MCP: Multi-Chicken Pose Estimation Based on Transfer Learning.

Poultry managers can better understand the state of poultry through poultry behavior analysis. As one of the key steps in behavior analysis, the accurate estimation of poultry posture is the focus of this research. This study mainly analyzes a top-down pose estimation method of multiple chickens. Therefore, we propose the "multi-chicken pose" (MCP), a pose estimation system for multiple chickens through deep learning. Firstly, we find the position of each chicken from the image via the chicken detector; then, an estimate of the pose of each chicken is made using a pose estimation network, which is based on transfer learning. On this basis, the pixel error (PE), root mean square error (RMSE), and image quantity distribution of key points are analyzed according to the improved chicken keypoint similarity (CKS). The experimental results show that the algorithm scores in different evaluation metrics are a mean average precision (mAP) of 0.652, a mean average recall (mAR) of 0.742, a percentage of correct keypoints (PCKs) of 0.789, and an RMSE of 17.30 pixels. To the best of our knowledge, this is the first time that transfer learning has been used for the pose estimation of multiple chickens as objects. The method can provide a new path for future poultry behavior analysis.

Super-resolution fusion optimization for poultry detection: a multi-object chicken detection method.

Accurate poultry detection is crucial for studying poultry behavior using computer vision and video surveillance. However, in free-range farming environments, detecting chickens can often be challenging due to their small size and mutual occlusion. The current detection algorithms exhibit a low level of accuracy, with a high probability of false and missed detections. To address this, we proposed a multi-object chicken detection method named Super-resolution Chicken Detection, which utilizes super-resolution fusion optimization. The algorithm employs the residual-residual dense block to extract image features and used a generative adversarial network to compensate for the loss of details during deep convolution, producing high-resolution images for detection. The proposed algorithm was validated with the B1 data set and the MC1 multi-object data set, demonstrating that the reconstructed images possessed richer pixel features compared to original images, specifically it improved detection accuracy and reduced the number of missed detections. The structural similarity of the reconstructed images was 99.9%, and the peak signal-to-noise ratio was above 30. The algorithm improved the Average Precision50:95 of all You Only Look Once Version X (YOLOX) models, with the largest improvement for the B1 data set with YOLOX-Large (+6.3%) and for the MC1 data set with YOLOX-Small (+4.1%). This was the first time a super-resolution reconstruction technique was applied to multi-object poultry detection. Our method will provide a fresh approach for future poultry researchers to improve the accuracy of object detection using computer vision and video surveillance.

In free-range farming environments, accurately detecting individual chickens has been a persistent challenge for researchers. Due to mutual occlusion and limitations of camera capturing distance, existing detection algorithms have had low detection accuracy, leading to a high probability of false and missed detections. To address this problem, a multi-object chicken detection method named Super-resolution Chicken Detection (SRCD) was developed. The proposed algorithm utilized super-resolution reconstruction and You Only Look Once Version X object detection networks to achieve accurate chicken detection. Through extensive experimentation with two data sets, SRCD demonstrated its superiority in detecting chickens, reducing missed detections likely related to occlusion and the chickens' distance to the camera. Additionally, the SRCD algorithm enriched the pixel features of the chickens with minimal changes to the original images, resulting in a reconstructed image with high similarity to the original image. As a result, this method provided a practical solution for more accurate detection in small free-range farming environments, improving poultry production efficiency.

Sex Detection of Chicks Based on Audio Technology and Deep Learning Methods.

The sex detection of chicks is an important work in poultry breeding. Separating chicks of different sexes early can effectively improve production efficiency and commercial benefits. In this paper, based on the difference in calls among one-day-old chicks of different sexes, a sex detection method based on chick calls is designed. Deep learning methods were used to classify the calls of chicks and detect their sex. This experiment studies three different varieties of chicks. The short-time zero-crossing rate was used to automatically detect the endpoints of chick calls in audio. Three kinds of audio features were compared: Spectrogram, Cepstrogram and MFCC+Logfbank. The features were used as the input in neural networks, and there were five kinds of neural networks: CNN, GRU, CRNN, TwoStream and ResNet-50. After the cross-comparison experiment of different varieties of chicks, audio features and neural networks, the ResNet-50 neural network trained with the MFCC+Logfbank audio features of three yellow chick calls had the highest test accuracy of 83% when testing Three-yellow chicks' calls. The GRU neural network trained with the Spectrogram audio features of native chick calls had the highest test accuracy of 76.8% when testing Native chicks' calls. The ResNet-50 neural network trained with Spectrogram audio features of flaxen-yellow chick calls had the highest test accuracy of 66.56%when testing flaxen-yellow chick calls. Multiple calls of each chick were detected, and the majority voting method was used to detect the sex of the chicks. The ResNet-50 neural network trained with the Spectrogram of three yellow chick calls had the highest sex detection accuracy of 95% when detecting the three yellow chicks' sex. The GRU neural network trained with the Spectrogram and cepstrogram of native chick calls and the CRNN network trained with the Spectrogram of native chick calls had the highest sex detection accuracy of 90% when detecting the native chicks' sex. The Twostream neural network trained with MFCC+Logfbank of flaxen-yellow chick calls and the ResNet-50 network trained with the Spectrogram of flaxen-yellow chick calls had the highest sex detection accuracy of 80% when detecting the flaxen-yellow chicks' sex. The results of the cross-comparison experiment show that there is a large diversity between the sex differences in chick calls of different breeds. The method is more applicable to chick sex detection in three yellow chicks and less so in native chicks and flaxen-yellow chicks. Additionally, when detecting the sex of chicks of a similar breed to the training chicks, the method obtained better results, while detecting the sex of chicks of other breeds, the detection accuracy was significantly reduced. This paper provides further perspectives on the sex detection method of chicks based on their calls and help and guidance for future research.

Molecular prevalence and genetic diversity analysis of Enterocytozoon bieneusi in humans in Hainan Province, China: High diversity and unique endemic genetic characteristics.

Enterocytozoon bieneusi is a zoonotic pathogen commonly found in humans and animals all over the world. Here, we investigated the occurrence and genotype constitute of E. bieneusi among the individuals from Haikou city of Hainan, China. A total of 1,264 fecal samples of humans were collected, including 628 samples from patients with diarrhea (325 adults and 303 children) and 636 samples from the asymptomatic population (383 college students and 253 kindergarten children). E. bieneusi was detected using nested polymerase chain reaction (PCR) amplification of the internal transcribed spacer (ITS) region. A phylogenetic tree was constructed using a neighbor-joining tree construction method. The overall prevalence of E. bieneusi was 3.7% (47/1,264), while it was 5.6% in the patients with diarrhea (5.8% in adults and 5.3% in children) and 1.9% in the asymptomatic population (2.9% in college students and 0.4% in kindergarten children). The prevalence of E. bieneusi in humans with diarrhea was significantly higher than that in the asymptomatic population (χ2 = 36.9; P < 0.05). A total of 28 genotypes were identified, including ten known genotypes: CHG2 (n = 3), CHG3 (n = 5), CHG5 (n = 10), CM21 (n = 1), EbpA (n = 1), EbpC (n = 1), PigEBITS4 (n = 1), PigEBITS7 (n = 1), SHR1 (n = 4), Type IV (n = 2), and 18 novel genotypes (HNH-1 to HNH-18; one each). All these genotypes were categorized into three groups, including group 1 (n = 6), group 2 (n = 14), and group 13 (n = 8). This was the first study on the identification of E. bieneusi among humans in Hainan, China. The correlation between E. bieneusi infection and diarrhea was observed. The high diversity and distinctive distribution of E. bieneusi genotypes found in this study reflected the unique epidemic genetic characteristics of E. bieneusi in humans living in Hainan.

Study on Poultry Pose Estimation Based on Multi-Parts Detection.

Poultry pose estimation is a prerequisite for evaluating abnormal behavior and disease prediction in poultry. Accurate pose-estimation enables poultry producers to better manage their poultry. Because chickens are group-fed, how to achieve automatic poultry pose recognition has become a problematic point for accurate monitoring in large-scale farms. To this end, based on computer vision technology, this paper uses a deep neural network (DNN) technique to estimate the posture of a single broiler chicken. This method compared the pose detection results with the Single Shot MultiBox Detector (SSD) algorithm, You Only Look Once (YOLOV3) algorithm, RetinaNet algorithm, and Faster_R-CNN algorithm. Preliminary tests show that the method proposed in this paper achieves a 0.0128 standard deviation of precision and 0.9218 ± 0.0048 of confidence (95%) and a 0.0266 standard deviation of recall and 0.8996 ± 0.0099 of confidence (95%). By successfully estimating the pose of broiler chickens, it is possible to facilitate the detection of abnormal behavior of poultry. Furthermore, the method can be further improved to increase the overall success rate of verification.

Design and Implementation of Poultry Farming Information Management System Based on Cloud Database.

Aiming at breaking down the bottleneck problems of different scale of poultry farms, the low profitability of poultry farming, and backward information management in China, a safe and efficient information management system for poultry farming was designed. This system consists of (1) a management system application layer, (2) a data service layer, and (3) an information sensing layer. The information sensing layer obtains and uploads production and farming information through the wireless sensor network built in the poultry house. The use of a cloud database as an information storage carrier in the data service layer eliminates the complex status of deploying local server clusters, and it improves the flexibility and scalability of the system. The management system application layer contains many sub-function modules including poultry disease detection functions to realize the visual management of farming information and health farming; each module operates independently and cooperates with each other to form a set of information management system for poultry farming with wide functional coverage, high service efficiency, safety, and convenience. The system prototype has been tested for the performance of wireless sensor network and cloud database, and the results show that the prototype is capable of acquiring and managing poultry farming information.

LncRNA ADAMTS9-AS2 inhibits gastric cancer (GC) development and sensitizes chemoresistant GC cells to cisplatin by regulating miR-223-3p/NLRP3 axis.

The role of LncRNA ADAMTS9-AS2 in the regulation of chemoresistance of gastric cancer (GC) is largely unknown. Here we found that LncRNA ADAMTS9-AS2 was low-expressed in GC tissues and cells compared to their normal counterparts. In addition, LncRNA ADAMTS9-AS2 inhibited miR-223-3p expressions in GC cells by acting as competing endogenous RNA, and the levels of LncRNA ADAMTS9-AS2 and miR-223-3p showed negative correlations in GC tissues. Of note, overexpression of LncRNA ADAMTS9-AS2 inhibited GC cell viability and motility by sponging miR-223-3p. In addition, the levels of LncRNA ADAMTS9-AS2 were lower, and miR-223-3p was higher in cisplatin-resistant GC (CR-GC) cells than their parental cisplatin-sensitive GC (CS-GC) cells. LncRNA ADAMTS9-AS2 overexpression enhanced the cytotoxic effects of cisplatin on CR-GC cells, which were reversed by overexpressing miR-223-3p. Furthermore, LncRNA ADAMTS9-AS2 increased NLRP3 expressions by targeting miR-223-3p, and upregulation of LncRNA ADAMTS9-AS2 triggered pyroptotic cell death in cisplatin treated CR-GC cells by activating NLRP3 inflammasome through downregulating miR-223-3p. Finally, the promoting effects of LncRNA ADAMTS9-AS2 overexpression on CR-GC cell death were abrogated by pyroptosis inhibitor Necrosulfonamide (NSA). Collectively, LncRNA ADAMTS9-AS2 acted as a tumor suppressor and enhanced cisplatin sensitivity in GC cells by activating NLRP3 mediated pyroptotic cell death through sponging miR-223-3p.

Tripartite Motif 8 (TRIM8) Positively Regulates Pro-inflammatory Responses in Pseudomonas aeruginosa-Induced Keratitis Through Promoting K63-Linked Polyubiquitination of TAK1 Protein.

Pseudomonas aeruginosa (PA)-induced keratitis is a rapidly progressive ocular infectious disease that often leads to inflammatory epithelial edema, stromal infiltration, tissue destruction, corneal ulceration, and vision loss. In this study, we investigate the role of tripartite motif 8 (TRIM8) in regulating the inflammatory process of PA-induced keratitis. The expression of TRIM8 was increased in mouse corneas and in vitro-cultured macrophages after PA infection. Knockdown of the expression of TRIM8 significantly inhibited the activation of NF-κB signaling and decreased the production of pro-inflammatory cytokines both in vivo and in vitro after infected with PA. Furthermore, we investigated the potential mechanism and we found after PA infection that TRIM8 could promote K63-linked polyubiquitination of transforming growth factor ß-activated kinase 1 (TAK1), leading to the activation of TAK1 and enhanced inflammatory responses. Taken together, we demonstrated that TRIM8 has pro-inflammatory effect on PA-induced keratitis and suggest TRIM8 as a potential therapeutic target for keratitis.

Pathogenicity, Transmission and Antigenic Variation of H5N1 Highly Pathogenic Avian Influenza Viruses.

H5N1 highly pathogenic avian influenza (HPAI) was one of the most important avian diseases in poultry production of China, especially in Guangdong province. In recent years, new H5N1 highly pathogenic avian influenza viruses (HPAIV) still emerged constantly, although all poultry in China were immunized with H5N1 vaccinations compulsorily. To better understand the pathogenicity and transmission of dominant clades of the H5N1 HPAIVs in chicken from Guangdong in 2012, we chose a clade 7.2 avian influenza virus named A/Chicken/China/G2/2012(H5N1) (G2) and a clade 2.3.2.1 avian influenza virus named A/Duck/China/G3/2012(H5N1) (G3) in our study. Our results showed that the chickens inoculated with 10(3) EID50 of G2 or G3 viruses all died, and the titers of virus replication detected in several visceral organs were high but different. In the naive contact groups, virus shedding was not detected in G2 group and all chickens survived, but virus shedding was detected in G3 group and all chickens died. These results showed that the two clades of H5N1 HPAIVs had high pathogenicity in chickens and the contact transmission of them was different in chickens. The results of cross reactive HI assay showed that antigens of G2 and G3 were very different from those of current commercial vaccines isolates (Re-4, Re-6, and D7). And to evaluate the protective efficacy of three vaccines against most isolates form Guangdong belonging to clade 2.3.2.1 in 2012, G3 was chosen to challenge the three vaccines such as Re-4, Re-6, and D7. First, chickens were immunized with 0.3 ml Re-4, Re-6, and D7 inactivated vaccines by intramuscular injection, respectively, and then challenged with 10(6) EID50 of G3 on day 28 post-vaccination. The D7 vaccine had 100% protection against G3 for chickens, the Re-6 vaccine had 88.9%, and the Re-4 vaccine only had 66.7%. Our results suggested that the D7 vaccine could prevent and control H5N1 virus outbreaks more effectively in Guangdong. From the above, it was necessary to conduct continuously epidemiological survey and study the pathogenicity and antigenic variation of avian influenza in Southern China.

Magnetic nanosphere-guided site-specific delivery of vascular endothelial growth factor gene attenuates restenosis in rabbit balloon-injured artery.

OBJECTIVE: New and efficient strategies to protect endothelium or to enhance endothelial regrowth are important for treatment of restenosis after percutaneous transluminal angioplasty. Magnetic DNA microspheres are used to accelerate vascular endothelial growth factor (VEGF) re-endothelialization and to attenuate intimal hyperplasia in balloon-injured artery. This study aimed to assess DNA-gelatin magnetic nanospheres containing VEGF expression plasmids in vascular restenosis attenuation. METHODS: Ninety-six rabbits underwent balloon injury and were randomly divided for gene transfer with naked VEGF plasmids (NAK group), magnetic VEGF microspheres (MIC group), and LacZ (CON group: naked LacZ plasmid and LacZ nanosphere subgroups). Serum and tissue VEGF levels were measured. Also, the ratios of intima area to media area were determined to assess neointima formation. RESULTS: Microsphere gene delivery through the artery by a magnet resulted in VEGF overexpression in transfected arterial segments. Tissue VEGF integral optical densities were significantly increased in MIC rabbits compared with NAK animals. Serum VEGF was below detection in all animals. X-Gal staining showed higher transfection efficiency in the CON group. The impact of neointimal thickening was evaluated by light microscopy as the ratio of intima area to media area in cross sections. Significant differences in the ratio of intima area to media area were obtained between the NAK group (0.12 ± 0.02, 0.41 ± 0.03, 0.61 ± 0.05, and 0.72 ± 0.04 at 1, 2, 3, and 4 weeks, respectively) and the MIC group (0.06 ± 0.03, 0.20 ± 0.05, 0.25 ± 0.04, and 0.26 ± 0.03 at 1, 2, 3, and 4 weeks, respectively) at 2, 3, and 4 weeks (P < .05). CONCLUSIONS: Intra-arterial VEGF gene delivery by magnetic microspheres significantly increased DNA stability, transfection efficiency, and targeting specificity, resulting in exogenous VEGF overexpression and attenuated intimal hyperplasia in balloon-injured artery.

The first report of human-derived G10 genotype of Echinococcus canadensis in China and possible sources and routes of transmission.

Cystic echinococcosis (CE) is one of the most important parasitic zoonoses. 10 distinct genotypes, designated G1-G10 genotypes of Echinococcus granulosus sensu lato (s.l.), have been split into 4 species: Echinococcus granulosus sensu stricto (s.s.) (G1-G3), Echinococcus equinus (G4), Echinococcus ortleppi (G5) and Echinococcus canadensis (G6-G10); Echinococcus felidis has also been suggested as a sister taxon of E. granulosus s.s. recently. Four genotypes belonging to two species (G1 and G3 genotypes of E. granulosus s.s., and G6 and G7 genotypes of E. canadensis) have been identified in humans and animals in China. In the present study, a human-derived hydatid cyst from a patient in northeastern China's Heilongjiang Province was identified as G10 genotype of E. canadensis based on mitochondrial cytochrome c oxidase subunit I (cox1), cytochrome b (cytb) and NADH dehydrogenase subunit 1 (nad1) genes. Homology analysis showed the cox1 gene sequence of G10 genotype of E. canadensis had 100% homology with those from wolves in Mongolia and from a moose in Russia. The cytb and nad1 gene sequences of G10 genotype of E. canadensis had 100% homology with the complete sequence from a moose in Finland at an amino acid level. The infection source of the CE patient here might be primarily attributable to wolves. This is the first report of G10 genotype of E. canadensis in a human in China. The finding of G10 genotype of E. canadensis in China shows that this genotype possibly has a more wide geographical distribution than previously considered.

Human cystic echinococcosis in Heilongjiang Province, China: a retrospective study.

BACKGROUND: Cystic echinococcosis (CE) is one of emerging zoonotic parasitic diseases throughout the world, having significant medical and economic importance in developing countries. The western and northwestern China is considered as CE endemic areas. In northeastern China's Heilongjiang Province, the increasing number of sporadic human CE cases has attracted more and more attention. The aims of the present study were to understand the clinical characteristics of human CE in the investigated area and to compare the coincidence rates of CT, ultrasound and serological test against the histopathology results among CE patients. METHODS: Hospital data of 183 human CE cases in the period from January 2004 to July 2013 were collected from the two largest hospitals in Heilongjiang Province. Clinical data were analyzed, including age, gender, occupation and living residence of CE patients and localization, size and number of CE cysts as well as the diagnosis methods of CE before operation. RESULTS: The results revealed that the incidence of CE reached a peak in the age group of 41-50 years. Among the 183 CE patients, the females were observed to have a higher percentage of CE patients (60.66%, 111/183) than males (39.34%, 72/183). The majority of CE patients were farmers, followed by workers, employees, public servants, students and so on. CE cysts were most commonly found in the livers, with a 30 cm cyst in diameter being detected. CT showed the highest coincidence rate (96.64%) for hepatic CE among the three common diagnosis methods (CT, ultrasound imagine and serological test) compared against the histopathology results. CONCLUSIONS: This is the first retrospective analysis of human CE cases in Heilongjiang Province in recent ten years. Clinical characteristics of human CE were described here. CT appeared to be the most effective diagnosis method for hepatic CE.

Genetic characterization of human-derived hydatid cysts of Echinococcus granulosus sensu lato in Heilongjiang Province and the first report of G7 genotype of E. canadensis in humans in China.

Cystic echinococcosis (CE) caused by the larval stage of Echinococcus granulosus sensu lato (s.l.) is one of the most important zoonotic parasitic diseases worldwide and 10 genotypes (G1-G10) have been reported. In China, almost all the epidemiological and genotyping studies of E. granulosus s.l. are from the west and northwest pasturing areas. However, in Heilongjiang Province of northeastern China, no molecular information is available on E. granulosus s.l. To understand and to speculate on possible transmission patterns of E. granulosus s.l., we molecularly identified and genotyped 10 hydatid cysts from hepatic CE patients in Heilongjiang Province based on mitochondrial cytochrome c oxidase subunit I (cox1), cytochrome b (cytb) and NADH dehydrogenase subunit 1 (nad1) genes. Two genotypes were identified, G1 genotype (n = 6) and G7 genotype (n = 4). All the six G1 genotype isolates were identical to each other at the cox1 locus; three and two different sequences were obtained at the cytb and nad1 loci, respectively, with two cytb gene sequences not being described previously. G7 genotype isolates were identical to each other at the cox1, cytb and nad1 loci; however, the cytb gene sequence was not described previously. This is the first report of G7 genotype in humans in China. Three new cytb gene sequences from G1 and G7 genotypes might reflect endemic genetic characterizations. Pigs might be the main intermediate hosts of G7 genotype in our investigated area by homology analysis. The results will aid in making more effective control strategies for the prevention of transmission of E. granulosus s.l.

Calcium sensing receptor promotes cardiac fibroblast proliferation and extracellular matrix secretion.

AIMS: Calcium-sensing receptor (CaR) acts as a G protein coupled receptor that mediates the increase of the intracellular Ca(2+) concentration. The expression of CaR has been confirmed in various cell types, including cardiomyocytes, smooth muscle cells, neurons and vascular endothelial cells. However, whether CaR is expressed and functions in cardiac fibroblasts has remained unknown. The present study investigated whether CaR played a role in cardiac fibroblast proliferation and extracellular matrix (ECM) secretion, both in cultured rat neonatal cardiac fibroblasts and in a model of cardiac hypertrophy induced by isoproterenol (ISO). METHODS AND RESULTS: Immunofluorescence, immunohistochemistry and Western blot analysis revealed the presence of CaR in cardiac fibroblasts. Calcium and calindol, a specific activator of CaR, elevated the intracellular calcium concentration in cardiac fibroblasts. Pretreatment of cardiac fibroblasts with calhex231, a specific inhibitor of CaR, U73122 and 2-APB attenuated the calindol- and extracellular calcium-induced increase in intracellular calcium ([Ca(2+)]i). Cardiac fibroblast proliferation and migration were assessed by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), cell count and the cell scratch assay. ECM production was detected by expression of matrix metalloproteinase-3 and -9 (MMP-3 and -9). Activation of CaR promoted cardiac fibroblast proliferation and migration and ECM secretion. More importantly, calhex231, suppressed cardiac fibroblast proliferation and migration and MMP-3 and -9 expression. To further investigate the effect of CaR on cardiac fibrosis, a model of ISO-induced cardiac hypertrophy was established. Pretreatment with calhex231 prevented cardiac fibrosis and decreased the expression of MMP-3 and -9 expression. CONCLUSIONS: Our results are the first report that CaR plays an important role in Ca(2+) signaling involved in cardiac fibrosis through the phospholipase C- inositol 3,4,5 phosphate (PLC-IP3) pathway.

Protective effects of Purendan superfine powder on retinal neuron apoptosis in a rat model of type 2 diabetes mellitus.

This study sought to investigate the effects of Purendan superfine powder comprised of Momordica charantia, Radix Ginseng, and Radix Salviae Miltiorrhiae on neuronal apoptosis and expression of bcl-2, bax, and caspase-3, which are retinal apoptosis-associated factors in rats with diabetes mellitus induced by continuous intraperitoneal injection of streptozotocin. The results showed that Purendan superfine powder could upregulate the expression of bcl-2 protein and mRNA, and downregulate the expression of bax and caspase-3 in the retina of diabetes mellitus rats. In addition, Purendan superfine powder was shown to reduce the number of apoptotic neurons. Our experimental findings indicate that Purendan superfine powder can inhibit neuronal apoptosis in the retina of diabetes mellitus rats and has protective effects on diabetic retinopathy.

Vascular endothelial growth factor gene delivery by magnetic DNA nanospheres ameliorates limb ischemia in rabbits.

BACKGROUND: Critical limb ischemia often leads to disability and limb loss. Vascular endothelial growth factor (VEGF), delivered either as recombinant protein or as gene therapy, has been shown to promote arteriogenesis and angiogenesis in animal models of limb ischemia. However, most of the studies used a nonspecific targeting system. MATERIALS AND METHODS: Magnetic DNA nanospheres containing expression plasmids encoding VEGF were synthesized, and their morphology, magnetropism, and stability were analyzed. The magnetic DNA nanospheres were administrated via an artery into a rabbit limb ischemia model. The expression of VEGF and vascularization were examined by immunohistochemistry. The angiography was taken to evaluate arteriogenesis. RESULTS: Magnetic DNA nanospheres were very stable and showed a high magnetropism. Gene delivery of such nanospheres via artery under a magnetic field led to the overexpression of VEGF in situ. The capillary density and capillary to muscle fiber ratio were doubled compared with those of the control animals. The arteriogenesis also was promoted in VEGF gene therapy group compared with controls but at later interval than capillary angiogenesis. CONCLUSIONS: Our results suggest that intra-arterial VEGF gene delivery by magnetic DNA nanosphere promotes angiogenesis and arteriogenesis and presents a potent therapeutic strategy for critical limb ischemia.

[Study of mechanism on immune response in goats by different immunization with Helicobacter pylori and analyse of its prospects for using].

OBJECTIVES: To observe the effect of three different routes on the induction of immunogenicity to whole bacterial antigen of Vac A+ Helicobacter pylori strain (NCTC11637) and explore its mechanism. METHODS: Two milk goats and one pregnant goat were immunized by different routes respectively with whole bacterial of H. pylori (6 x 10(9) cfu/ml) cultivated by solid culture medium. At the 1st, 14th, 21st, 28th day, two goats were immunized four times by intranasal or subcutaneous injection. The other pregnant goat was immunized four times at a interval of two weeks before and after one month of lamb birth by muscular injection. Serum and milk samples were collected and assayed by indirect enayme-linked immunosorbent assay (ELISA). The levels of anti-Hp of IgG and IgA in serum and milk were determined by reading the optical density (A). RESULTS: Three immune routes all induced systemic immune response. The optical density (A) of ELISA proved that the specific IgG in serum increased while IgA didn't increase significantly, and that the specific IgA and IgG in milk all increased to a greater or lesser extent compared with control group. CONCLUSION: Three immune routes all induce systemic immune response, resulting in increases of anti-Hp of IgG/IgA in milk and IgG in serum. Among them, intranasal inoculation can induce systemic immune response and local immune response in different mucosal sites, which may be a safe and effective immunization route.