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ObjectiveHigh-precision localization technology serves as the crucial foundation in enabling the autonomous navigation operations of intelligent agricultural robots. However, the traditional global navigation satellite system (GNSS) localization method faces numerous limitations, such as tree shadow, electromagnetic interference, and other factors in the agricultural environment brings challenges to the accuracy and reliability of localization technology. To address the deficiencies and achieve precise localization of agricultural robots independent of GNSS, a localization method was proposed based on the fusion of three-dimensional light detection and ranging (LiDAR) data and inertial measurement unit (IMU) information to enhance localization accuracy and reliability.MethodsLiDAR was used to obtain point cloud data in the agricultural environment and realize self-localization via point cloud matching. By integrating real-time motion parameter measurements from the IMU with LiDAR data, a high-precision localization solution for agricultural robots was achieved through a specific fusion algorithm. Firstly, the LiDAR-obtained point cloud data was preprocessed and the depth map was used to save the data. This approach could reduce the dimensionality of the original LiDAR point cloud, and eliminate the disorder of the original LiDAR point cloud arrangement, facilitating traversal and clustering through graph search. Given the presence of numerous distinct crops like trees in the agricultural environment, an angle-based clustering method was adopted. Specific angle-based clustering criteria were set to group the point cloud data, leading to the segmentation of different clusters of points, and obvious crops in the agricultural environment was effectively perceived. Furthermore, to improve the accuracy and stability of positioning, an improved three-dimensional normal distribution transform (3D-NDT) localization algorithm was proposed. This algorithm operated by matching the LiDAR-scanned point cloud data in real time with the pre-existing down sampled point cloud map to achieve real-time localization. Considering that direct down sampling of LiDAR point clouds in the agricultural environment could result in the loss of crucial environmental data, a point cloud clustering operation was used in place of down sampling operation, thereby improving matching accuracy and positioning precision. Secondly, to address potential constraints and shortcomings of using a single sensor for robot localization, a multi-sensor information fusion strategy was deployed to improve the localization accuracy. Specifically, the extended Kalman filter algorithm (EKF) was chosen to fuse the localization data from LiDAR point cloud and the IMU odometer information. The IMU provided essential motion parameters such as acceleration and angular velocity of the agricultural robot, and by combining with the LiDAR-derived localization information, the localization of the agricultural robot could be more accurately estimated. This fusion approach maximized the advantages of different sensors, compensated for their individual limitations, and improved the overall localization accuracy of the agricultural robot.Results and DiscussionsA series of experimental results in the Gazebo simulation environment of the robot operating system (ROS) and real operation scenarios showed that the fusion localization method proposed had significant advantages. In the simulation environment, the average localization errors of the proposed multi-sensor data fusion localization method were 1.7 and 1.8 cm, respectively, while in the experimental scenario, these errors were 3.3 and 3.3 cm, respectively, which were significantly better than the traditional 3D-NDT localization algorithm. These findings showed that the localization method proposed in this study could achieve high-precision localization in the complex agricultural environment, and provide reliable localization assistance for the autonomous functioning of agricultural robots.ConclusionsThe proposed localization method based on the fusion of LiDAR data and IMU information provided a novel localization solution for the autonomous operation of agricultural robots in areas with limited GNSS reception. Through the comprehensive utilization of multi-sensor information and adopting advanced data processing and fusion algorithms, the localization accuracy of agricultural robots could be significantly improved, which could provide a new reference for the intelligence and automation of agricultural production.

Objective To compare the diagnostic values of transperineal and transrectal ultrasound-guided prostate puncture for prostate cancer in patients with gray / sub-gray zone of prostate-specific antigen （PSA）. Methods A total of 110 samples were collected from patients with PSA≤20 ng/mL who underwent transperineal and transrectal ultrasound-guided prostate puncture. All patients were divided into two groups according to different patterns of prostate puncture. The detection rates of prostate cancer and CsPCa of patients with PSA in the gray/ sub-gray zone were compared between two different puncture methods. Surgical conditions and degree of pain were compared between two groups，including operation time，bleeding，intra-operative and post-operative VAS pain scores. The incidence of complications was also compared between two groups，including pain，fever and hematuria，etc. The probability of complications was assessed in two groups. Results The detection rate of prostate cancer in the transperineal ultrasound-guided prostate puncture was higher than that in the transrectal prostate puncture （P<0.05）. Further subgroup analyses showed no statistical significance in the detection rates of prostate cancer in patients with PSA gray zone between two groups （P>0.05），but patients with PSA sub-gray zone in the transperineal ultrasound-guided prostate puncture group had a higher detection rate （P<0.05）. PSA sub-gray zone CsPCa in the PSA sub-gray zone accounted for a higher proportion of patients with positive puncture than that in the PSA gray zone （P<0.05）. There was no significant difference between two different puncture methods in terms of puncture time，bleeding，degree of pain and incidence of complications （all P>0.05）. Conclusions For patients with PSA gray zone，there is no significant difference in the detection rate of prostate cancer between two modalities. Nevertheless，for patients with PSA between 10-20 ng/mL sub-gray zone，transperineal ultrasound-guided prostate puncture yields better diagnostic and application value compared with transrectal ultrasound-guided prostate puncture.

【Objective】 Predicting change in groundwater table in response to natural and anthropogenic influence is a prerequisite in groundwater resource management. This paper compares several methods commonly used in studying groundwater dynamics based on the shallow groundwater exploration in plain areas in central China. 【Method】 The study was based on shallow groundwater resources in Puyang City in Henan province. We analyzed the groundwater dynamics in the region using three methods: water balance method, dewatering-volume method, and time series method. 【Result】 The time series method is applicable only to areas where groundwater development and utilization do not show considerable change with time. In contrast, the water balance method and dewatering volume method are more mechanistic, providing more accurate results as they consider potential changes in groundwater usage and management. As the dewatering volume method divides the studied region into subunits based on administrative regions rather than hydrogeological conditions, it could lead to significant differences when compared to measurements from monitoring boreholes. The water balance method considers factors such as current groundwater development, hydrogeological conditions and groundwater management measures, and it is hence more accurate for analyzing groundwater dynamics. 【Conclusion】 Results calculated by the improved groundwater balance method proposed in this paper are consistent with ground-truth data. It is more reliable for evaluating the response of shallow groundwater table to change in management and usages in plain areas.

ObjectiveBy analyzing the food consumption structure and carbon emissions among Chinese residents from 2014 to 2020， this study aims to provide scientific support for guiding residents towards adopting sustainable consumption patterns and achieving the dual carbon goals of carbon peak and carbon neutrality.MethodsThe study focuses on food consumption and extends to food-related carbon emissions. By using comprehensive food carbon emission coefficients and the life cycle assessment method， both direct and indirect carbon emissions caused by food consumption among Chinese residents were calculated， and the evolving trend of food-related carbon emissions is explored.ResultsThe food structure of Chinese residents increases year by year with the animal food consumption， and the greenhouse gas emissions also increase significantly. Under the hypothetical scenarios assuming gradual adjustments of food consumption structure to meet dietary guidelines by 2030， 2040 and 2050， the total greenhouse gas emissions are projected to peak in 2030， 2040 and 2049， respectively， with certain increases compared to the current level.ConclusionThe adjustment of food consumption structure in China has brought great pressure on carbon emission reduction. Based on the above results and conclusions， reasonable policy implications are proposed.

BACKGROUND The North Henan Plain is located in the middle and lower reaches of the Yellow River, and both high arsenic and fluoride groundwater exist. However, the coexistence mechanism of arsenic and fluoride in this sporadic distribution area is unclear. OBJECTIVES To investigate the spatial distribution characteristics and formation mechanism of arsenic and fluoride in shallow groundwater in the North Henan Plain. METHODS 332 groups of shallow groundwater samples were collected and analyzed in the North Henan Plain. Atomic fluorescence spectrometry was used to determine arsenic content, and ion chromatography and inductively coupled plasma emission spectroscopy were used to determine the content of fluoride and other cation-anions. Based on the spatial distribution of arsenic and fluoride, combined with hydrochemical diagrams and factor analysis, three main factors affecting the evolution of groundwater in this area were extracted, and the formation mechanism of high arsenic and high fluoride groundwater in this area was discussed. RESULTS The concentrations of arsenic and fluoride in groundwater were 0.0001-0.1900mg/L and 0.13-4.94mg/L, respectively. The high-arsenic groundwater was mainly distributed in the vertical depth of 15-80m in the front alluvial-diluvial depression of Taihang Mountain and the Yellow River flood fan. The high-fluoride groundwater was mainly distributed in the vertical depth of 7-100m in the modern channel influence zone of the Yellow River. Evaporation and concentration, mineral dissolution/desorption, and redox environment were the main factors controlling the evolution of groundwater in this area. Fluoride had loads of 0.214 and 0.743 in factor F1 (evaporation and concentration) and F2 (mineral dissolution/desorption), respectively. High concentration of F appeared in groundwater with low concentration of Ca2+, and the concentration of F was positively correlated with ρ(Na+)/[ρ(Na+)+ρ(Ca2+)]. The strong evaporation and concentration in the modern channel influence zone of the Yellow River contributed to the dissolution of fluorine-containing minerals, and the irrigation of the Yellow River water increased the concentration of Na+ in groundwater, which further enhanced the dissolution. Arsenic had a load of 0.728 in factor F3 (redox environment). Arsenic was positively correlated with Fe2+ and NH4+, and negatively correlated with NO3- and SO42-. The lower Eh corresponds to the higher arsenic concentration. The reductive environment in the front alluvial-diluvial depression of Taihang Mountain and the Yellow River flood fan was favorable for the reductive dissolution of arsenic containing iron oxides/hydroxides, resulting in the formation of the high-arsenic groundwater. The desorption of arsenate/arsenite/fluoride in the form of anions on the mineral surface caused by the increase of pH value was favorable for the coexistence of arsenic and fluoride in groundwater. However, the correlation between arsenic and fluoride in groundwater in this area was not significant. The high concentration of calcium ions in the high arsenic region was not conducive to the enrichment of fluoride. In contrast, the weak reducing conditions in the high fluoride region were not conducive to the dissolution of arsenic-containing iron oxides/hydroxides. CONCLUSIONS The results clarify the coexistence mechanism of arsenic and fluoride in the North Henan Plain, and enrich the theoretical system of co-contamination of groundwater with high arsenic and fluoride.

ObjectiveA “multi-integration， effective control， and intelligent guarantee” solution was built for the application scenarios of food safety emergency support for the Winter Olympics.MethodsWith emergency risk assessment method， the intelligent quick screening device and method for harmful factors in the food supply chain were integrated based on the knowledge map， and big data， artificial intelligence and mobile internet technologies were used to construct an intelligent management and control system for risk early warning of the food supply chain in the Olympics to realize that the emergency plan for different links and different harmful factors in the food supply chain was intelligently generated，which were precisely pushed to users at different levels of the system.ResultsUsing intelligent rapid screening technology and prediction and early warning technology， it has been practical application in the food safety emergency drills for food safety emergencies in the Beijing Winter Olympics and Winter Paralympics （Zhangjiakou Area） 2022， and guarantee of food safety for Olympic food companies in the Winter Olympics test competitions， and the official competitions.ConclusionTaking food safety risk analysis and data governance as the main line， intelligent integration of quick screening technology as a means， and intelligent risk prediction and intelligent early warning interaction as a link， the systematic perception， targeted identification， and accurate verification of harmful factors are promoted in the food supply chain to improve the food safety emergency support capability for major events.

ObjectiveTo study the research and application of foodborne disease risk prevention.MethodsTaking the knowledge graph as the carrier， through knowledge extraction and relationship mining on the technical guide for epidemiological investigation of food safety accidents and internet data， the association between the pathogenic factors of common foodborne diseases and clinical manifestations， incubation period， susceptible population and food sources was deeply explored and obtained.ResultsThe knowledge graph of foodborne diseases was constructed， including 390 nodes and 1 375 edges. On this basis， a knowledge question answering system was constructed to realize the automatic acquisition of answers to the clinical manifestations， susceptible populations， possible food sources and biological sample collection requirements of designated foodborne disease pathogenic factors.ConclusionThis research innovatively proposed the design of foodborne disease risk prevention system based on knowledge graph technology. Meanwhile， the constructed foodborne disease knowledge graph can effectively solve unclear definition of key information and serious data islands in the related detection of pathogenic factors. The question answering system built on this basis was of great practical importance to improve the public’s knowledge of foodborne disease risks and to correct the food safety knowledge and operational behaviors of health care professionals related to foodborne diseases.

Objective To investigate the effects of eukaryotic elongation factor-2 kinase (eEF2K) gene transfection and sodium tanshinone Ⅱ A sulfonate (STS) on proliferation, invasion and migration of lung adenocarcinoma cell line A549 and its mechanism. Methods The optimal concentration of STS was screened out by preliminary experiment. A549 cells were divided into siRNA-NC group (transfected with siRNA-NC), siRNA-eEF2K group (transfected with siRNA-eEF2K), siRNA-NC+STS group (treated with 10 μg/mL STS after siRNA-NC) and siRNA-eEF2K+STS group (treated with 10 μg/mL STS after siRNA-eEF2K).The survival rate, clone formation, cell counting of transmembrane cells, migration and apoptosis of A549 cells were detected by CCK-8 method, clone formation test, Transwell chamber test, scratch test and flow cytometry. The protein expression levels of protein kinase B (AKT), phosphorylation (p)-AKT, Ki67, matrix metalloproteinase-2 (MMP-2) and B-cell lymphoma-2 genes (Bcl-2) were detected by Western blot. Results Compared with those in siRNA-NC group, the cell survival rate, clone formation rate, number of transmembrane cells, migration rate and p-AKT, Ki67, MMP-2 and Bcl-2 in proteins siRNA-eEF2K+STS group were significantly lower, while the apoptosis rate was significantly higher (P＜0.05). The siRNA-eEF2K+STS group had the largest range of changes in the above indexes. Conclusions Silencing eEF2K combined with STS can synergistically inhibit the proliferation, invasion and migration of A549 cells and promote cell apoptosis. The mechanism is potentially related to the inhibition of protein expression of p-AKT, Ki67, MMP-2 and Bcl-2.

BACKGROUND Arsenic content in shallow groundwater of the Yellow River alluvial fan plain exceeds the standard. A comprehensive understanding of the arsenic enrichment mode driven by the nitrogen cycle of shallow groundwater in the northern Henan Plain is essential for the sustainable use of groundwater resources and the health of residents. The main area of the North Henan plain is the Yellow River alluvial fan plain. The sedimentary environment in the northern Henan plain is complex, and is influenced by the Yellow River breach, diversion and oscillation, as well as alluvial diluvial in the mountainous area around the basin. The distribution, migration and release mechanism of As, NH4+ and NO3- are quite different under different sedimentary environment conditions. The joint enrichment mechanism of the three is still unclear, which is worth further study. OBJECTIVES To investigate the effect of nitrogen cycling on arsenic migration and enrichment in groundwater in the Northern Henan Plain. METHODS 513 shallow groundwater samples were collected from the northern Henan Plain. Atomic fluorescence spectroscopy was used to determine the arsenic content, atomic absorption spectroscopy and ion chromatography, and other methods for major and trace element analysis. The correlation between nitrate, ammonia nitrogen and arsenic was investigated, and the influence of nitrogen cycle on the migration and enrichment of arsenic in groundwater was studied. RESULTS The over-standard rate of arsenic concentration in shallow groundwater in the study area was 17.3%. The occurrence and transformation modes of nitrogen under different depositional environmental conditions were important driving factors for arsenic enrichment. A large amount of NO3- was produced by nitrification in the groundwater of the alluvial fan in the piedmont alluvial fan, which had the average concentration of 9.3mg/L and was the highest in the district. At the same time, the concentration of arsenic was the lowest in the district, with the average of 1.3μg/L. The good negative correlation between NO3- and As concentration indicated that nitrification produced a large amount of NO3-, which was not conducive to the dissolution of arsenic-containing iron oxide. The depressions in front of the alluvial fan and the Yellow River crater fan with higher NH4+ content were the gathering places of high-arsenic groundwater. The average concentration of arsenic in groundwater from these two areas was 49.7μg/L and 18.9μg/L, respectively, and the exceeding rate reached 87.5% and 71.4%. The good positive correlation between arsenic content and NH4+ in groundwater indicated that the process of denitrification and dissimilative reduction of nitric acid to ammonium (DNRA) consumed NO3- in groundwater and generated a large amount of NH4+, which promoted the reduction and dissolution of iron oxides that adsorbed arsenic, forming an environment rich in arsenic. CONCLUSIONS The nitrogen cycle plays an important role in the migration and enrichment of arsenic. The migration and enrichment mode of arsenic driven by the nitrogen cycle provides a scientific basis for the treatment and supervision of groundwater with high arsenic content.

BACKGROUND A typical irrigation area of the downstream Yellow River (Henan) is an important agricultural planting area in the northern Henan Plain. The shallow water quality in this area is generally poor, which is often used for crop irrigation or livestock drinking, posing a risk to human health. Therefore, the study on the variation characteristics and mechanism of arsenic and fluorine content in groundwater in this area will help to improve the level of understanding of pollution in this area. OBJECTIVES To investigate the spatial variation characteristics and mechanism of arsenic and fluorine concentration. METHODS Based on 327 groups of shallow groundwater samples collected in 2010 and 2020 in the irrigated area, the distribution of arsenic and fluorine in the groundwater of the irrigated area was analyzed, and compared to the evolution characteristics of arsenic and fluorine in the shallow groundwater of the irrigated area in the last ten years. RESULTS In 2020, the number of samples with high arsenic concentration (>10μg/L) and high fluorine concentration (>1mg/L) in shallow groundwater accounted for 26.1% and 26.06%, respectively. The high-arsenic groundwater was distributed in the sedimentary environments with interbedding structure of sediment, such as the Taihang Mountain depression and the Yellow River alluvial plain, with strong reducibility, poor groundwater runoff and strong cation exchange, resulting in high concentration of Ca2+ in the environment. Arsenic concentration increased in 31.8% of water samples and decreased in 36.7% of water samples in the last ten years. The increase of arsenic content was caused by the dissolution and release of manganese oxide due to the enhanced reducibility of groundwater. The variation of water level caused by agricultural irrigation and water replacement in different areas during the last ten years was a potential factor of arsenic concentration change. The high-fluorine groundwater was mainly distributed along the Yellow River in Xinxiang and Puyang, Henan Province. The concentration of fluorine ion was affected by the dissolution of calcareous minerals such as fluorite in the sediments, which made the high-fluorine groundwater appear in the low-calcium environment. In the last ten years, the study area with decreased fluorine ion concentration accounted for 60.2%, whereas areas with increased fluorine ion concentration accounted for 32.1%. The overall change trend was good, but the fluorine ion concentration in the high fluorine area continued to increase. Changes in fluorine concentration were also affected by changes in Ca2+, with further increases (decreases) as Ca2+ concentration decreased (increased). Elevated fluorine in groundwater was distributed along the Yellow River, affected greatly by the Yellow River water supply. So, the groundwater flow condition was good and the cation exchange was weakened, reducing the Ca2+ content. At this time, the arsenic content in the groundwater was less affected by the environment, so the study area with fluorine increase showed an inverse relationship between arsenic and fluorine in terms of regional distribution and evolution. CONCLUSIONS This study will provide support for rational utilization of groundwater in the study area.

Aiming at the complicated adjustment, high maintenance cost and safety problems of elevator door motor, an adjustment-free elevator door motor control system was designed, which consists of a hardware platform using a Cortex-M3 processor LPC1752 with a motor-controlled pulse width modulation (PWM) module and a quadrature encoder interface (QEI), a Mitsubishi intelligent power module (IPM) module, a high accuracy rotary encoder and a general three-phase asynchronous electric motor with frequency modulation and a software control system based on an expert database built out of a variety of experiments. An accurate comparison was automatically made between the parameter values of position, speed and current and the expert database to help the incremental PID tuning, thus selflearning of door width and self-adjustment of running status can be realized. The results indicate that the system can be powered to bring the elevator door into quick operation and smooth running without presetting corresponding parameter values. [ABSTRACT FROM AUTHOR]

According to the various formats cause the phenomenon of audio file's watermarking embedding becomes complex and complicated, this paper used FFMPEG to make the various formats transform into a unity format to embed the watermarking which improved the efficiency and result, it selected improved quantizing discrete cosine transform algorithm to embed the watermarking. First, it converted the binary image into the one dimensional sequence and modulated the sequence with scrambling encryption and polarization operation, then performed the digital audio signal the quantizing DCT transform, according to each different subparagraphs energy grade, it used different embedding strength and selected four different levels coefficients, repeated embedding. In the experimental simulation results, there doesn' t exist obvious differences between the transformed audio signal and the original audio signal, the watermark algorithm is imperceptible to digital audio signal such as Gauss white noise interfere, low pass filter and loss compression. [ABSTRACT FROM AUTHOR]

To predict failure pattern of masonry wall panel more precisely, this paper improved the discrete series cellular automata (CA) model of masonry wall panels, and put forward two new methods for calculating state values in the CA model. One was the physical CA model based on the finite element analysis of masonry wall panels, the other is the analytical CA model based on the propagating appearance of boundary effect to zones within masonry wall panels. Both new models were verified by examples. The concept of relative variation coefficient was proposed and applied to the discrete series CA model and the analytical CA model. By discussion of the application range of the relative variation parameter, a fundamental technique was presented to predict a serious possible failure pattern of a new masonry wall panel on the base of only one experimental masonry wall panel. Experimental results verified the validity of the method proposed in this paper. [ABSTRACT FROM AUTHOR]

Objective: To study the effect and safety of microscopic varicocele cluster ligation (MVCL)., Methods: We selected 28 patients undergoing bilateral microscopic spermatic vein ligation in Xuzhou Central Hospital from July 2021 to June 2022. Using the computerized randomization method, we performed MVCL or microscopic varicocele ligation (MVL) for the right or the left spermatic cord, respectively. We recorded the operation time, intraoperative blood loss, the numbers of the spermatic veins ligated and the arteries and lymphatic vessels preserved in each surgical side. A surgeon unaware of the surgical approach on the operative side collected the Visual Analogue Scale (VAS) pain scores, nodular foreign body sensation, relief of scrotal cramps, complications, and long-term recurrence from the patients., Results: Compared with the MVL group, the MVCL group showed significantly shorter time for spermatic vein ligation (［56.21±13.96］ vs ［31.43±10.13］ min, P<0.01), lower VAS scores on the 1st postoperative day (P <0.05) and a lower incidence of intra-scrotal nodular foreign body sensation in the 1st postoperative month (P <0.05). There were no statistically significant differences in the intraoperative blood loss, numbers of spermatic veins ligated and arteries and lymphatic vessels preserved, VAS scores at 3 and 7 postoperative days, incidence of complications and long-term recurrence between the two groups (P >0.05)., Conclusion: MVCL is superior to MVL in reducing the time of spermatic vein ligation and improving the efficiency, efficacy and safety of the procedure, and therefore worthy of clinical promotion.

Objective: To investigate the effect of docosahexaenoic acid (DHA) on invasiveness of aflatoxin B1 (AFB1)-induced hepatocellular carcinoma cells in vitro., Methods: HepG2.2.15 cells were exposed to different concentrations of AFB1 and DHA plus AFB1. The cell migration and invasion were assessed using wound-healing and Transwell assay, and flow cytometry was used to analyze the cell cycle changes. The ultrastructural changes of the cells were observed by transmission electron microscopy., Results: Compared with the control group, the cells exposed to2 µmol/L AFB1 showed obviously enhanced migration and invasion with decreased cell ratio in G1/G1 phase and increased cell ratio in G2/M phase but no changes in S phase cells; transmission electron microscopy revealed the presence of multiple nucleoli and significantly increased mitochondria and Golgi apparatus in the exposed cells. Compared with AFB1-exposed cells, the cells treated with DHA and AFB1 showed decreased migration and invasion abilities, and the G1/G1 phase cells increased and G2/M phase cells decreased significantly; ultrastructurally, the cells contained single nucleoli with decreased mitochondria and vacuolization occurred in the cytoplasm., Conclusion: DHA can significantly inhibit AFB1-induced enhancement of cell migration and invasion in hepatocellular carcinoma cells in vitro.

The pig is an ideal source to provide organs because its organ size and physiology are similar to humans. However, an acute rejection will ensue after pig-to-human xenotransplantation. The α-1,3 galactosyltransferase gene knockout (GTKO) pigs were generated in recent years, and could solve the problem of hyperacute rejection. But due to lack of reporting genes, the rejection status of cells and organs post pig-to-human xenotransplantation cannot be visualized. In this study, we introduced the enhanced green fluorescent protein (EGFP) gene driven by the CAG promoter into GTKO porcine ear fibroblasts. Then we produced transgenic pigs expressing the EGFP gene by nuclear transfer technology. Expression levels of EGFP in different tissues and organs of the cloned pig were investigated by Nightsea DFP-1 Fluorescent Protein Flashlight, fluorescence microscope and quantitative PCR assays. The results showed that the protein and transcript of EGFP were expressed in all tissues and organs of the GTKO pig, but the expression was weak in the liver and central nervous system. In conclusion, we have successfully produced the transgenic GTKO pigs expressing EGFP in all tested tissues and organs, which builds up a good basis to track transplanted cells or tissues.

Objective: To identify the intergenotype recombination of hepatitis B virus (HBV) in Chinese patients and explore new recombination types of HBV., Methods: Complete genome sequences of HBV from Chinese patients were downloaded from GenBank and analyzed by Fragment-typing method to screen the potential recombinants, and the breakpoints of recombination were identified by using Simplot program., Results: Thirty-one recombination types including 755 recombinants were identified from 1642 complete HBV genome sequences, including 22 B/C recombination types with 676 recombinants, 5 C/D recombination types with 75 recombinants, 3 A/C recombination types with 3 recombinants, and 1 C/I recombination type with 1 recombinant., Conclusion: Of the 31 recombination types comprising 755 recombinants, 4 are novel recombination types found for the first time. All these recombination types identified in this study involve genotype C.

Disruption of myostatin (MSTN) gene in pigs may improve porcine lean meat percentage (LMP), and create an animal model for certain human diseases. Using zinc-finger nucleases (ZFNs) technology, MSTN gene was deleted in Wuzhishan miniature pig fibroblasts by transfection of either ZFNs plasmids or ZFNs mRNA in high efficiency. Strikingly, ZFNs encoding mRNA could produce MSTN+/-and MSTN-/- cell colonies with single or double allele deletion by single transfection. Sequencing results demonstrated that 92.18% of the mutations were short fragment deletions or insertions (≤10 bp). Prediction of amino acids sequences indicated that more than half of the mutations cause premature transla-tional-termination codon. MSTN+/+, MSTN+/-, and MSTN-/- cell colonies were used as nuclear donor for somatic cell nuclear transfer (SCNT), and developmental potential of SCNT embryos were measured by the blastocyst rate. The results revealed no significant difference in development competence among the three kinds of reconstructed embryos (14.29% vs. 19.64% vs. 16.13%), which provides the possibility of making myostatin knock out pigs in the future.

Objective: To systematically assess the clinical efficacy of bone morphogenetic proteins in the treatment of open tibial fractures., Methods: Based on the principles and methods of Cochrane systematic reviews, the Cochrane Library, PubMed, EMBASE, Chinese Bio-medicine Database, China Journal Full-text Database, VIP database were searched from their establishment to April 2012 in whatever language. Related journals were handsearched as well. Randomized controlled trials (RCTs) of bone morphogenetic protein for the treatment of open tibial fractures were included. The quality of the included trials according to the Cochrane Handbook for Systematic Reviews of Interventions Version was assessed. The Cochrane Collaboration's software RevMan 5.1 was used for meta-analysis., Results: Three RCTs totaling 851 patients were included. The results showed that bone morphogenetic protein had no significant differences in fracture healing [RR = 1.16, 95% CI (0.95,1.41), P = 0.15], but lower secondary interventions incidence rate [RR = 0.72, 95% CI (0.58, 0.89), P = 0.003]. There were no significant differences between the two groups in the incidence of adverse events of infection [RR = 1.31, 95% CI (0.94, 1.81), P = 0.11] and pain [RR = 0.92, 95% CI (0.79, 1.08), P = 0.30]., Conclusion: Bone morphogenetic protein has certain advantages in treating open tibial fractures. It needs more high-quality articles to assess the long-term effect of different courses of treatments. The above conclusion still needs more high-quality randomized controlled trails to be verified owing to the limitations of the number and quality of systematic review included studies.

The Hedgehog (Hh) signaling pathway inhibits fat accumulation, which is conserved in a wide variety of organisms from Drosophila to vertebrates, but few reports about its effect on pigs are available. In this study, pig Gli1 gene was cloned for the first time by rapid amplification of cDNA ends (RACE) and RT-PCR. Pig Gli1 expression profiles were then studied in different tissues and in different developmental stages of the adipose tissue of pigs using real-time PCR. Finally, the eukaryotic expression vector and the adipose tissue specific expression vector were constructed. The results showed that the full pig Gli1 cDNA length was 3 576 bp, the genomic sequence contained 10 715 bp with 12 exons, and 1 106 amino acids were encoded. Pig Gli1 was predicted as an unstable hydrophilic protein without a tans-membrane structure or a signal peptide. The C2H2 zinc finger domain and a nuclear localization sequence were found in pig Gli1. A homology analysis of the Gli1 amino acids and the genomic sequences among seven species showed that the identities were all greater than 80%, which indicates that Gli1 is highly conserved among different species. Tissue expression profile analysis showed that pig Gli1 was only expressed in the tone tissue of adult pigs. Analysis of the pig adipose tissue developmental process showed that Gli1 was detected in the adipose tissue of one-week-old pigs, but not in one-month-old and three-month-old pigs. Finally, a pig Gli1 eukaryotic expression vector was constructed and properly expressed with cell transfection. An adipose tissue specific expression vector was constructed for transgenic animal studies.

Objective: To investigate the effectiveness of reconstructing medial patellofemoral ligament with hamstring tendon autografts for the treatment of recurrent patellar dislocation under arthroscopy., Methods: Between January 2005 and January 2010, 22 cases of recurrent patellar dislocation were treated by lateral retinacular release and reconstruction of the medial patellofemoral ligament with hamstring tendon autografts under arthroscopy. There were 5 males and 17 females, aged 15-19 years (mean, 17.3 years). The average number of dislocation was 4 (range, 3-8). The main clinical symptoms were pain and swelling of knee joint, weakness in the leg, and limited range of motion (ROM). The patellar tilt test, pressing pain of patellofemoral ligament insertion, and apprehension sign showed positive results. According to International Knee Documentation Committee (IKDC) scoring criteria, the subjective IKDC score was 36.7 +/- 4.7, and the Lysholm score was 69.3 +/- 3.8. X-ray films showed that the patella inclined outwards., Results: All incisions healed by first intention. Twenty-two cases were followed up 18-49 months (mean, 34 months). Pain and swelling of knee joint and weakness were improved obviously. No recurrence was found during follow-up. The ROM of knee in flexion and extension was improved when compared with preoperative ROM. The subjective IKDC score was 92.4 +/- 5.3 and the Lysholm knee score was 91.7 +/- 5.2, showing significant differences when compared with preoperative scores (P < 0.05)., Conclusion: Reconstruction of the medial patellofemoral ligament with hamstring tendon autografts under arthroscopy is an effective method to treat recurrent patellar dislocation.

Objective: To investigate the clinical application of the reversed sural neurovascular fasciocutaneous flap., Methods: From January 2007 to May 2009,10 patients (6 males and 4 females) with soft tissue defects on lower limbs were treated by reversed sural neurovascular fasciocutaneous flap. The end-to-side neuroanastomosis were used to reinnervate the flap and microsurgery was used for anastomosing small saphenous vein. The age ranged from 16 to 55 years (mean, 32 years). There were 6 patient with soft tissue defects on foot, 2 patients with exposed bone after operation, 2 patients with chronic ulcer on limb. The patients were evaluated with appearance, blood supply, texture and 2-PD of the flaps., Results: All the patients were followed for 6 to 24 months (mean, 15 months). The flaps in all 10 patients survived completely. The appearance, blood supply and texture of the flaps were excellent and 2-PD was 9 to 12 mm., Conclusion: This flap has sufficient blood supply and high survival rate. It also effectively retains the feeling of dorsolateral heeland improved quality of life.

In the present study, the DNA methylation patterns of in vitro-derived mouse tetraploid embryos were investigated by immunofluorescence staining with an antibody against 5-methylcytosine (5MeC). Tetraploid embryos could be produced by electrofusion at the stage of two-cell embryos, which could develop to blastocysts followed by fusion of cytoplasm and nucleus and cleavage in vitro. During the fusion of cytoplasm, the DNA methylation levels of the fused embryos are as high as these of two-cell diploid embryos in vivo Then the embryos are rapidly demethylated when the nucleus begin to fuse, resulting in the lowest DNA methylation levels when the nucleus are fused completely. After that, the DNA methylation levels of the fused embryos are gradually increased until the morula stage. However, whereas an asymmetric distribution of DNA methylation is established in vivo-derived blastocysts with a higher methylation level in the inner cell mass (ICM) than that in the trophectoderm, we can not detect the asymmetric distribution in most in vitro-derived tetraploid blastocysts. So the DNA methylation patterns of mouse tetraploid embryos are aberrant, which may lead to subsequent developmental failure and embryo death. This is the first report on the methylation patterns of in vitro-derived mouse tetraploid embryos.

Objective: To investigate antibiotic resistance, clonal relatedness and carbapenemase genotype among carbapenem-resistant Acinetobacter baumannii collected from 3 comprehensive hospitals in Ningbo city, Zhejiang province., Methods: 28 strains of carbapenem resistant Acinetobacter baumannii were collected from Ningbo Li Hui-li Hospital, Ningbo Li Hui-li Hospital, Ningbo First Hospital, and N ingbo Second Hospital. The minimum inhibitory concentrations (MIC) of these strains were examined by agar dilution and E-test method. Homology of these isolates was analyzed by pulse-field gel electrophoresis (PFGE) and Genotype of carbapenemases were analyzed by PCR and verified by DNA sequencing., Results: 28 strains of Acinetobacter baumanii were highly resistant to all of the antibiotics except polymyxin E. They were classified into 4 clones based on PFGE pattern. Clone A and B had been spreading widely. All of the 28 strains produced carbapenemases which were confirmed as OXA-23 by PCR and sequencing. Metallo-beta-lactamase was not detected in any of the isolates., Conclusion: All of t hecarbapenem-resistant Acinetobacter baumannii collected from Ningbo were producing OXA-23 carbapenemase, suggesting that the transmission of clones had occurred in the 3 hospitals.

Objective: To study the clinical and pathological features of drug-induced liver injury (DILI)., Methods: Liver specimens were obtained through needle biopsies from 100 patients with DILI. The histological preparations of the specimens were stained with haematoxylin eosin, several histochemistry methods, and immunohistochemistry stains. The pathological changes of the livers were analyzed together with the patients's clinical data. The patients were divided into two groups, an acute DILI group (n=39) and a chronic DILI group (n=61), based on their clinical courses and histological changes in their livers. In the chronic DILI group, the clinical courses were longer than 6 months and/or fibrosis or cirrhosis occurred in their liver tissues., Results: Among our cases the leading cause of DILI was Chinese herb medicine, accounting for 21% of the 100 cases; steroids induced cases were 11% of the total. 78% of the patients presented elevated serum transaminases and/or jaundice. The degree of transaminases elevation and the frequency of jaundice happening in the acute group were significantly higher than those in the chronic group (P less than 0.05). The histopathological liver changes in these DILI cases included: (1) necrosis commonly occurred in acinar zone 3, (2) abundant neutrophil and/or eosinophil infiltrations, (3) hepatocytic and/or canalicular cholestasis with little or no inflammation, (4) microvesicular steatosis mixed with macrovesicular steatosis, and (5) presentation of epitheloid cell granuloma. There were no significant differences in liver histopathology between the acute and the chronic DILI groups, except that the fibrosis and the ductular proliferation were different., Conclusion: DILI has become a notable liver disease in mainland China, and the use of Chinese herbal medicine must be improved, standardized and regulated more closely.

Yunnan snub-nose monkey (Rhinopithecus bieti) is a famous endangered primate in China. So far, however, studies on its population genetics based on DNA sequences are not available. In this paper, the whole mitochondria control region of the samples from Weixi, Yunan Province as well as the whole cytochrome b gene in some individuals were sequenced. A deep divergence was observed within the Weixi population, which was confirmed after excluding the possibility of it being a nuclear pseudogene. Nonetheless, if the effects of population structure and migration are considered, the true level of polymorphism of the Weixi population may be not as high as observed.

Objective: To detect p24 antigen of human immunodeficiency virus (HIV)-1 in the liver biopsy specimens of patients with HIV infection., Methods: Liver biopsy samples from 14 patients with HIV/AIDS (11 man, 3 women; age range 27-52 years; infection time range 8-13 years) were examined by immunohistochemistry prospectively., Results: Intracellular expression of HIV-1 p24 antigen was detected in Kupffer cells, endothelial cells and hepatocytes. There were more HIV-positive liver cells in the patients with severer liver damage than those with milder liver damage (t=2.5189, P=0.0270)., Conclusion: These findings indicate that HIV-1 could replicate in the liver of HIV-infected patients and might be related to the liver cells apoptosis.

Background: To explore the pathological features and pathogenesis of severe acute respiratory syndrome (SARS) to provide evidence for the clinical treatment and prevention of SARS., Methods: Pathological features of 2 cases of full autopsy and 4 cases of needle biopsy tissue samples from the patients who died from SARS were studied by light and electron microscopy. The distribution and quantity of lymphocyte subpopulations in the lungs and immune organs from SARS patients were analyzed by immunohistochemistry. The location and semi-quantitative analysis of SARS coronavirus in the tissue specimens were studied by electron microscopy, in situ hybridization and immunohistochemistry., Results: In total of 6 cases, diffuse alveolar damage and alveolar cell proliferation were common. The major pathological changes of 2 autopsy cases of SARS in lung tissues were acute pulmonary interstitial and alveolar exudative inflammation, and 2 autopsy and one biopsy lung tissues showed alveolar hyaline membrane formation. Terminal bronchiolar and alveolar desquamation of lung tissues in one autopsy and 2 biopsy cases were noted. Among 6 cases, 2 biopsy cases presented early pulmonary fibrosis and alveolar organization. Meanwhile, the immune organs, including lymph nodes and spleens from 2 autopsy cases of SARS whose disease courses were less than 12 days showed extensive hemorrhagic necrosis, reactive macrophage/histocyte proliferation, with relative depression of mononuclear and granulocytic clones in the bone marrows. However, spleen and bone marrow biopsy tissue samples from 4 dead SARS cases whose clinical course lasted from 21 to 40 days presented repairing changes. SARS coronaviruses were mainly identified in type I and II alveolar epithelia, macrophages, and endothelia; meanwhile, some renal tubular epithelial cells, cardiomyocytes, mucosal and crypt epithelial cells of gastrointestinal tracts, parenchymal cells in adrenal glands, lymphocytes, testicular epithelial cells and Leydig's cells were also detected by electron microscopy combined with in situ hybridization. The semi-quantitative analysis of lymphocyte subpopulations revealed that the proportion of CD8+ T lymphocytes were about 80% of the total infiltrative inflammatory cells in the pulmonary interstitium, with a few CD4+ lymphocytes CD3+, CD4+, CD8+ or CD20+ lymphocyte subpopulations were obviously decreased and there was imbalance in number and proportion, while CD57+, CD68+, S-100+ and HLA-DR+ cells were relatively increased in lymph nodes and spleens., Conclusions: Histologically, the pulmonary changes could be divided into acute inflammatory exudative, terminal bronchiolar and alveolar desquamative and proliferative repair stages or types during the pathological process of SARS. SARS coronavirus was found in multi-target cells in vivo, which means that SARS coronavirus might cause multi-organ damages which were predominant in lungs. There were varying degrees of decrease and imbalance in number and proportion of lymphocyte subpopulations in the immune organs of the patients with SARS. However, these changes may be reversible. It was found that cellular immune responses were predominant in the lungs of SARS cases, which might play an important role in getting rid of coronaviruses in infected cells and inducing immune mediated injury.