion.     

A viable calf with trisomy 22

Karyotyping of eight malformed calves resulted in the detection of trisomy 22 in one calf. This calf had a hernia umbilicalis, a urachus fistula, and slight brachygnathia inferior. This trisomy was not associated with lethality.     

Function curve of the membranes that regulate amniotic fluid volume in sheep

Seven singleton 120-day fetal lambs were prepared with a shunt from the lung to the gastric end of the esophagus, a bladder catheter, and multiple amniotic fluid and vascular catheters. The urachus was ligated. Beginning 7 days later, amniotic fluid volumes were determined by drainage, followed by replacement with 1 liter of lactated Ringer (LR) solution. Urine flow into the amnion was measured continuously. In 14 of 27 experiments, amniotic fluid volumes were determined again 2 days after the inflow into the amnion had consisted of urine only and in 13 experiments after the inflow of urine had been supplemented by an intraamniotic infusion of LR solution. Intramembranous absorption was calculated from the inflows and the changes in volume between the beginning and end of each experiment. The relations between absorption rate and amniotic fluid volume, the "function curves," were highly individual. Urine production during the infusion of LR solution did not decrease, fetal plasma renin activity decreased (P < 0.001), and amniotic fluid volume increased by 140% [SE (27%), P < 0.005], but the increase in the amniochorionic absorption rate of 411% [SE (48%), P < 0.001] was greater (P < 0.005) than the increase in volume. Each of the seven fetuses was proven capable of an average intramembranous absorption rate that exceeded 4.5 liters of amniotic fluid per day. During the infusion of LR solution, the increase in the rate of absorption matched the rate of infusion (both in ml/h), with a regression coefficient of 0.75 (P < 0.001). Thus, even for large amniotic fluid volumes, volume is not limited by the absorptive capacity of the amniochorion, and, at least in these preparations, the position of the function curve and not the natural rate of inflow was the major determinant of resting amniotic fluid volume.     

Multiple vascular and gastric cannulation of swine for studies of gastrointestinal, liver, and peripheral tissue metabolism.

The implantation of multiple chronic indwelling vascular cannulas and of a gastric cannula into swine was described. Technics for housing, care, and handling of the animals pre- and post-surgically were established to reduce the incidence of infection and to allow collection of urine and feces in metabolic studies. This regimen has been used in 20 gilts to study absorption, tissue distribution, metabolism, and excretion of nutrients. On the average, the cannulas remained patent for 4 wk.     

Absorption of amniotic fluid by amniochorion in sheep

Swallowing of amniotic fluid and lung fluid inflow were eliminated in 10 chronically instrumented fetuses. The urachus was ligated, and fetal was urine drained to the outside. At the beginning and the end of 21 experiments of 66 +/- 5 (SE) h duration, all amniotic fluid was temporarily drained to the outside for volume measurement and sampling. Amniotic fluid osmolalities and oncotic pressures were experimentally controlled. Amniochorionic absorption of amniotic fluid depended strongly on the osmolality difference between amniotic fluid and fetal plasma (P < 0.001), but at zero osmolality difference there still was a mean absorption rate of 23.8 +/- 4.7 (SE) ml/h (P < 0.001). Absorption was unaffected by the protein concentration difference between amniotic fluid and fetal plasma, but infused bovine albumin in the amniotic fluid was absorbed at a rate of 1.8 8 +/- 0.4 g/h (P < 0.001), corresponding to a volume flow of fluid of 33.8 8 +/- 6.1 ml/h (P < 0.001). Fluid absorption in the amniochorion is driven in part by crystalloid osmotic pressure, but about 25 ml/h is absorbed by a path that is permeable to protein. That path has the physiological characteristics of lymphatic drainage, although no anatomic basis is known to exist for a lymphatic system in the amniochorion.     

水稻分子育种技术专利分析

通过Thomson Innovation（TI）专利平台数据库中相关专利的检索,利用专利计量学的方法,从专利申请数量、受理国家、申请机构、技术生命周期以及研发热点等方面对水稻分子育种技术国际发展态势进行分析。结果表明,水稻分子育种技术目前整体呈快速发展趋势,并已趋于成熟。其中,美国的申请量最多,是全球最受重视的技术市场。大型跨国公司不仅是全球最主要的专利申请主体,而且在我国的专利申请活动也较为活跃。我国在水稻分子育种技术领域专利数量较多,并且具有一定优势,相关专利申请主体为科研机构和大学。对专利文献内容的深入分析表明,重要农艺性状的基因挖掘和转基因技术是分子育种技术领域当前的研发热点。     

在我国失效的美国心血管疾病治疗药物的PCT 专利申请数据挖掘

目的：从没有在中国得到保护的美国专利申请或专利中寻找心血管疾病治疗药物研发的思路。方法：对申请日从2005 年1 月1 日至2014 年12 月31 日的美国心血管系统疾病治疗药物PCT 申请进入中国国家阶段后失效的专利申请及专利数据进行整序和分析,依据其在美国本土的法律状态确定其技术含金量。结果：未得到中国专利保护的美国心血管系统疾病治疗药物PCT 申请共174 件,其中170 件申请了美国优先权或进入了美国国家阶段。结论：170 件美国心血管系统疾病治疗药物PCT 申请在美国申请优先权,大多数因美国的优先权临时申请过期而放弃。14 件在美国本土的优先权申请失效,其所含信息量应该相当于公开发表的论文的信息量,23 件在美国本土优先权授权的专利技术具有一定的技术含金量,目前在中国已经进入公知公用领域,经过市场价值评估后可以无偿使用。     

Characteristics of genetic material with reference to patent documents

Characteristics of genetic material: plasmids, vectors, recombinant DNA and DNA fragments were, studied from 235 patent documents. These documents were from the patent offices of the Federal, Republic of Germany (38), the German Democratic Republic (15), the European Patent Office (53), Finland (43), Great Britain (65), the Soviet Union (3), and the United States of America (18). Attention was paid also to the terminology used in the patent documents. According to this study the way in which the genetic material was characterized in patent documents of different countries was quite variable, and the terminology used was sometimes confusing.     

Experimental onchocerciasis in chimpanzees. Antibody response and antigen recognition after primary infection with Onchocerca volvulus.

Nine of 18 chimpanzees inoculated with 250 infective third-stage larvae (L3) each developed patent (i.e., positive for microfilariae) Onchocerca volvulus infection. Four of 6 infected chimpanzees that received 200 micrograms/kg ivermectin at 28 days postinfection (pi) became patent, whereas, when ivermectin was given concurrently with L3 challenge only 1 of 6 infected animals developed patent infection. The antibody response to O. volvulus adult worm-derived antigens (OvAg) showed clear differences between patent and nonpatent chimpanzees. Three months pi, all sera detected several OvAg in the range of M(r) 35-120 k. Sera collected 6 mo pi from later patent animals recognized increasing numbers of OvAg, especially in the lower MW range of M(r) 13 to 33 k. Beginning 10 months pi Onchocerca-antigens of M(r) 21, 24, 26, and 28 k were detected only by patent chimpanzee's sera. The antibody response in nonpatent chimpanzees consistently recognized fewer OvAg, most of which were limited to the higher M(r) range (35-120 k). The reactivity of sera from infected chimpanzees to a low molecular weight fraction (LMW) of total OvAg doubled within 6 months pi, and increased continuously in patent animals from 13 until 30 months pi. Serological reactivity of nonpatent animals to LMW-OvAg remained low. The titers of circulating IgG directed against total OvAg increased in all infected chimpanzees, and continued to rise with patency. In nonpatent chimpanzees the antibody production gradually returned to preinfection values. Total and OvAg-specific IgE increased in patent and nonpatent chimpanzees. Also, during prepatency the granulocyte and antibody-mediated in vitro killing of microfilariae of O. volvulus increased in subsequently patent chimpanzees. The in vitro immobilization of L3 remained low.     

全球人冠状病毒抗体领域研发现状与发展趋势

抗体在疾病的诊断、治疗和预防方面发挥着重要作用。随着2019新型冠状病毒(SARS-CoV-2)感染引起的肺炎不断传播,如何研发针对该病毒的抗体迫在眉睫。基于IncoPat数据库的专利信息和Cortellis数据库的药物信息,采用定量分析与定性调研相结合的方法,从申请趋势、技术分布、国家/地区分布、机构分布以及市场现状等维度对人冠状病毒抗体领域的专利进行态势分析。结果表明美国是专利技术拥有量最大的国家,中国是专利保护的重点国家。中国科学院等3家国内机构进入全球前十位。人冠状病毒抗体产品研发主要以SARS和MERS为主,部分MERS抗体已进入临床阶段。研究结果为2019冠状病毒病(COVID-19)的相关抗体研发提供数据参考与决策支撑。     

心脏磁共振在不明原因脑卒中病因筛查中的应用

目的探讨心脏磁共振成像技术在不明原因缺血性脑卒中患者病因筛查中的应用价值。方法选取2014年9月~2015年2月在我院诊治的不明原因缺血性脑卒中患者53例为研究对象,均行心脏MRI和经胸超声心动图检查,并对检查出卵圆孔未闭(PFO)的患者进行分析。结果 53例不明原因缺血性脑卒中患者中,心脏MRI检查诊断合并PFO为27例(50.94%),经胸超声动图(TTE)检查诊断合并PFO为7例(13.21%),差异有统计学意义(P0.001)。结论心脏MRI对PFO的检出率较高,可作为不明原因缺血性脑卒中患者病因筛查的有效手段。     

专利微生物菌种的保藏与发放统计分析

对WIPO公开的"国际保藏机构2001～2010年专利微生物保藏与发放"数据进行了统计分析。结果表明:全世界的专利菌种保藏量与发放量处于快速增长中;美国是专利菌种保藏与发放量最多的国家;近年中国的专利菌种年保藏量实现三连冠,但专利菌种的发放率处于极低水平。     

中成药乙醚提取液对Raji细胞的EPSTEIN-BARR病毒早期抗原的诱导

应用Raji细胞系统研究了350种中成药,发现保赤散、小儿脐风散,铁娃丹,舟车丸,鹭鸶咯丸,时疫止泻丸、清心滚痰丸和八宝坤顺丸等8种中成药的乙醚提取液,对Raji细胞的EB病毒早期抗原(EA)有诱导作用,并讨论了这些中成药与肿瘤发生的关系。     

Evidence for a continual exchange of 5'-nucleotidase between the cell surface and cytoplasmic membranes in cultured rat fibroblasts

Approximately 40% of the 5'-nucleotidase activity in cultured rat embryo fibroblasts was patent, as judged by enzymatic assays comparing the activity of intact cells with detergent-solubilized cells. The patent activity was inhibited when cells were incubated with anti-5'-nucleotidase serum at 2 degrees C, whereas latent activity (calculated as the difference between total and patent activity) was not. Latent activity was inhibited by antibody when the antiserum was added directly to detergent-solubilized cells or when cells were cultured in the presence of antiserum for several hours. Patent activity was inhibited by antibody, and cells were returned to culture in antibody-free medium; after 12 hr, 30% of the total activity was expressed in intact cells and 60% of the anti-5'-nucleotidase, assayed by the binding of sheep antirabbit antibodies to intact cells, was lost from the cell surface, indicating an exchange of 5'-nucleotidase between the latent and patent compartments. Cytochemical studies showed that the patent activity was located on the cell surface and that latent activity was present in cytoplasmic vacuoles and vesicles, and in the Golgi complex. Over 30% of the anti-5'-nucleotidase internalized during 6 hr in culture returned to the cell surface after a further 9 hr, indicating a continual exchange of the enzyme between the cell surface and cytoplasmic membranes.     

The effect of ambient temperature on infrared thermographic images of joints in the distal forelimbs of healthy racehorses

The aim of the study was to describe the dependence on ambient temperature of distal joint temperature at the forelimbs of racehorses. The study also investigated the influence of differing ambient temperatures on the temperature difference between joints: this was measured ipsilaterally (i.e. between the carpal and fetlock joints along each forelimb) and contralaterally (i.e. between the same joints of the left and right forelimbs). Sixty-four healthy racehorses were monitored over 10 months. At each session, three thermographic images were taken of the dorsal, lateral and medial aspects of the distal forelimbs. Temperature measurements were made from regions of interest (ROIs) covering the carpal and fetlock joints. There was a strong correlation between ambient temperature and absolute joint temperature at all ROIs. The study also observed a moderate correlation between ambient temperature and the ipsilateral temperature differences between joints when measured from the medial and lateral aspects. No significant correlation was noted when measured dorsally. The mean contralateral temperature differences between joints were all close to 0 °C. The data support previous reports that the temperature distribution between the forelimbs of the healthy equine is generally symmetric, although some horses differ markedly from the average findings.