Preparation of 20 (S)-protopanaxadiol PLGA Nanoparticles

[Objectives]To prepare 20(S)-protopanaxadiol PLGA nanoparticles(20(S)-PPD-PLGA-NPs).[Methods]20(S)-PPD-PLGA-NPs were prepared by emulsion solvent evaporation method,and the optimal formulation was screened by Box-Behnken experiment with particle size and drug loading as the indicators through single factor experiment,and the drug release in vitro was carried out.[Results]The average diameter of the nanoparticles was(119.60±2.29)nm and the polydispersity index was(0.12±0.02),the size was uniform.The encapsulation efficiency and drug loading of protopanaxadiol were(87.99±1.29)%and(14.86±0.25)%,respectively.[Conclusions]The 20(S)-PPD-PLGA-NPs were successfully prepared by emulsion solvent evaporation method,and the 20(S)-PPD-PLGA-NPs had good stability,to lay a foundation for the study of 20(S)-PPD-PLGA-NPs in vitro and in vivo.

牛支原体疫苗研究进展

牛支原体(M.bovis)是牛的重要病原体,能够引起牛的乳腺炎、关节炎和肺炎等疾病。牛支原体已在全世界传播,对全球范围内的养牛业产生了重大不利影响。目前,由于牛支原体的耐药性有增强的趋势,同时缺乏有效的疫苗和治疗方法,这严重阻碍了牛支原体感染的控制。预防、控制甚至清除牛支原体最好的方法是使用有效的疫苗。虽然目前我国还没有获得新兽药证书的牛支原体疫苗,但相关的研究已取得了一定的进展。本文根据多年来牛支原体疫苗研究的相关资料,从牛支原体感染与免疫机制、灭活疫苗、弱毒活疫苗、基因工程疫苗四个方面进行综述,以期为今后牛支原体疫苗的研制提供思路及参考。

Preliminary exploration of animal models of congenital choledochal cysts

BACKGROUND Various animal models have been used to explore the pathogenesis of choledochal cysts(CCs),but with little convincing results.Current surgical techniques can achieve satisfactory outcomes for treatment of CCs.Consequently,recent studies have focused more on clinical issues rather than basic research.Therefore,we need appropriate animal models to further basic research.AIM To establish an appropriate animal model that may contribute to the investigation of the pathogenesis of CCs.METHODS Eighty-four specific pathogen-free female Sprague-Dawley rats were randomly allocated to a surgical group,sham surgical group,or control group.A rat model of CC was established by partial ligation of the bile duct.The reliability of the model was confirmed by measurements of serum biochemical indices,morpho-logy of common bile ducts of the rats as well as molecular biology experiments in rat and human tissues.RESULTS Dilation classified as mild(diameter,≥1 mm to<3 mm),moderate(≥3 mm to<10 mm),and severe(≥10 mm)was observed in 17,17,and 2 rats in the surgical group,respectively,while no dilation was observed in the control and sham surgical groups.Serum levels of alanine aminotransferase,aspartate aminotrans-ferase,total bilirubin,direct bilirubin,and total bile acids were significantly elevated in the surgical group as compared to the control group 7 d after surgery,while direct bilirubin,total bilirubin,and gamma-glutamyltransferase were further increased 14 d after surgery.Most of the biochemical indices gradually decreased to normal ranges 28 d after surgery.The protein expression trend of signal transducer and activator of transcription 3 in rat model was consistent with the human CC tissues.CONCLUSION The model of partial ligation of the bile duct of juvenile rats could morphologically simulate the cystic or fusiform CC,which may contribute to investigating the pathogenesis of CC.

Ecological Networks of Allometric Growth,Feeding Behavior and Gut Microbiota in Chinese Alligator Hatchlings

Chinese alligator hatchlings in captivity are fragile and have a high mortality rate within first year.The body mass gain of reptile animals is closely related to their feeding behavior and gut microbiota.This study analyzed the intrinsic factors influencing the allometric growth of Chinese alligator hatchlings based on their body mass gain,feeding behavior,and gut microbiota.This information would enhance the health management of Chinese alligator hatchlings.There was a significant correlation between the total distance moved,the average number of conspecifics nearby,and body mass gain.Chinese alligator hatchlings with a greater growth rate showed greater activity and more independent behavior during feeding than those with a lower growth rate.Moreover,after feeding started,some functions of the gut microbiota showed significant relationships with growth rate and feeding activity.Chinese alligator hatchlings with a greater growth rate showed greater levels of heme biosynthesis than those with a lower growth rate,and feeding activity was inhibited by long-chain fatty acid biosynthesis.These results elucidate the relationships between health,feeding behavior,and the gut microbiota of Chinese alligator hatchlings.Understanding the intrinsic factors of their health and feeding behavior can improve the health management of Chinese alligator hatchlings in captivity for conservation.

乌头汤治疗膝关节骨性关节炎作用机制研究进展

膝关节骨性关节炎(KOA)是中老年人常见的骨关节退行性疾病,严重影响患者的生活质量。乌头汤出自《金匮要略》,具有温经散寒、除湿,兼通达经络、止痹除痛的功效,是治疗KOA的常用方,尤其对寒湿痹阻型KOA疗效显著。现代药理研究表明,乌头汤通过调节炎性介质释放、降低一氧化氮含量、改善血液流变学指标等方式达到镇痛、延缓关节退变、改善微循环障碍等目的,以缓解KOA临床症状。下一步可从药物活性成分、药理作用等方面挖掘乌头汤治疗KOA的具体作用机制,同时进行药理毒理学研究,确保临床用药安全。

猪链球菌9型菌株的分离鉴定

为了确诊四川省某猪场1头25日龄左右的后腿跛行并急性死亡仔猪的病因,并制定对猪群其他猪只的预防和治疗方案。本试验剖检死亡仔猪,取脾脏、肝脏、肺脏组织和血液进行细菌分离培养,革兰染色镜检,PCR分型鉴定和药敏试验。结果显示,分离培养的细菌革兰染色为阳性;分离菌株经PCR检测为猪链球菌9型;药敏试验确定该分离菌株对氨苄西林、环丙沙星、氟苯尼考、青霉素、多西环素、头孢氨苄、氧氟沙星、头孢拉定、头孢呋辛和氯霉素敏感。结果表明,该猪场仔猪为感染猪链球菌9型引起的急性死亡,通过头孢类等药物可以有效的治疗发病仔猪。

Habitat Suitability and Distribution Pattern Response to Global Climate Change in a Widespread Species,the Asiatic Toad(Bufo gargarizans)

The distribution and diversity of the species are closely related to the global climate.As the most widely distributed species of Bufonidae in China,the study of the distribution pattern and habitat suitability of the Asiatic toad under climate change can help us understand the reply pattern of Bufonidae habitat to climate change.Here,combined with the Maxent model and GIS technology,the effects of climate change on the distribution pattern and habitat suitability of the Asiatic toad were comprehensively analyzed.The results show that the rainfall during the wettest season(Bio16)and the mean temperature of the driest season(Bio9)have a considerable impact on the distribution of the Asiatic toad.In the next 30 to 50 years,across the overall spacial scale of the Chinese mainland,the habitat of the Asiatic toad will be primarily in the eastern part of China and less in south part,while its distribution area will expand to the midwest and northwest parts of China.Overall,the area in which it can be distributed will be reduced and suitable habitat will shift to some regions of higher latitude and elevation.In a word,we systematically analyzed the changes of the distribution pattern and habitat suitability of the Asiatic toad with climate change,and we aim to provide data on how climatic variation may impact amphibians.

Data analysis guidelines for single‑cell RNA‑seq in biomedical studies and clinical applications

The application of single-cell RNA sequencing(scRNA-seq)in biomedical research has advanced our understanding of the pathogenesis of disease and provided valuable insights into new diagnostic and therapeutic strategies.With the expansion of capacity for high-throughput scRNA-seq,including clinical samples,the analysis of these huge volumes of data has become a daunting prospect for researchers entering this field.Here,we review the workflow for typical scRNA-seq data analysis,covering raw data processing and quality control,basic data analysis applicable for almost all scRNA-seq data sets,and advanced data analysis that should be tailored to specific scientific questions.While summarizing the current methods for each analysis step,we also provide an online repository of software and wrapped-up scripts to support the implementation.Recommendations and caveats are pointed out for some specific analysis tasks and approaches.We hope this resource will be helpful to researchers engaging with scRNA-seq,in particular for emerging clinical applications.

Host Genetic Background Impacts Microbiome Composition in Newborn Alligator

Genetic factors play a key role in determination of the structure of the cloacal flora for newborn Chinese Alligators.We collected the cloacal microbiomes for 24 newborn Chinese Alligators from three different genetic backgrounds for 16S gene amplicon sequencing.The number of cloacal flora for the Chinese Alligators from different groups was comparable but differed structurally.There were variations in proportions of floral compositions at the phylum and family levels;however,the main difference was at the genus level.There were two significant differences in richness and evenness among the three groups.Non-metric multidimensional scaling NMDS analysis revealed that the 24 samples could be clearly divided into three categories based on their genetic backgrounds(stress=0.0244).Thus,we postulated that newborn Chinese Alligators with different genetic backgrounds have different immune strengths,which affects individual responses to environmental microorganisms.In summary,newborn Chinese Alligators from different genetic backgrounds exhibit variations in cloacal microbiome.

利用关键点检测算法的超声图像定点测量

超声图像的定点距离测量在临床医学上十分重要.由于超声图像的噪声大,边缘模糊,因此关键点自动定位在超声图像中很有挑战性.目前的关键点检测算法通常是针对单个关键点位置进行优化,难以在保证每个关键点检测精度的情况下得到准确的测量距离.为使超声图像中关键点的精度和两个关键点之间的距离更加精确,本文提出一种基于级联卷积神经网络的关键点检测算法,该方法采用两个卷积网络从粗略到精细的对关键点进行定位.首先利用第一个网络回归两个关键点的粗略位置,并将包含这两个关键点的小区域送入第二个网络.然后本文提出一种加入距离修正的损失函数,作为第二个网络的优化目标,在第一个网络输出结果的基础上定位最终的关键点位置.实验结果表明,本文提出的级联方法无论是相比传统的级联方式还是回归树方法,本文算法在超声图像的关键点定位上更为精准,并且在最终的距离测量精度上也有很大的提高,在评价标准下比传统级联方法检测精度上提升将近30%.

基于逻辑回归树耦合熵指数模型的滑坡易发性分区——以陕西省延安市吴起县滑坡为例

研究合适的县域滑坡易发性分区的方法,对于滑坡的防治有着非常重要的现实意义。鉴于此,基于陕西省延安市吴起县的717个滑坡样本,选取坡度、坡向、高程、平面曲率、剖面曲率、年平均降雨量、距道路的距离、距河流的距离、岩土体类型和NDVI作为影响因子,计算对应的熵指数,构建了基于熵指数的建模数据集。随后,基于建模数据集,耦合熵指数(IOE)和逻辑回归树模型(LMT),建立了IOE-LMT混合分类模型,并绘制了吴起县滑坡易发性分区图。利用多种统计学指标、ROC曲线下的面积(AUROC)和平均绝对误差(MAE)评价分区精度和模型的泛化性能。结果表明:IOE-LMT模型的泛化性能较强(AUROC=0.942),且滑坡易发性分区图的精度较高;研究区内滑坡易发于黄土沟道范围内,并且研究区北部的滑坡易发性明显高于南部。评价结果合理可靠,可为当地的滑坡防治和国土空间规划提供参考。

1981~2016年甘肃省降水量时空变异性的信息熵分析

降水是具有时空属性的随机变量,准确刻画其时空分布规律对认识气候变化具有重要意义。基于1981~2016年甘肃省73个气象站点逐日观测资料,采用信息熵理论定量分析降水量年、季、月尺度上的不确定性,并运用Kriging插值法研究降水变异性分布格局。结果表明,不同时间尺度降水量变异性存在差异,具体为年>冬>秋>春>夏>月;月尺度上2、3月降水量变异性较大,7、8月降水量变异性较小。甘肃省降水量变异性呈东南向西北递减分布,河西地区降水量变异性最强、陇中次之、甘南和陇东地区最小。降水量越少的时空域变异性越大。

四自由度测量系统光束漂移抑制方法与系统

在机床导轨几何误差测量中,空气折射率不均匀造成的光束漂移会降低激光测量系统的检测精度,测量距离越远,影响越大。基于此,该文提出一种波纹管和气泵均匀化空气折射率的方法,抑制光漂的发生。以四自由度激光测量系统为对象,首先分析光束漂移机理,确定影响空气折射率的主要因素为温度与压强,给出轴向最大允许温度与压强梯度;其次通过流体仿真选取合适的波纹管内径,最后搭建实验系统并在不同条件下进行实验测试。结果表明:利用该系统可使四自由度激光测量系统的光束稳定性提高50%以上。该方法的可行性和有效性得到了验证。

Efficacy of concurrent chemoradiotherapy plus adjuvant chemotherapy on advanced cervical cancer

Background and Objective: Concurrent chemoradiotherapy for cervical carcinoma develops rapidly and has become a common and standard therapy in recent years. Both the local control rate and survival rate of patients were increased and the risk of death fell by 30%-50%. This study aimed to explore the efficacy of concurrent chemoradiotherapy plus adjuvant chemotherapy on and the treatment compliance of the patients with advanced cervical squamous cell carcinoma. Methods: A total of 156 patients with stage IIa-IIIb cervical squamous cell carcinoma were randomly divided into the concurrent chemoradiotherapy group (experimental group) and radiotherapy group (control group). Intracavity and external beam radiation therapy were administered. At point A, 40-48 Gy were given by 10-12 fractions; at point B, 46-50 Gy were given by 23-25 fractions. In the same time, experimental group was treated by cisplatin (DDP, 40 mg) on day 1, repeated every week. Ten days after radiation therapy, TP regimen was administered as adjuvant chemotherapy. Results: For the experimental and control groups, the objective response rates were 88.61% and 75.32%, 1-year survival rates were 88.57% and 70.77%, 1-year local control rates were 81.43% and 64.62%, 3-year survival rates were 82.14% and 57.69%, and 3-year local control rates were 75.00% and 46.15%, with significant differences (P < 0.05). Quality of life of all patients were significantly improved after treatment (P< 0.05). Conclusion: Concurrent chemoradiotherapy plus adjuvant chemotherapy for advanced cervical cancer can improve short-term and long-term survival and local control rates of patients, improve the quality of life, and the toxicity can be tolerated.

A New Species of the Genus Tylototriton(Amphibia: Urodela: Salamandridae) from the Southern Dabie Mountains in Anhui Province

A new species of the genus Tylototriton is described,from Yuexi county,Anhui province,in the south of the Dabie Mountains.It is based on morphological and molecular analysis.The new species is identified as belonging to the Tylototriton asperrimus group and shares a number of similarities with T.wenxianensis,T.broadoridgus and T.dabienicus.The diagnostic characteristics of the new species are as follows： the head length is greater than the width of the head; bony ridges on the head are prominent and necked-in; the distal digit ends,ventral digits,peripheral area of the cloaca and the tail＇s lower edge are orange.The result from the molecular analysis of the genus Tylototriton（including the type specimen of the new species） based on three mitochondrial genes（ND1,ND2 and CYTB） indicated that the new species was close to T.wenxianensis,T.dabienicus,and T.broadoridgus,but formed an independent clade.This result was consistent with the morphological analysis above,which supports the theory that the population distributed in the south of the Dabie Mountains,namely in from Yuexi county,Anhui province,represented a distinct species,Tylototriton anhuiensis sp.nov.

Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation

Transient receptor potential melastatin 2(TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression can help repair cerebral injury, and if so what the mechanism underlying this process involves. This study investigated the protective effect of reducing TRPM2 expression on pheochromocytoma(PC12) cells injured by oxygen-glucose deprivation(OGD). PC12 cells were transfected with plasmid encoding TRPM2 shRNAS, then subjected to OGD by incubation in glucose-free medium under hypoxic conditions for 8 hours, after which the cells were allowed to reoxygenate for 24 hours. Apoptotic cells, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels were detected using flow cytometry. The relative expression of C-X-C motif chemokine ligand 2(CXCL2), NACHT, LRR, and PYD domain–containing protein 3(NALP3), and caspase-1 were detected using fluorescence-based quantitative reverse transcription-polymerase chain reaction and western blotting. The rates of apoptosis, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels in the TRPM2-shRNA + OGD group were lower than those observed in the OGD group. Taken together, these results suggest that TRPM2 knockdown reduces OGD-induced neuronal injury, potentially by inhibiting apoptosis and reducing oxidative stress levels, mitochondrial membrane potentials, intracellular calcium concentrations, and NLRP3 inflammasome activation.

A New Species of the Genus Rhacophorus (Anura： Rhacophoridae) from Dabie Mountains in East China

A new species of rhacophorid of the genus Rhacophorus is described from the Dabie Mountains of west Anhui, east China. The new species, Rhacophorus zhoukaiyae sp. nov. is distinguished from its congeners by a combination of the following characters： 1） the ventral surface and front-and-rear of the femur is paler yellowish and decorated with irregular grayish blotching, and without obvious spots on the dorsum of the hand and foot webbing; 2） the outer metatarsal tubercle is small; 3） outer fingers are half-webbed and outer toes two third webbed; 4） the skin on the dorsum is smooth and without compressed warts; 5） the throat, chest and belly are pure paler yellowish; 6） the dorsal part of the fingers and toes are grayish-white; 7） the iris is golden-yellow. In addition, the phylogenetic tree showed that all the individuals of R. zhoukaiyae sp. nov. clustered into one distinct clade which suggested the validity of this species. This results could also be used to the support of species delimitation. Currently, this species is known only from mid-elevation montane evergreen forest in the Dabie Mountains of west Anhui, China.

A New Species of the Genus Protobothrops(Squamata:Viperidae:Crotalinae) from the Dabie Mountains,Anhui,China

During a biological survey in July 2011, a pit viper was collected from the Yaoluoping Nature Reserve in the Dabie Mountains, Anhui, China. The pit viper’s the total length measured 836 mm with a dorsal ground color of yellowbrown, decorated by 56 dark-brown transverse bands. The 4thsupralabial squama of the new species is separated from subocular by two squamae, and the tip of tail is orange; this combination of characters distinguish it from other species of the genus Protobothrops. Based on the body proportions, number of squamae, body color pattern, and comparing the data of its life history with those of other species of Protobothrops, we herein describe it as a new species, belonging to the genus of Protobothrops.

广东地区犬瘟热病毒血凝素基因的克隆与序列分析

【目的】对广州和东莞市宠物犬感染犬瘟热病毒(CDV)的情况进行病原鉴定和分析,为监测CDV的遗传变异情况和防治犬瘟热(CD)提供数据基础。【方法】从表现CD症状的犬只中鉴定了17份CDV阳性样本,采用RT-PCR的方法克隆得到这些野毒株的血凝素(H)基因序列,采用生物信息学方法进行序列比对分析。【结果】17株CDV的H基因核苷酸与氨基酸序列的相似性分别为97.4%~100.0%和97.5%~100.0%,与Onderstepoort、Lederle和Convac等疫苗株相比,其核苷酸与氨基酸序列相似性分别为90.3%~91.5%和89.4%~90.8%。进化树分析结果显示,17株CDV野毒株均属于AsiaⅠ型,与疫苗株的分支较远;本研究鉴定的野毒株已进化形成9个潜在的N-糖基化位点。【结论】AsiaⅠ型CDV仍为该地区的流行基因型,基因型较稳定,但与疫苗株相比形成了一定的进化距离和出现了大量的变异。因此,继续监控CDV在犬群中的进化,掌握其遗传变异状况具有重要意义。

Systematics and Species Validity of the Dabieshan Pit Viper Protobothrops dabieshanensis Huang et al. 2012: Evidence from a Mitochondrial Gene Sequence Analysis

Aimed to evaluate the phylogenetic position of the recently described Protobothrops dabieshanensis Huang et al. （2012）, phylogenic relationships of 12 species within Protobothrops based on four mtDNA gene fragments （12S RNA, 16S RNA, ND4 and Cyt b） were reconstructed in our study. The result indicates a clade composed ofP dabiesha- nensis, P. jerdonii and P xiangchengsis with strong support. The genetic distance among P dabieshanensis, P jerdonii and P xiangchengsis was much lower than other congeners. Based on the data from the phylogenetic analysis and pre- viously described morphological differences, we conclude that P dabieshanensis is a valid species with close affinities to P jerdonii and P xiangchengsis.