Due to the pristine interface of the 2D/3D face-tunneling heterostructure with an ultra-sharp doping profile, the 2D/3D tunneling field-effect transistor(TFET) is considered as one of the most promising low-power devi...Due to the pristine interface of the 2D/3D face-tunneling heterostructure with an ultra-sharp doping profile, the 2D/3D tunneling field-effect transistor(TFET) is considered as one of the most promising low-power devices that can simultaneously obtain low off-state current(IOFF), high on-state current(ION) and steep subthreshold swing(SS). As a key element for the 2D/3D TFET, the intensive exploration of the tunnel diode based on the 2D/3D heterostructure is in urgent need.The transfer technique composed of the exfoliation and the release process is currently the most common approach to fabricating the 2D/3D heterostructures. However, the well-established transfer technique of the 2D materials is still unavailable.Only a small part of the irregular films can usually be obtained by mechanical exfoliation, while the choice of the chemical exfoliation may lead to the contamination of the 2D material films by the ions in the chemical etchants. Moreover, the deformation of the 2D material in the transfer process due to its soft nature also leads to the nonuniformity of the transferred film,which is one of the main reasons for the presence of the wrinkles and the stacks in the transferred film. Thus, the large-scale fabrication of the high-quality 2D/3D tunnel diodes is limited. In this article, a comprehensive transfer technique that can mend up the shortages mentioned above with the aid of the water and the thermal release tape(TRT) is proposed. Based on the method we proposed, the MoS_(2)/Si tunnel diode is experimentally demonstrated and the transferred monolayer MoS_(2) film with the relatively high crystal quality is confirmed by atomic force microscopy(AFM), scanning electron microscopy(SEM), and Raman characterizations. Besides, the prominent negative differential resistance(NDR) effect is observed at room temperature, which verifies the relatively high quality of the MoS_(2)/Si heterojunction. The bilayer MoS_(2)/Si tunnel diode is also experimentally fabricated by repeating the transfer process we proposed, followed by the specific analysis of the electrical characteristics. This study shows the advantages of the transfer technique we proposed and indicates the great application foreground of the fabricated 2D/3D heterostructure for ultralow-power tunneling devices.展开更多
AIM: To investigate the ability of a genetically altered embryonic stem (ES) cell line to generate insulin-producing cells in vitro following transfer of the Nkx2.2 gene.METHODS: Hamster Nkx2.2 genes were transfer...AIM: To investigate the ability of a genetically altered embryonic stem (ES) cell line to generate insulin-producing cells in vitro following transfer of the Nkx2.2 gene.METHODS: Hamster Nkx2.2 genes were transferred into mouse ES cells. Parental and Nkx2.2-transfected ES cells were initiated toward differentiation in embryoid body (EB) culture for 5 d and the resulting EBs were transferred to an attached culture system. Dithizone (DTZ), a zincchelating agent known to selectively stain pancreatic beta cells, was used to detect insulin-producing cells.The outgrowths were incubated in DTZ solution (final concentration, 100μg/mL) for 15 rain before being examined microscopically. Gene expression of the endocrine pancreatic markers was also analyzed by RT-PCR. In addition, insulin production was determined immunohistochemically and its secretion was examined using an ELISA.RESULTS: DTZ-stained cellular clusters appeared after approximately 14 d in the culture of Nkx2.2-transfected ES cells (Nkx-ES cells), which was as much as 2 wk earlier, than those in the culture of parental ES cells (wt-ES). The frequency of DTZ-positive cells among total cultured cells on day 28 accounted for approximately 1.0% and 0.1% of the Nkx-ES- and wt-ES-derived EB outgrowths, respectively. The DTZ-positive cellular clusters were found to be immunoreactive to insulin, while the gene expressions of pancreatic-duodenal homeobox 1 (PDX1), proinsulin 1 and proinsulin 2 were observed in the cultures that contained DTZ-positive cellular clusters.Insulin secretion was also confirmed by ELISA, whereas glucose-dependent secretion was not demonstrated.CONCLUSION: Nkx2.2-transfected ES cells showed an ability to differentiate into insulin-producing cells.展开更多
Deep-sea sediment is extremely important in marine scientific research,such as that concerning marine geology and microbial communities.The research findings are closely related to the in-situ information of the sedim...Deep-sea sediment is extremely important in marine scientific research,such as that concerning marine geology and microbial communities.The research findings are closely related to the in-situ information of the sediment.One prerequisite for investigations of deep-sea sediment is providing sampling techniques capable of preventing distortion during recovery.As the fruit of such sampling techniques,samplers designed for obtaining sediment have become indispensable equipment,owing to their low cost,light weight,compactness,easy operation,and high adaptability to sea conditions.This paper introduces the research and application of typical deep-sea sediment samplers.Then,a representative sampler recently developed in China is analyzed.On this basis,a review and analysis is conducted regarding the key techniques of various deep-sea sediment samplers,including sealing,pressure and temperature retaining,low-disturbance sampling,and no-pressure drop transfer.Then,the shortcomings in the key techniques for deep-sea sediment sampling are identified.Finally,prospects for the future development of key techniques for deep-sea sediment sampling are proposed,from the perspectives of structural diversification,functional integration,intelligent operation,and high-fidelity samples.This paper summarizes the existing samplers in the context of the key techniques mentioned above,and can provide reference for the optimized design of samplers and development of key sampling techniques.展开更多
Aim: To investigate the effects of rat Erythropoietin (Epo) on spermatogenesis by transferring rat Epo gene into cryptorchid testes by means of in vivo electroporation. Methods: Sprague-Dawley rats with surgically...Aim: To investigate the effects of rat Erythropoietin (Epo) on spermatogenesis by transferring rat Epo gene into cryptorchid testes by means of in vivo electroporation. Methods: Sprague-Dawley rats with surgically-induced unilateral cryptorchidism were divided into three groups: the first group was given intratesticular injections of pCAGGS-Epo (pCAGGS-Epo group), the second group was given intratesticular injections of pCAGGS (pCAGGS group), and the third group were given intratesticular injections of phosphate-buffered saline (PBS group). At the same time, square electric pulses of 30 V were applied six times with a time constant of 100 ms. One or two weeks after injection, each testis was weighed and the ratio of the total number of germ cells to that of Sertoli cells (G/S ratio) was calculated to evaluate the impairment of spermatogenesis. Ten testes taken from each of the three groups were examined at each time point. Results: The testicular weight after the injection of pCAGGS-Epo or pCAGGS control plasmid was (0.85 ± 0.08) g and (0.83 ± 0.03) g, respectively, at week 1 (P = 0.788) and (0.62 ± 0.06) g and (0.52 ± 0.02) g, respectively, at week 2 (P = 0.047). At week 1, spermatids and sperm were more abundant in testes with pCAGGS-Epo than those in the control testes. At week 2, spermatids and sperm were hardly detected in either group. The G/S ratio was 23.27 ± 6.80 vs. 18.63 ± 5.30 at week 1 (P = 0.0078) and 7.16 ± 3.06 vs. 6.05 ± 1.58 at week 2 (P = 0.1471), respectively. Conclusion: The transfer of Epo to rat testes by in vivo electroporation may reduce the risk of the germ cell loss caused by cryptorchidism.展开更多
To examine the ability of intrastriatal gene transfer of vascular endothelial growth factor 165 mediated by adenoviral vector to rescue dopaminergic neurons in a rat model of Parkinson's disease (PD), we constructe...To examine the ability of intrastriatal gene transfer of vascular endothelial growth factor 165 mediated by adenoviral vector to rescue dopaminergic neurons in a rat model of Parkinson's disease (PD), we constructed recombinant replication-deficent adenoviral vectors carrying the gene of VEGF165 (Ad-VEGF), and injected Ad-VEGF (or Ad-LacZ and PBS as controls) into the striatum of rats 7 days after the lesion by 6-hydroxydopamine. The rat rotational behavior analysis and tyrosine hydroxylase (TH) immunohistochemistry were performed to assess the change of dopaminergic neurons. Our results showed that the rats receiving Ad-VEGF injection displayed a significant improvement in apomorphine-induced rotational behavior and a significant preservation of TH-positive neurons and fibers compared with control animals, It is concluded that intrastriatal gene transfer by Ad-VEGF may rescue the dopaminergic neurons from degeneration in a rat model of PD.展开更多
[ Objective] The paper was to test the feasibility of embryo transfer technique in pig production. [ Method ] Twenty-four estrus muhiparity Landraee sows provided by Longjing Agricultural Science and Technology Instit...[ Objective] The paper was to test the feasibility of embryo transfer technique in pig production. [ Method ] Twenty-four estrus muhiparity Landraee sows provided by Longjing Agricultural Science and Technology Institute were performed embryo transfer surgery, and postoperative effects were observed. [ Result] Totally 11 out of 24 receptor sows were pregnant ; pregnant sows delivered 67 cloned piglets, and the average farrowing rate of sows was approximately 6 piglets/sow. There were 22 mortalities of newborn piglets because of various reasons. [ Conclusion] Embryo transfer technique is an indispensable link in pig production, and an important means of pig breeding and improvement.展开更多
Oxygen transfer presents a serious challenge in the application of liquid lead as a nuclear coolant in advanced reactors. To mitigate corrosion by liquid lead in contact with steel, carefully controlling the oxygen co...Oxygen transfer presents a serious challenge in the application of liquid lead as a nuclear coolant in advanced reactors. To mitigate corrosion by liquid lead in contact with steel, carefully controlling the oxygen concentration has been used as an effective way. Oxygen needs to mix in liquid lead uniformly and quickly. To enhance oxygen transport in liquid lead, nanoparticles are added to the liquid metal. In the current study, a lattice Boltzmann method is applied to investigate natural convection of copper/lead and aluminum oxide/lead in two-dimensional simplified container. Two thermal boundary cases are evaluated in order to check the effect of different natural convection flow patterns on oxygen transport. Some useful information are obtained such as improvement in natural convection and reduction in oxygen equilibrium time.展开更多
Face authentication is an important biometric authentication method commonly used in security applications.It is vulnerable to different types of attacks that use authorized users’facial images and videos captured fr...Face authentication is an important biometric authentication method commonly used in security applications.It is vulnerable to different types of attacks that use authorized users’facial images and videos captured from social media to perform spoofing attacks and dynamic movements for penetrating secur-ity applications.This paper presents an innovative challenge-response emotions authentication model based on the horizontal ensemble technique.The proposed model provides high accurate face authentication process by challenging the authorized user using a random sequence of emotions to provide a specific response for every authentication trial with a different sequence of emotions.The proposed model is applied to the KDEF dataset using 10-fold cross-valida-tions.Several improvements are made to the proposed model.First,the VGG16 model is applied to the seven common emotions.Second,the system usability is enhanced by analyzing and selecting only the four common and easy-to-use emotions.Third,the horizontal ensemble technique is applied to enhance the emotion recognition accuracy and minimize the error during authen-tication processes.Finally,the Horizontal Ensemble Best N-Losses(HEBNL)is applied using challenge-response emotion to improve the authentication effi-ciency and minimize the computational power.The successive improvements implemented on the proposed model led to an improvement in the accuracy from 92.1%to 99.27%.展开更多
Gastric cancer is common in China [1-42],and its early diagnosis and treatment in advanced stage are difficult [31-50].In recent years ,gene study in cancer is a hotspot ,and great progress has been achieved [41-80] ....Gastric cancer is common in China [1-42],and its early diagnosis and treatment in advanced stage are difficult [31-50].In recent years ,gene study in cancer is a hotspot ,and great progress has been achieved [41-80] .Cancer gene therapy has shifted from the imagination into the laboratory and clinical trials.展开更多
Aim: To study polyethylenimine (PEI)-mediated in vivo gene transfection into testis cells and preliminary functional research of spermatogenic cell-specific gene NYD-SP12 using this method. Methods: PEI/DNA comple...Aim: To study polyethylenimine (PEI)-mediated in vivo gene transfection into testis cells and preliminary functional research of spermatogenic cell-specific gene NYD-SP12 using this method. Methods: PEI/DNA complexes were introduced into the seminiferous tubules of mouse testes using intratesticular injection. Transfection efficiency and speciality were analyzed on the third day of transfection with fluorescent microscopy and hematoxylin staining. The long-lasting expression of the GFP-NYD-SP12 fusion protein and its subcelluar localization in spermatogenic cells at different stages were analyzed with fluorescent microscopy and propidium iodide staining. Results: With the mediation of PEI, the GFP-NYD-SP12 fusion gene was efficiently transferred and expressed in the germ cells (especially in primary spermatocytes). Transfection into Sertoli cells was not observed. The subcellular localization of the GFP-NYD-SP2 fusion protein showed dynamic shifts in spermatogenic cells at different stages during spermatogenesis. Conclusion: PEI can efficiently mediate gene transfer into spermatocytes. Thus, it might be useful for the functional research of spermatogenic-cell specific genes such as the NYD-SP12 gene. In our gtudy, the NYD-SP12 protein was visualized and was involved in the formation of acrosome during spermatogenesis. Our research will continue into the detailed function of NYD-SP12 in spermatocytes. (Asian J Androl 2006 Jan; 8: 53-59)展开更多
The topside floatover installation is always a great challenge and is sensitive to environmental conditions.In this study,experimental analysis on the mating operation of the floatover installation in different wave h...The topside floatover installation is always a great challenge and is sensitive to environmental conditions.In this study,experimental analysis on the mating operation of the floatover installation in different wave headings is presented.The continuous mating operation using the rapid transfer technique was experimentally simulated with the assistance of the jacking system and the ballast system.In the continuous transfer modeling,the topsides loads were transferred onto the jacket by several consecutive steps,including the first rapid jack-down for the 30%loads,continuous 30%−70%load transfer and the second repaid jack-down for the remaining 30%loads.Motions of the barge and the topsides as well as loads on the Deck Support Unite(DSU)and the Leg Mating Unite(LMU)in different wave headings were measured.Experimental results illustrated the complex motion behavior and load characteristics of the continuous transfer operation.Results indicate that the rapid jack-down operations will lead to impact loads and larger lateral DSU loads.The bow quartering seas are much more dangerous as it gives rise to the larger motions and loads.Comparisons with the traditional steady-state modeling indicate that the continuous transfer modeling has greater advantages over the steady-state modeling on predicting the loads.展开更多
Monolayer transition-metal dichalcogenides (TMDs) are considered to be fantastic building blocks for a wide variety of optical and optoelectronic devices such as sensors, photodetectors, and quantum emitters, owing ...Monolayer transition-metal dichalcogenides (TMDs) are considered to be fantastic building blocks for a wide variety of optical and optoelectronic devices such as sensors, photodetectors, and quantum emitters, owing to their direct band gap, transparency, and mechanical flexibility. The core element of many conventional electronic and optoelectronic devices is the p-n junction, in which the p- and n-types of the semiconductor are formed by chemical doping in different regions. Here, we report a series of optoelectronic studies on a monolayer WSe2 in-plane p-n photodetector, demonstrating a low- power dissipation by showing an ambipolar behavior with a reduced threshold voltage by a factor of two compared with the previous results on a lateral electrostatically doped WSe2 p-n junction. The fabrication of the device is based on a polycarbonates (PC) transfer technique and hence no electron-beam exposure induced damage to the monolayer WSe2 is expected. Upon optical excitation, the photodetector demonstrates a photoresponsivity of 0.12 mA.W-1 and a maximum external quantum efficiency of 0.03%. Our study provides an alternative platform for a flexible and transparent two- dimensional photodetector, from which we expect to further promote the development of next-generation optoelectronic devices.展开更多
The gene encoding nonstructural protein Npro of BVDV was amplified by PCR,then was inserted into pEGFP-C1 vector.The 293 cells were transfected in vitro with recombinant plasmid by liposome.Npro-EGFP fusion protein wa...The gene encoding nonstructural protein Npro of BVDV was amplified by PCR,then was inserted into pEGFP-C1 vector.The 293 cells were transfected in vitro with recombinant plasmid by liposome.Npro-EGFP fusion protein was viewed directly with fluorensce microscope and mRNA expression of Npro was detected by reverse transcription-polymerase chain reaction(RT-PCR).Results showed that recombinant plasmid was confirmed to be constructed correctly by PCR,restriction enzyme digestion and DNA sequencing;meanwhile,the gene carried was expressed in 293 cells.The fusion protein had properties of both the two Npro and enhanced green fluorescent proteins(EGFP) and distributed in the cytoplasm.This research lays a foundation for further researches which explored Npro gene function in the process of viral replication and synthesis.展开更多
Parkinson’s disease patients suffer from disorders of speech.The most frequently reported speech problems are weak,hoarse,nasal or monotonous voice,imprecise articulation,slow or fast speech,difficulty starting speec...Parkinson’s disease patients suffer from disorders of speech.The most frequently reported speech problems are weak,hoarse,nasal or monotonous voice,imprecise articulation,slow or fast speech,difficulty starting speech,impaired stress or rhythm,stuttering,and tremor.To improve the speech quality and assist the patient with speech rehabilitation therapy,we have proposed the speech recognition model for Parkinson’s disease patients using transfer learning technique(PSTL),where we have pre-trained the long short-term memory(LSTM)neural network model with our developed publicly available dataset that has been obtained from healthy people through the social media platform.Then,we applied the transfer learning technique to improve the performance of the PSTL framework.The frequency spectrogram masking data augmentation method has been used to alleviate the over-fitting problem so that the word error rate(WER)is further reduced.Even with a limited dataset,our proposed model has effectively reduced the WER from 58% to 44.5% on the original speech dataset and 53.1% to 43% on the denoised speech dataset,which demonstrated the feasibility of our framework.展开更多
Extracts of elephant grass (Penniseturn purpureum) blended with some intensifier halides like ammonium chloride (AMC) and potassium iodide (PTI) were investigated as corrosion inhibitor for mild steel. The corro...Extracts of elephant grass (Penniseturn purpureum) blended with some intensifier halides like ammonium chloride (AMC) and potassium iodide (PTI) were investigated as corrosion inhibitor for mild steel. The corrosion process was monitored in 3.5% HCI by mass loss and electrochemical techniques at 30, 40, 50, 60 and 90 ℃. Addition of AMC and PTI increased the inhibition efficiency with the highest inhibition efficiency obtained with PTI blend- ed extract. The blends behaved as mixed type inhibitors and were spontaneously adsorbed on mild steel surface in exothermic nature. Synergistic parameters of the intensifier ions revealed cooperative effect. Kinetic data treatment indicated increase in energy barrier by intensifier ions. The results demonstrate that elephant grass extract blended with halide ions can act as alternative ecofriendly inhibitor for mild steel at elevated temperatures.展开更多
OBJECTIVE: To investigate the feasibility and features of 3 catheter-mediated approaches of gene transfer into heart, including direct myocardial injection (DMI), coronary artery perfusion (CAP), and intrapericardial ...OBJECTIVE: To investigate the feasibility and features of 3 catheter-mediated approaches of gene transfer into heart, including direct myocardial injection (DMI), coronary artery perfusion (CAP), and intrapericardial cavity injection (ICI). METHODS: Fifteen dogs were used, and 0.3 ml (1 x 10(9) pfu) of an adenovirus (Adex1SR LacZ) was injected into the heart by 3 methods. The dogs were killed 5 days following injection, and gene expressions in heart and liver were evaluated by histochemical analysis. RESULTS: The results showed that (1) the CAP method was relatively less damaging and induced sparse LacZ expression in the myocardium, and the gene expression was also found in both vessels within the myocardium and liver; (2) gene transfer by DMI resulted in intense LacZ expression around the injection accompanied by a local inflammatory response; (3) LacZ expression elicited by ICI was detected in either the inner surface of the parietal pericardium or epicardial surface of the heart, and also in the myocardium underlying the visceral pericardium. CONCLUSION: Three catheter-mediated methods of gene transfer into the heart may be used and a reasonable approach should be chosen according to purpose.展开更多
OBJECTIVE: To determine the efficacy of adenovirus mediated suicide gene transduction combined with prodrug 5-fluorocytosine (5FC) as a therapeutic protocol for pancreatic cancer. METHODS: Cytosine Deaminase(CD) gene...OBJECTIVE: To determine the efficacy of adenovirus mediated suicide gene transduction combined with prodrug 5-fluorocytosine (5FC) as a therapeutic protocol for pancreatic cancer. METHODS: Cytosine Deaminase(CD) gene was cloned into pAdTrack-CMV-CD, pAdTrack-CMV-CD and pAdEasy-1 were recombined in bacteria. The newly recombined adenovirus (Ad)-CD containing green fluorescent protein (GFP) were packaged and propagated in 293 cells and purified by cesium chloride gradient centrifugation. Human pancreatic carcinoma cell line-Patu8988 was infected with this virus, then 5FC was added. XTT assay was used to estimate relative numbers of viable cells. In vivo model of pancreatic cancer was established by injecting 1.0 x 10(7) Patu8988 cells subcutaneously in Balb/c nude mice. When tumors were palpable, Ad-CD was injected into each tumor and 5FC was administered. RESULTS: Positive clones were selected using endonuclease to digest the recombinants and the concentration of viral liquids containing the CD gene was 2 x 10(11) pfu /ml. Significant cytotoxic activity as shown for 5FC in the CD gene transduced 8988 cell line, while little effect was found in the nontransduced pancreatic carcinoma cells. Antitumor effect was observed in Patu8988 xenograft nude mice with in situ CD gene transduction. CONCLUSIONS: CD gene mediated by adenovirus has high infectivity and may be useful for gene therapy in pancreatic carcinoma. These data demonstrate the use of an enzyme prodrug strategy in experimental pancreatic cancer.展开更多
OBJECTIVE: To investigate the feasibility of endothelialization of bioprosthesis by transfer of vascular endothelial growth factor (VEGF) gene. METHODS: Bovine pericardium treated with glutaraldehyde and L-glutamic ac...OBJECTIVE: To investigate the feasibility of endothelialization of bioprosthesis by transfer of vascular endothelial growth factor (VEGF) gene. METHODS: Bovine pericardium treated with glutaraldehyde and L-glutamic acid was positioned into the pig right atrium. pcD(2)/hVEGF(121) gene (1 mg) was transferred into the right ventricular myocardium using surgical sutures Reverse transcri ption polymerase chain reaction (RT PCR) was employed to evaluate the expression of myocardial VEGF mRNA. The determination of concentrations of VEGF protein in blood from both the right atrium and peripheral vein, and histological and ultrastructural analysis of implanted bovine pericardium were completed simultaneously. RESULTS: The concentration of VEGF derived from the right atrium in pcD(2)/hVEGF(121) group was significantly higher than that in the pcD(2) group 10 days after VEGF gene transfer (P展开更多
OBJECTIVE: To explore the inhibitory effect of human endostatin gene mediated by retroviral vector on the growth of human liver carcinoma. METHODS: A recombinant retroviral plasmid containing human endostatin gene and...OBJECTIVE: To explore the inhibitory effect of human endostatin gene mediated by retroviral vector on the growth of human liver carcinoma. METHODS: A recombinant retroviral plasmid containing human endostatin gene and signal peptide was engineered and transferred into PA317 cells to produce retrovirus. Human liver carcinoma cells (SMMC7721) were infected with the above retrovirus to build a stable endostatin-transfected liver carcinoma cell line (SMMC-endo). The control liver carcinoma cell line (SMMC-pLncx) was developed in a similar way except that the plasmid was replaced by an empty retroviral vector. Immunohistochemistry and Western blot were used to test the expression and secretion of human endostatin. The biological activity of the expressed human endostatin was assessed by endothelial cell proliferation assay. The growth rates of SMMC-endo and control SMMC-pLncx cells in vivo and in vitro were also observed. RESULTS: The expression and secretion of human endostatin by endostatin-transfected SMMC-endo cells were confirmed by immunohistochemistry and Western blot. Compared with the control group, concentrated supernatant of SMMC-endo cells remarkably inhibited the proliferation of human umbilical vein endothelial cells by 48%, significantly higher than the inhibition by the control (10.2%; P展开更多
OBJECTIVE: To specifically deliver the therapeutic gene to cancer cells and construct target retroviral vectors by inserting the single-chain variable antibody fragment into the retroviral envelope. METHODS: Single-ch...OBJECTIVE: To specifically deliver the therapeutic gene to cancer cells and construct target retroviral vectors by inserting the single-chain variable antibody fragment into the retroviral envelope. METHODS: Single-chain antibody expression vector pET -20bScfv was constructed. Binding activity of the genetically engineered single-chain variable antibody fragment was verified by enzyme-linked immunosorbent assay (ELISA) and Western blot. At the same time, by means of polymerase chain reaction (PCR)-directed mutagenesis, the appropriate cloning site EcoT22/Sal was generated at the N-terminus of receptor-binding SU domain in the MoMLV env polypeptide. Then the single- chain antibody gene, encoding a functional antibody, was inserted into the cloning site. The Scfv-env fusion gene fragment was subcloned into CMV expression vector. The Lac-Z retrovirus that co-displayed the Scfv-env chimeric protein and wild-type env protein was produced by transfection of Psi 2 cells with retroviral plasmid and the fusion gene expressing plasmid.To confirm the specificity of the recombinant retrovirus, infection assays and competitive inhibition assays were performed. RESULTS: The results of ELISA and Western blot showed that the genetically engineered single-chain variable antibody fragment could bind to the SHG44 cells surface membrane antigen. Virus-binding assay, viral infection and competitive inhibition assays confirmed that the harvested virus efficiently bound to and infected SHG44 cancer cells expressing the relative membrane antigen specially via the recognition of the target antigen. CONCLUSION: These results imply that insertion of Scfv into the retroviral envelope can specifically deliver the interested gene into specific antigen-producing cancer cells.展开更多
基金Project supported by the National Natural Science Foundation of China (Grant No.61851405)。
文摘Due to the pristine interface of the 2D/3D face-tunneling heterostructure with an ultra-sharp doping profile, the 2D/3D tunneling field-effect transistor(TFET) is considered as one of the most promising low-power devices that can simultaneously obtain low off-state current(IOFF), high on-state current(ION) and steep subthreshold swing(SS). As a key element for the 2D/3D TFET, the intensive exploration of the tunnel diode based on the 2D/3D heterostructure is in urgent need.The transfer technique composed of the exfoliation and the release process is currently the most common approach to fabricating the 2D/3D heterostructures. However, the well-established transfer technique of the 2D materials is still unavailable.Only a small part of the irregular films can usually be obtained by mechanical exfoliation, while the choice of the chemical exfoliation may lead to the contamination of the 2D material films by the ions in the chemical etchants. Moreover, the deformation of the 2D material in the transfer process due to its soft nature also leads to the nonuniformity of the transferred film,which is one of the main reasons for the presence of the wrinkles and the stacks in the transferred film. Thus, the large-scale fabrication of the high-quality 2D/3D tunnel diodes is limited. In this article, a comprehensive transfer technique that can mend up the shortages mentioned above with the aid of the water and the thermal release tape(TRT) is proposed. Based on the method we proposed, the MoS_(2)/Si tunnel diode is experimentally demonstrated and the transferred monolayer MoS_(2) film with the relatively high crystal quality is confirmed by atomic force microscopy(AFM), scanning electron microscopy(SEM), and Raman characterizations. Besides, the prominent negative differential resistance(NDR) effect is observed at room temperature, which verifies the relatively high quality of the MoS_(2)/Si heterojunction. The bilayer MoS_(2)/Si tunnel diode is also experimentally fabricated by repeating the transfer process we proposed, followed by the specific analysis of the electrical characteristics. This study shows the advantages of the transfer technique we proposed and indicates the great application foreground of the fabricated 2D/3D heterostructure for ultralow-power tunneling devices.
文摘AIM: To investigate the ability of a genetically altered embryonic stem (ES) cell line to generate insulin-producing cells in vitro following transfer of the Nkx2.2 gene.METHODS: Hamster Nkx2.2 genes were transferred into mouse ES cells. Parental and Nkx2.2-transfected ES cells were initiated toward differentiation in embryoid body (EB) culture for 5 d and the resulting EBs were transferred to an attached culture system. Dithizone (DTZ), a zincchelating agent known to selectively stain pancreatic beta cells, was used to detect insulin-producing cells.The outgrowths were incubated in DTZ solution (final concentration, 100μg/mL) for 15 rain before being examined microscopically. Gene expression of the endocrine pancreatic markers was also analyzed by RT-PCR. In addition, insulin production was determined immunohistochemically and its secretion was examined using an ELISA.RESULTS: DTZ-stained cellular clusters appeared after approximately 14 d in the culture of Nkx2.2-transfected ES cells (Nkx-ES cells), which was as much as 2 wk earlier, than those in the culture of parental ES cells (wt-ES). The frequency of DTZ-positive cells among total cultured cells on day 28 accounted for approximately 1.0% and 0.1% of the Nkx-ES- and wt-ES-derived EB outgrowths, respectively. The DTZ-positive cellular clusters were found to be immunoreactive to insulin, while the gene expressions of pancreatic-duodenal homeobox 1 (PDX1), proinsulin 1 and proinsulin 2 were observed in the cultures that contained DTZ-positive cellular clusters.Insulin secretion was also confirmed by ELISA, whereas glucose-dependent secretion was not demonstrated.CONCLUSION: Nkx2.2-transfected ES cells showed an ability to differentiate into insulin-producing cells.
基金Supported by National Key Research and Development Program of China(Grant No.2016YFC0300502)Hunan Provincial Innovation Foundation For Postgraduate(Grant No.CX2018B658)+2 种基金National Natural Science Foundation of China(Grant Nos.51705145,517779092)Supported by Scientific Research Fund of Hunan Provincial Education Department(Grant No.18B205)Hunan Province Natural Science Foundation(Grant No.2019 JJ50182).
文摘Deep-sea sediment is extremely important in marine scientific research,such as that concerning marine geology and microbial communities.The research findings are closely related to the in-situ information of the sediment.One prerequisite for investigations of deep-sea sediment is providing sampling techniques capable of preventing distortion during recovery.As the fruit of such sampling techniques,samplers designed for obtaining sediment have become indispensable equipment,owing to their low cost,light weight,compactness,easy operation,and high adaptability to sea conditions.This paper introduces the research and application of typical deep-sea sediment samplers.Then,a representative sampler recently developed in China is analyzed.On this basis,a review and analysis is conducted regarding the key techniques of various deep-sea sediment samplers,including sealing,pressure and temperature retaining,low-disturbance sampling,and no-pressure drop transfer.Then,the shortcomings in the key techniques for deep-sea sediment sampling are identified.Finally,prospects for the future development of key techniques for deep-sea sediment sampling are proposed,from the perspectives of structural diversification,functional integration,intelligent operation,and high-fidelity samples.This paper summarizes the existing samplers in the context of the key techniques mentioned above,and can provide reference for the optimized design of samplers and development of key sampling techniques.
文摘Aim: To investigate the effects of rat Erythropoietin (Epo) on spermatogenesis by transferring rat Epo gene into cryptorchid testes by means of in vivo electroporation. Methods: Sprague-Dawley rats with surgically-induced unilateral cryptorchidism were divided into three groups: the first group was given intratesticular injections of pCAGGS-Epo (pCAGGS-Epo group), the second group was given intratesticular injections of pCAGGS (pCAGGS group), and the third group were given intratesticular injections of phosphate-buffered saline (PBS group). At the same time, square electric pulses of 30 V were applied six times with a time constant of 100 ms. One or two weeks after injection, each testis was weighed and the ratio of the total number of germ cells to that of Sertoli cells (G/S ratio) was calculated to evaluate the impairment of spermatogenesis. Ten testes taken from each of the three groups were examined at each time point. Results: The testicular weight after the injection of pCAGGS-Epo or pCAGGS control plasmid was (0.85 ± 0.08) g and (0.83 ± 0.03) g, respectively, at week 1 (P = 0.788) and (0.62 ± 0.06) g and (0.52 ± 0.02) g, respectively, at week 2 (P = 0.047). At week 1, spermatids and sperm were more abundant in testes with pCAGGS-Epo than those in the control testes. At week 2, spermatids and sperm were hardly detected in either group. The G/S ratio was 23.27 ± 6.80 vs. 18.63 ± 5.30 at week 1 (P = 0.0078) and 7.16 ± 3.06 vs. 6.05 ± 1.58 at week 2 (P = 0.1471), respectively. Conclusion: The transfer of Epo to rat testes by in vivo electroporation may reduce the risk of the germ cell loss caused by cryptorchidism.
文摘To examine the ability of intrastriatal gene transfer of vascular endothelial growth factor 165 mediated by adenoviral vector to rescue dopaminergic neurons in a rat model of Parkinson's disease (PD), we constructed recombinant replication-deficent adenoviral vectors carrying the gene of VEGF165 (Ad-VEGF), and injected Ad-VEGF (or Ad-LacZ and PBS as controls) into the striatum of rats 7 days after the lesion by 6-hydroxydopamine. The rat rotational behavior analysis and tyrosine hydroxylase (TH) immunohistochemistry were performed to assess the change of dopaminergic neurons. Our results showed that the rats receiving Ad-VEGF injection displayed a significant improvement in apomorphine-induced rotational behavior and a significant preservation of TH-positive neurons and fibers compared with control animals, It is concluded that intrastriatal gene transfer by Ad-VEGF may rescue the dopaminergic neurons from degeneration in a rat model of PD.
基金Supported by National Transgenic Major Project Fund(2009ZX08006-002B)
文摘[ Objective] The paper was to test the feasibility of embryo transfer technique in pig production. [ Method ] Twenty-four estrus muhiparity Landraee sows provided by Longjing Agricultural Science and Technology Institute were performed embryo transfer surgery, and postoperative effects were observed. [ Result] Totally 11 out of 24 receptor sows were pregnant ; pregnant sows delivered 67 cloned piglets, and the average farrowing rate of sows was approximately 6 piglets/sow. There were 22 mortalities of newborn piglets because of various reasons. [ Conclusion] Embryo transfer technique is an indispensable link in pig production, and an important means of pig breeding and improvement.
基金the financial support from the office of Vice-President for Research at University of Nevada-Las Vegas
文摘Oxygen transfer presents a serious challenge in the application of liquid lead as a nuclear coolant in advanced reactors. To mitigate corrosion by liquid lead in contact with steel, carefully controlling the oxygen concentration has been used as an effective way. Oxygen needs to mix in liquid lead uniformly and quickly. To enhance oxygen transport in liquid lead, nanoparticles are added to the liquid metal. In the current study, a lattice Boltzmann method is applied to investigate natural convection of copper/lead and aluminum oxide/lead in two-dimensional simplified container. Two thermal boundary cases are evaluated in order to check the effect of different natural convection flow patterns on oxygen transport. Some useful information are obtained such as improvement in natural convection and reduction in oxygen equilibrium time.
基金This work is partially supported by the Deanship of Scientific Research at Jouf University under Grant No(DSR-2021–02–0369).
文摘Face authentication is an important biometric authentication method commonly used in security applications.It is vulnerable to different types of attacks that use authorized users’facial images and videos captured from social media to perform spoofing attacks and dynamic movements for penetrating secur-ity applications.This paper presents an innovative challenge-response emotions authentication model based on the horizontal ensemble technique.The proposed model provides high accurate face authentication process by challenging the authorized user using a random sequence of emotions to provide a specific response for every authentication trial with a different sequence of emotions.The proposed model is applied to the KDEF dataset using 10-fold cross-valida-tions.Several improvements are made to the proposed model.First,the VGG16 model is applied to the seven common emotions.Second,the system usability is enhanced by analyzing and selecting only the four common and easy-to-use emotions.Third,the horizontal ensemble technique is applied to enhance the emotion recognition accuracy and minimize the error during authen-tication processes.Finally,the Horizontal Ensemble Best N-Losses(HEBNL)is applied using challenge-response emotion to improve the authentication effi-ciency and minimize the computational power.The successive improvements implemented on the proposed model led to an improvement in the accuracy from 92.1%to 99.27%.
文摘Gastric cancer is common in China [1-42],and its early diagnosis and treatment in advanced stage are difficult [31-50].In recent years ,gene study in cancer is a hotspot ,and great progress has been achieved [41-80] .Cancer gene therapy has shifted from the imagination into the laboratory and clinical trials.
文摘Aim: To study polyethylenimine (PEI)-mediated in vivo gene transfection into testis cells and preliminary functional research of spermatogenic cell-specific gene NYD-SP12 using this method. Methods: PEI/DNA complexes were introduced into the seminiferous tubules of mouse testes using intratesticular injection. Transfection efficiency and speciality were analyzed on the third day of transfection with fluorescent microscopy and hematoxylin staining. The long-lasting expression of the GFP-NYD-SP12 fusion protein and its subcelluar localization in spermatogenic cells at different stages were analyzed with fluorescent microscopy and propidium iodide staining. Results: With the mediation of PEI, the GFP-NYD-SP12 fusion gene was efficiently transferred and expressed in the germ cells (especially in primary spermatocytes). Transfection into Sertoli cells was not observed. The subcellular localization of the GFP-NYD-SP2 fusion protein showed dynamic shifts in spermatogenic cells at different stages during spermatogenesis. Conclusion: PEI can efficiently mediate gene transfer into spermatocytes. Thus, it might be useful for the functional research of spermatogenic-cell specific genes such as the NYD-SP12 gene. In our gtudy, the NYD-SP12 protein was visualized and was involved in the formation of acrosome during spermatogenesis. Our research will continue into the detailed function of NYD-SP12 in spermatocytes. (Asian J Androl 2006 Jan; 8: 53-59)
文摘The topside floatover installation is always a great challenge and is sensitive to environmental conditions.In this study,experimental analysis on the mating operation of the floatover installation in different wave headings is presented.The continuous mating operation using the rapid transfer technique was experimentally simulated with the assistance of the jacking system and the ballast system.In the continuous transfer modeling,the topsides loads were transferred onto the jacket by several consecutive steps,including the first rapid jack-down for the 30%loads,continuous 30%−70%load transfer and the second repaid jack-down for the remaining 30%loads.Motions of the barge and the topsides as well as loads on the Deck Support Unite(DSU)and the Leg Mating Unite(LMU)in different wave headings were measured.Experimental results illustrated the complex motion behavior and load characteristics of the continuous transfer operation.Results indicate that the rapid jack-down operations will lead to impact loads and larger lateral DSU loads.The bow quartering seas are much more dangerous as it gives rise to the larger motions and loads.Comparisons with the traditional steady-state modeling indicate that the continuous transfer modeling has greater advantages over the steady-state modeling on predicting the loads.
基金Project supported by the National Key Research and Development Program of China(Grant No.2016YFA0301700)the National Natural Science Foundation of China(Grant Nos.61590932,11774333,61674132,11674300,11575172,and 11625419)+2 种基金the Anhui Provincial Initiative in Quantum Information Technologies,China(Grant Nos.AHY080000 and AHY130300)the Strategic Priority Research Program of the Chinese Academy of Sciences(Grant No.XDB24030601)the Fundamental Research Funds for the Central Universities,China
文摘Monolayer transition-metal dichalcogenides (TMDs) are considered to be fantastic building blocks for a wide variety of optical and optoelectronic devices such as sensors, photodetectors, and quantum emitters, owing to their direct band gap, transparency, and mechanical flexibility. The core element of many conventional electronic and optoelectronic devices is the p-n junction, in which the p- and n-types of the semiconductor are formed by chemical doping in different regions. Here, we report a series of optoelectronic studies on a monolayer WSe2 in-plane p-n photodetector, demonstrating a low- power dissipation by showing an ambipolar behavior with a reduced threshold voltage by a factor of two compared with the previous results on a lateral electrostatically doped WSe2 p-n junction. The fabrication of the device is based on a polycarbonates (PC) transfer technique and hence no electron-beam exposure induced damage to the monolayer WSe2 is expected. Upon optical excitation, the photodetector demonstrates a photoresponsivity of 0.12 mA.W-1 and a maximum external quantum efficiency of 0.03%. Our study provides an alternative platform for a flexible and transparent two- dimensional photodetector, from which we expect to further promote the development of next-generation optoelectronic devices.
文摘The gene encoding nonstructural protein Npro of BVDV was amplified by PCR,then was inserted into pEGFP-C1 vector.The 293 cells were transfected in vitro with recombinant plasmid by liposome.Npro-EGFP fusion protein was viewed directly with fluorensce microscope and mRNA expression of Npro was detected by reverse transcription-polymerase chain reaction(RT-PCR).Results showed that recombinant plasmid was confirmed to be constructed correctly by PCR,restriction enzyme digestion and DNA sequencing;meanwhile,the gene carried was expressed in 293 cells.The fusion protein had properties of both the two Npro and enhanced green fluorescent proteins(EGFP) and distributed in the cytoplasm.This research lays a foundation for further researches which explored Npro gene function in the process of viral replication and synthesis.
基金the National Key Research and Development Program of China(No.2019YFB2204500)the Science,Technology and Innovation Action Plan of Shanghai Municipality(No.1914220370)。
文摘Parkinson’s disease patients suffer from disorders of speech.The most frequently reported speech problems are weak,hoarse,nasal or monotonous voice,imprecise articulation,slow or fast speech,difficulty starting speech,impaired stress or rhythm,stuttering,and tremor.To improve the speech quality and assist the patient with speech rehabilitation therapy,we have proposed the speech recognition model for Parkinson’s disease patients using transfer learning technique(PSTL),where we have pre-trained the long short-term memory(LSTM)neural network model with our developed publicly available dataset that has been obtained from healthy people through the social media platform.Then,we applied the transfer learning technique to improve the performance of the PSTL framework.The frequency spectrogram masking data augmentation method has been used to alleviate the over-fitting problem so that the word error rate(WER)is further reduced.Even with a limited dataset,our proposed model has effectively reduced the WER from 58% to 44.5% on the original speech dataset and 53.1% to 43% on the denoised speech dataset,which demonstrated the feasibility of our framework.
基金financial support provided by World Bank Robert S.Mc Namara Fellowship Program 2015 to carry out this research abroad
文摘Extracts of elephant grass (Penniseturn purpureum) blended with some intensifier halides like ammonium chloride (AMC) and potassium iodide (PTI) were investigated as corrosion inhibitor for mild steel. The corrosion process was monitored in 3.5% HCI by mass loss and electrochemical techniques at 30, 40, 50, 60 and 90 ℃. Addition of AMC and PTI increased the inhibition efficiency with the highest inhibition efficiency obtained with PTI blend- ed extract. The blends behaved as mixed type inhibitors and were spontaneously adsorbed on mild steel surface in exothermic nature. Synergistic parameters of the intensifier ions revealed cooperative effect. Kinetic data treatment indicated increase in energy barrier by intensifier ions. The results demonstrate that elephant grass extract blended with halide ions can act as alternative ecofriendly inhibitor for mild steel at elevated temperatures.
文摘OBJECTIVE: To investigate the feasibility and features of 3 catheter-mediated approaches of gene transfer into heart, including direct myocardial injection (DMI), coronary artery perfusion (CAP), and intrapericardial cavity injection (ICI). METHODS: Fifteen dogs were used, and 0.3 ml (1 x 10(9) pfu) of an adenovirus (Adex1SR LacZ) was injected into the heart by 3 methods. The dogs were killed 5 days following injection, and gene expressions in heart and liver were evaluated by histochemical analysis. RESULTS: The results showed that (1) the CAP method was relatively less damaging and induced sparse LacZ expression in the myocardium, and the gene expression was also found in both vessels within the myocardium and liver; (2) gene transfer by DMI resulted in intense LacZ expression around the injection accompanied by a local inflammatory response; (3) LacZ expression elicited by ICI was detected in either the inner surface of the parietal pericardium or epicardial surface of the heart, and also in the myocardium underlying the visceral pericardium. CONCLUSION: Three catheter-mediated methods of gene transfer into the heart may be used and a reasonable approach should be chosen according to purpose.
基金ThisworkwassurpportedbyScientificCommitteeFoundationofShanghai (No 9941190 44 )
文摘OBJECTIVE: To determine the efficacy of adenovirus mediated suicide gene transduction combined with prodrug 5-fluorocytosine (5FC) as a therapeutic protocol for pancreatic cancer. METHODS: Cytosine Deaminase(CD) gene was cloned into pAdTrack-CMV-CD, pAdTrack-CMV-CD and pAdEasy-1 were recombined in bacteria. The newly recombined adenovirus (Ad)-CD containing green fluorescent protein (GFP) were packaged and propagated in 293 cells and purified by cesium chloride gradient centrifugation. Human pancreatic carcinoma cell line-Patu8988 was infected with this virus, then 5FC was added. XTT assay was used to estimate relative numbers of viable cells. In vivo model of pancreatic cancer was established by injecting 1.0 x 10(7) Patu8988 cells subcutaneously in Balb/c nude mice. When tumors were palpable, Ad-CD was injected into each tumor and 5FC was administered. RESULTS: Positive clones were selected using endonuclease to digest the recombinants and the concentration of viral liquids containing the CD gene was 2 x 10(11) pfu /ml. Significant cytotoxic activity as shown for 5FC in the CD gene transduced 8988 cell line, while little effect was found in the nontransduced pancreatic carcinoma cells. Antitumor effect was observed in Patu8988 xenograft nude mice with in situ CD gene transduction. CONCLUSIONS: CD gene mediated by adenovirus has high infectivity and may be useful for gene therapy in pancreatic carcinoma. These data demonstrate the use of an enzyme prodrug strategy in experimental pancreatic cancer.
基金agrantfromtheEducationAssociationofJiangsuProvince ,China (No .98JKB32 0 0 0 8)
文摘OBJECTIVE: To investigate the feasibility of endothelialization of bioprosthesis by transfer of vascular endothelial growth factor (VEGF) gene. METHODS: Bovine pericardium treated with glutaraldehyde and L-glutamic acid was positioned into the pig right atrium. pcD(2)/hVEGF(121) gene (1 mg) was transferred into the right ventricular myocardium using surgical sutures Reverse transcri ption polymerase chain reaction (RT PCR) was employed to evaluate the expression of myocardial VEGF mRNA. The determination of concentrations of VEGF protein in blood from both the right atrium and peripheral vein, and histological and ultrastructural analysis of implanted bovine pericardium were completed simultaneously. RESULTS: The concentration of VEGF derived from the right atrium in pcD(2)/hVEGF(121) group was significantly higher than that in the pcD(2) group 10 days after VEGF gene transfer (P
文摘OBJECTIVE: To explore the inhibitory effect of human endostatin gene mediated by retroviral vector on the growth of human liver carcinoma. METHODS: A recombinant retroviral plasmid containing human endostatin gene and signal peptide was engineered and transferred into PA317 cells to produce retrovirus. Human liver carcinoma cells (SMMC7721) were infected with the above retrovirus to build a stable endostatin-transfected liver carcinoma cell line (SMMC-endo). The control liver carcinoma cell line (SMMC-pLncx) was developed in a similar way except that the plasmid was replaced by an empty retroviral vector. Immunohistochemistry and Western blot were used to test the expression and secretion of human endostatin. The biological activity of the expressed human endostatin was assessed by endothelial cell proliferation assay. The growth rates of SMMC-endo and control SMMC-pLncx cells in vivo and in vitro were also observed. RESULTS: The expression and secretion of human endostatin by endostatin-transfected SMMC-endo cells were confirmed by immunohistochemistry and Western blot. Compared with the control group, concentrated supernatant of SMMC-endo cells remarkably inhibited the proliferation of human umbilical vein endothelial cells by 48%, significantly higher than the inhibition by the control (10.2%; P
文摘OBJECTIVE: To specifically deliver the therapeutic gene to cancer cells and construct target retroviral vectors by inserting the single-chain variable antibody fragment into the retroviral envelope. METHODS: Single-chain antibody expression vector pET -20bScfv was constructed. Binding activity of the genetically engineered single-chain variable antibody fragment was verified by enzyme-linked immunosorbent assay (ELISA) and Western blot. At the same time, by means of polymerase chain reaction (PCR)-directed mutagenesis, the appropriate cloning site EcoT22/Sal was generated at the N-terminus of receptor-binding SU domain in the MoMLV env polypeptide. Then the single- chain antibody gene, encoding a functional antibody, was inserted into the cloning site. The Scfv-env fusion gene fragment was subcloned into CMV expression vector. The Lac-Z retrovirus that co-displayed the Scfv-env chimeric protein and wild-type env protein was produced by transfection of Psi 2 cells with retroviral plasmid and the fusion gene expressing plasmid.To confirm the specificity of the recombinant retrovirus, infection assays and competitive inhibition assays were performed. RESULTS: The results of ELISA and Western blot showed that the genetically engineered single-chain variable antibody fragment could bind to the SHG44 cells surface membrane antigen. Virus-binding assay, viral infection and competitive inhibition assays confirmed that the harvested virus efficiently bound to and infected SHG44 cancer cells expressing the relative membrane antigen specially via the recognition of the target antigen. CONCLUSION: These results imply that insertion of Scfv into the retroviral envelope can specifically deliver the interested gene into specific antigen-producing cancer cells.