A region-level contrastive learning-based deep model for glomerular ultrastructure segmentation on electron microscope images / 南方医科大学学报

OBJECTIVE@#We propose a novel region- level self-supervised contrastive learning method USRegCon (ultrastructural region contrast) based on the semantic similarity of ultrastructures to improve the performance of the model for glomerular ultrastructure segmentation on electron microscope images.@*METHODS@#USRegCon used a large amount of unlabeled data for pre- training of the model in 3 steps: (1) The model encoded and decoded the ultrastructural information in the image and adaptively divided the image into multiple regions based on the semantic similarity of the ultrastructures; (2) Based on the divided regions, the first-order grayscale region representations and deep semantic region representations of each region were extracted by region pooling operation; (3) For the first-order grayscale region representations, a grayscale loss function was proposed to minimize the grayscale difference within regions and maximize the difference between regions. For deep semantic region representations, a semantic loss function was introduced to maximize the similarity of positive region pairs and the difference of negative region pairs in the representation space. These two loss functions were jointly used for pre-training of the model.@*RESULTS@#In the segmentation task for 3 ultrastructures of the glomerular filtration barrier based on the private dataset GlomEM, USRegCon achieved promising segmentation results for basement membrane, endothelial cells, and podocytes, with Dice coefficients of (85.69 ± 0.13)%, (74.59 ± 0.13)%, and (78.57 ± 0.16)%, respectively, demonstrating a good performance of the model superior to many existing image-level, pixel-level, and region-level self-supervised contrastive learning methods and close to the fully- supervised pre-training method based on the large- scale labeled dataset ImageNet.@*CONCLUSION@#USRegCon facilitates the model to learn beneficial region representations from large amounts of unlabeled data to overcome the scarcity of labeled data and improves the deep model performance for glomerular ultrastructure recognition and boundary segmentation.

Palm vein recognition based on end-to-end convolutional neural network / 南方医科大学学报

We propose a novel palm-vein recognition model based on the end-to-end convolutional neural network. In this model, the convolutional layer and the pooling layer were alternately connected to extract the image features, and the categorical attribute was estimated simultaneously via the neural network classifier. The classification error was minimized via the mini-batch stochastic gradient descent algorithm with momentum to optimize the feature descriptor along with the direction of the gradient descent. Four strategies including data augmentation, batch normalization, dropout, and L2 parameter regularization were applied in the model to reduce the generalization error. The experimental results showed that for classifying 500 subjects form PolyU database and a self-established database, this model achieved identification rates of 99.90% and 98.05%, respectively, with an identification time for a single sample less than 9 ms. The proposed approach, as compared with the traditional method, could improve the accuracy of palm vein recognition in clincal applications and provides a new approach to palm vein recognition.

Application of N-methyl-aspartate receptor antibody level, brainstem auditory evoked potential and magnetic resonance imaging in the differential diagnosis of encephalitis in children / 中华实验和临床病毒学杂志

Objective@#To explore the clinical value of serum N-methyl-aspartate receptor (NMDAR) antibody level, brainstem auditory evoked potential (BAEP) and magnetic resonance imaging (MRI) in the differential diagnosis of viral encephalitis and anti-NMDAR encephalitis.@*Methods@#The clinical data of 68 children patients with encephalitis were retrospectively analyzed. The patients diagnosed with viral encephalitis were included in V group (n=52), and the patients diagnosed with anti-NMDAR encephalitis were included in N group (n=16). The clinical characteristics, serum NMDAR antibody level, and BAEP and MRI findings were compared between the two groups.@*Results@#The age, disease duration, abnormal behavior rate, sleep disorder rate and epileptic seizure rate in V group were significantly lower than those in N group [(6.62±1.20)Y/O vs.(8.46±1.85)Y/O, (3.53±0.71)d vs.(4.49±0.82)d, 30.77%(16/52)vs. 75.00%(12/16), 21.15%(11/52)vs. 62.50%(10/16), 26.92%(14/52)vs. 56.25%(9/16), t=4.681, t=4.560, χ2=9.882, χ2=7.958, χ2=4.701], while the abnormal rate of video EEG was significantly higher than that in N group [51.92(27/52)vs. 81.25%(13/16), χ2=4.345] (all P<0.05). There were no significant differences in gender, rates of prodromic infection symptoms, cognitive impairment, fever, headache, convulsion and incidence rate of meningeal irritation sign (P>0.05). The serum NMDAR antibody level in V group was significantly lower than that in N group [(3.40±0.69) ng/ml vs.(13.95±2.78) ng/ml t=25.319)] (P<0.05). There were no significant differences in the BAEP apparent involvement range and central auditory neurological damage between the two groups (P>0.05), but the peripheral auditory nerve damage and total BAEP abnormality rate in V group were significantly lower than those in N group [3.85%(4/104)vs. 21.88%(7/32), 6.73%(7/104)vs. 28.12%(9/32), 30.77%(16/52)vs. 62.50%(10/16), χ2=10.699, χ2=10.790, χ2=5.216] (all P<0.05). There were no significant differences in MRI signal intensity, lesion involvement range and total abnormal rate between the two groups (all P>0.05).@*Conclusions@#There were significant differences in serum NMDAR antibody level and BAEP test results among children patients with viral encephalitis or anti-NMDAR encephalitis, and they are helpful for early differential diagnosis.

Epidemiological analysis of complications in preterm infants / 中国基层医药

Objective To explore the epidemiological analysis of complications in premature infants.Methods From January 2017 to March 2018,1800 premature babies in Ningbo Women and Children Hospital were selected in the study.The clinical data of pregnant women,premature infants and premature complications,and so on were investigated,and summarized epidemiology of premature infant complications.Results The probability of premature birth was 9.09％.The differences between different gestational age(x2 =2 481.34) and different body weight (x2 =3 088.21) were statistically significant (all P ＜ 0.05).Premature rupture of membranes occurred as the main one of the common factors lead to premature birth,the fetal distress was also more common factors,so in different gestational age,the difference was not statistically significant (P ＞ 0.05).However,compared with other factors,the difference of premature infants at different gestational weeks was statistically significant (P ＜ 0.05).Conclusion Epidemiological study of preterm infants,can promote their perinatal management level and quality,and then pointed to strengthen perinatal health education and health care,for the effective prevention and treatment of common diseases of perinatal,key management and monitoring work earnestly strengthen the high-risk pregnancy,attaches great importance to the establishment of collaborative relationship between made in pediatric claims intrauterine transhipment,making pregnant women can be produced in the hospital for treatment for premature babies,and reduce the mortality and morbidity rates of premature and low birth weight,can reduce the risk of intellectual disability,eventually making the birth population overall quality improved.

Preliminary study on the short－term and long－term effects of the inhibition of hypoxia inducible factor－1 alpha decomposition and activation of Wnt signaling pathway in brain injury in neonatal rats / 中国新生儿科杂志

Objective To investigate the protective effect of inhibition of hypoxia inducible factor－1 alpha (HIF－1α) and activation of Wnt signaling pathway on brain injury in neonatal rats.Method A total of 312 newborn SD rats were used to establish cerebral white matter damage model (WMD) on day 3.And they were randomly assigned into the model group (WMD group), post－modeling HIF－1αdecomposition inhibition group (PHI group), and post－modeling HIF－1αdecomposition inhibition with activation of Wnt signaling pathway group (PHI+activated Wnt group ) and post－modeling HIF－1αdecomposition inhibition with inhibition of the Wnt signaling pathway group (PHI+inhibition Wnt group).Brain tissues were taken on day 1, 3, 7 and 14 after modeling, respectively.The changes of oligodendrocyte precursor cell (OPC) and oligodendrocyte ( OL ) in brain tissues at different time points were observed by tissue immunofluorescence.The expression of HIF－1αprotein in the brain tissue of each time point was measured by western blot technique.The mRNA level of HIF－1αand Wnt7a in the brain tissue of each time point was detected by RT－qPCR technique.Behaviors of rats were tested by the suspension experiment , the open field experiment and the dark experiment , at 28 d after modeling.Result The numbers of OPC on 1 d and 3 d after modeling and the number of OL on 7 d and 14 d after modeling in PHI +activated Wnt group were significantly higher than the other three groups.The content of HIF－1αin WMD group was the least on 1, 3, 7 and 14 d, but the content of PHI+activated Wnt group was the highest , and the difference was statistically significant (P＜0.05).On 1, 3, 7 and 14 d after modeling, the expression level of HIF－1αand Wnt7a in PHI+activated Wnt group were higher than those in the other three groups , the difference was statistically significant (P＜0.05), and Wnt7a expression was positively correlated with the change of HIF－1α(P＜0.05).There was no significant difference of suspension experiment at 28 d after modeling between groups. Compared with other groups , WMD group had the lowest score on open field experiment ( P＜0.05).The PHI+activation Wnt group had better memory function , and then the PHI group , WMD group.The latent time of dark experiment were significantly shorter in PHI +acitivation Wnt group.There were more mistaken time of dark experiment in WMD group compared with PHI +activation Wnt group.Conclusion Inhibition of HIF－1αdecomposition and activation of Wnt signaling pathway have partially repair effect on brain injury in neonatal rats.

Influences of different microbial environment on gut microbiota colonization in early life of mice / 中华围产医学杂志

Objective To investigate the influences of exposure to different environmental microbes on early-life gut microbiota colonization in mice.Methods Male (n=8) and female (n=16) adult specific pathogen free (SPF) BALB/c mice were caged together at a ratio of 2:l.After conception,the mice were divided into four groups according to the environments where the offsprings were reared at three different periods (fetal period,breastfeeding period and childhood).Group A:Offsprings were kept in a SPF environment throughout the study;group B:SPF environment during fetal and breastfeeding periods,and then ordinary environment during childhood;group C:SPF environment during fetal period,and then ordinary environment during breastfeeding period and childhood;group D:ordinary environment all the time.Fecal samples were collected at the end of week 3 and 5.Total bacterial DNA was extracted from each sample and analyzed by high throughput analysis.Kruskal-Wallis and Dunn-Bonferroni test were applied for statistical anaysis.Results 1.At the end of three weeks:(1) Diversity:① Phylum level:There were significant differences in the abundance of Firmicutes,Verrucomicrobia,Proteobacteria and Actinobacteria among the four group (all P＜0.01).Compared with group C and D,group A and B showed significantly decreased abundance of Firmicutes [30.876(23.448-41.218)× 10-2,3.317(1.116-4.641) 10-2 vs 71.936(53.587-86.713)× 10-2,79.105(56.305-82.736)× 10-2],but increased abundance of Verrucomicrobia and Proteobacteria [Verrucomicrobia:17.249(9.748-35.106)× 10-2,58.883(0.017-6.047)× 10-2 vs 0.152(0.066-1.890)× 10-2,0.003(0.000-0.016)× 10-2;Proteobacteria:12.640(0.336-15.070)× 10-2,3.653(3.362-4.5955)× 10-2 vs 0.219(0.134-0.325)× 10-2,0.124(0.116-0.165) × 10-2,all P＜0.05 or 0.01].② Genus level:There were significant differences in the abundance of Lactobacillus,Akkermansia and Bacteroides among the four groups (all P＜0.01).Compared with group C and D,group A and B showed significantly decreased abundance of Lactobacillus [19.283(8.618-31.541)× 10-2,0.339(0.264-22.278) × 10-2 vs 58.414(34.874-71.942)× 10-2,66.007(55.141-76.940)× 10-2],but increased abundance of Akkermansia,Bacteroides and Klebsiella [Akkermansia:17.247(9.748-35.106)× 10-2,58.883(0.017-60.475)× 10-2 vs 0.152(0.066-1.890)× 10-2,0.003(0.000-0.017)× 10-2;Bacteroides:3.978(0.683-25.171)× 10-2,8.216(6.023-9.946)× 10-2 vs 0.141(0.061-0.281)× 10-2,0.568(0.149-1.455)× 10-2;Klebsiella:0.209(0.050-8.888)× 10-2,1.402(0.865-1.692)× 10-2 vs 0.003(0.000-0.039) 10-2,0.000(0.000 0.001)× 10-2,all P＜0.05 or 0.01].(2) Alpha diversity:Significant differences were found in operational taxonomic unit (OTU) and Chaol index (P＜0.05),but not in Shannon index among the four groups (P＞0.05).The OTUs of group A and B were significantly lower than that of group D [246(221-348),257(209-280) vs 387(324-478),P=0.045 and 0.008,respectively].2.At the end of five weeks:(1) Diversity:① Phylum level:There were significant differences in the abundance of Firmicutes,Verrucomicrobia and Proteobacteria among the four groups (P＜0.05 or 0.01).The abundance of Firmicutes in gut microbiota in group A was lower than that in group B,C and D [13.765(64.181-24.238)× 10-2 vs 48.912(37.280-59.466)× 10-2,86.065(50.149-89.856) × 10-2,53.847(31.946-72.936) × 10-2],while that of Verrucomicrobia was higher [58.089(22.459-61.285)× 10-2 vs 0.001(0.000-0.005)× 10-2,0.000(0.000-0.001)× 10-2,0.003(0.000-0.006)× 10-2],all P＜0.05 or 0.01.② Genus level:There were significant differences in the abundance of Lactobacillus and Akkermansia among the four groups (P＜0.01).The abundance of Lactobacillus in gut microbiota in group A was lower than that in group B,C and D[1.755(0.805-8.833)× 10-2 vs 26.391(17.550-37.265)× 10-2,70.688(45.713-77.953) × 10-2,28.675 (15.660-57.224) × 10-2],while that of Akkermansia was higher [58.089(22.460-61.285)× 10-2 vs 0.000(0.000-0.006)× 10-2,0.000(0.000-0.001)× 10-2,0.003(0.000-0.006)× 10-2,all P＜0.05 or 0.01].(2) Alpha diversity:There were significant differences in OTU,Chaol and Shannon index among the four groups (P＜0.05 or 0.01).The OTU of group A was lower than that of group B,C and D [268(241-410) vs 438(380-516),562(533-588),546(473-599)],and the OTU,Chaol and Shannon index of group B were all lower than those of group C and D [OTU:438(380-516) vs 562(533-588),546(473-599);Chaol index:1 033(883-1 181) vs 1 285(1 220-1 338),1 328(1 155-1 516);Shannon index:3.85(3.25-4.50) vs 4.28(3.30-5.11),4.17(3.62-4.38),all P＜0.05 or 0.01].Conclusions Early-life exposure to different environments has an obvious impact on the diversity and composition of intestinal microbiota in mice.The less clean the living environment is,the more diverse the gut microflora will be.Furthermore,the window of opportunity for gut microbiota colonization seems to be related to breastfeeding period.

Association of serum levels of matrix metalloproteinases and their tissue inhibitors with mortality in patients with malignant middle cerebral artery infarction / 中国医师杂志

Objective To investigate the relationship between serum levels of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMP) and the mortality in patients with malignant middle cerebral artery infarction.Methods A total of 74 patients with malignant middle cerebral artery infarction (MMCAI) was assigned to research group,74 healthy volunteers were recruited to serve as healthy controls,and 242 patients with other cerebral infarction were recruited to serve as case controls.Quality-of-life questionnaire and clinical examination were used to collect the information of participants.Endpoint was 30-day mortality.Results The research group showed higher serum levels of MMP-9,MMP-10,and TIMP-1 relative to the case control group,and the case control group showed higher serum levels of MMP-9,MMP-10,and TIMP-1 relative to the healthy control group (P ＜ 0.05).Compared to survivor ones,non-surviving MMCAI patients showed lower Glasgow Coma Scale score (P ＜ 0.05),higher serum levels of tumor necrosis factorα (TNF-α),MMP-9,MMP-10,and TIMP-1 (P ＜ 0.05).Multiple logistic regression analysis showed that serum levels of TNF-α,MMP-9,MMP-10,and TIMP-1 were associated with 30-day mortality,and the OR were 1.09,1.18,1.13,and 1.32,respectively.Conclusions Serum levels of MMP-9,MMP-10,and TIMP-1 levels in MMCAI patients were associated with mortality,and could be used as an auxiliary index of mortality in MMCAI patients.

Edge-detecting operator-based selection of Huber regularization threshold for low-dose computed tomography imaging / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare two methods for threshold selection in Huber regularization for low-dose computed tomography imaging.</p><p><b>METHODS</b>Huber regularization-based iterative reconstruction (IR) approach was adopted for low-dose CT image reconstruction and the threshold of Huber regularization was selected based on global versus local edge-detecting operators.</p><p><b>RESULTS</b>The experimental results on the simulation data demonstrated that both of the two threshold selection methods in Huber regularization could yield remarkable gains in terms of noise suppression and artifact removal.</p><p><b>CONCLUSION</b>Both of the two methods for threshold selection in Huber regularization can yield high-quality images in low-dose CT image iterative reconstruction.</p>

HLA-DPB1 gene polymorphism and susceptibility to tuberculosis in Xinjiang Shihezi Han population:a case-control study / 中国免疫学杂志

Objective:In this study,we examined the HLA-DPB1 alleles in patients with tuberculosis,and health individuals attempt to investigate the association between the polymorphism of HLA-DPB1 gene and pulmonary tuberculosis in Han population in Shihezi area in Xinjiang uygur autonomous region of China.Methods:High-resolution typing of DPB1 was performed by the sequence-based typing ( SBT) method using the SBT-HLA-DPB1 generic DNA typing kit.Results:In the controls ,17 HLA-DPB1 alleles were ob-served,the HLA DPB1*0501 (28.1%) and HLA DPB1*0201(27.6%) frequency was significantly higher than other sites ,the first and second respectively.The frequency of HLA-DPB1* 0201 was observed significantly increased in patient group compared with control group ( P<0.05 );the frequency of HLA-DPB1* 0501 was observed significantly decreased in patient group compared with control group ( P<0.05 ).Conclusion: The gene frequencies of HLA-DPB1 in Xinjiang Shihezi Han population are roughly in consistence with other northern Chinese Han population;the HLA-DPB1*0201 may be the protective factor to pulmonary tuberculosis , and HLA-DPB1*0501 may be susceptible to pulmonary tuberculosis in Han population from the Xinjiang uygur autonomous region of China.

Relationship of gallbladder emptying function and its hemodynamics in diabetic patients / 中国糖尿病杂志

Objective To explore the relationship of gallbladder emptying function and its hemodynamics in diabetic patients. Methods Gallbladder volume ,emptying function and its arterial blood flow were measured by ultrasound. Results The gallbladder volumes of fasting (FV ) and postprandial (RV) increased and the ejection fraction (EF) of gallbladder was decreased in DM group than in NC group (P<0.01). In DM group ,the gallbladder artery’s resistance index (RI) was higher than in NC group [(0.61 ± 0.06) vs (0.70 ± 0.08)](P<0.01). Minimum diastolic blood flow velocity (Vmin) was lower than in NC group [(8.72 ± 1.43) vs (6.05 ± 1.89)cm/s](P<0.01). RI was associated with HbA1c (r= 0.827 ,P< 0.01) and LDL‐C(r= 0.820 ,P< 0.01). Conclusion Gallbladder emptying dysfunction in diabetic patients is associated with the hemodynamics of gallbladder.

Magnetic field numerical calculation and analysis for magnetic coupling of centrifugal blood pump for extracorporeal circulation / 生物医学工程学杂志

This paper mainly studies the driving system of centrifugal blood pump for extracorporeal circulation, with the core being disc magnetic coupling. Structure parameters of disc magnetic coupling are related to the ability of transferring magnetic torque. Therefore, it is necessary to carry out disc magnetic coupling permanent magnet pole number (n), air gap length (L(g)), permanent magnet thickness (L(m)), permanent magnet body inside diameter (R(i)) and outside diameter (R(o)), etc. thoroughly. This paper adopts the three-dimensional static magnetic field edge element method of Ansys for numerical calculation, and analyses the relations of magnetic coupling each parameter to transmission magnetic torque. It provides a good theory basis and calculation method for further optimization of the disc magnetic coupling.

Tsallis entropy-based prior for PET reconstruction / 生物医学工程学杂志

Maximum a Posteriori (MAP) method has been widely applied to the ill-posed problem of image reconstruction. The choice of prior is the crucial point on MAP methods. However, the most conventional priors will lead to a blurring of the whole image or cause ladder-like artifacts. We therefore proposed a Tsallis entropy-based prior for positron emission tomography (PET) iterative reconstruction in MAP framework. The method uses a Tsallis entropy-based prior to eliminate the uncertainty between prior information and the estimated images. We tested this method in the phantom image, compared it with the traditional prior methods. the results showed that the proposed algorithm could suppress noise and obtain better reconstructed image quality.

Study on optimal selection of structure of vaneless centrifugal blood pump with constraints on blood perfusion and on blood damage indexes / 生物医学工程学杂志

This paper is aimed to study the optimal selection of structure of vaneless centrifugal blood pump. The optimal objective is determined according to requirements of clinical use. Possible schemes are generally worked out based on structural feature of vaneless centrifugal blood pump. The optimal structure is selected from possible schemes with constraints on blood perfusion and blood damage indexes. Using an optimal selection method one can find the optimum structure scheme from possible schemes effectively. The results of numerical simulation of optimal blood pump showed that the method of constraints of blood perfusion and blood damage is competent for the requirements of selection of the optimal blood pumps.

Prokaryotic expression and antigen characteristics of EB virus latent membrane protein 2 ( EBV-LMP2) multi-epitopes / 中华微生物学和免疫学杂志

Objective To express in prokaryotic system and to analyze the antigenic specificity of EB virus(EBV) latent membrane protein 2(LMP2) multi-epitopes gene rich of T cell and B cell epitopes.Methods Using on-line prediction service, T cell epitopes and B cell epitopes of EB virus latent membrane protein 2 were predicted. The genes rich of CTL and th cell epitopes were selected as the candidate gene sequences, while B cell epitopes around them were taken into account. The finial selected multi-epitope gene was synthesized after being optimized according to prokaryotic codon bias and inserted into prokaryotic expression vector pET32a( + ) to get the recombinant plasmid: pET32a( + )/EBV-LMP2 multi-epitopes. After transformed into E. coli BL21 (DE3) and induced by IPTG, the target multi-epitopes gene can be expressed as Trx-His fusion protein. The expression products can be identified by SDS-PAGE and Western blot. Moreover, rabbit serum antibody to EBV membrane protein and nasopharyngeal carcinoma(NPC) patient serum were used respectively to detect the antigenic specificity of the multi-epitopes. Meanwhile, 6-8 weeks female BALB/c mice were immunized with EBV-LMP2 multi-epitope at 2 week intervals, three times in all, Trx-His protein and PBS were set as the control groups. At the second week after the last immunization, the mice were sacrificed. LDH and indirect ELISA were taken to detect the specific spleen CTL activityand specific IgG in serum, which reflected the immunogenicity of the EBV-LMP2 multi-epitope. Results Two amino acid sequences which locate at the LMP2 (aa195 -232 ) and LMP2 (aa419-436 ) were selected and connected in series to be the target gene. The recombinant plasmid containing EBV-LMP2 multi-epitope gene successfully constructed and the target protein was expressed in E. coli BL21 ( DE3 ). The relative molecular mass(Mr) of The expression products is about 27 × 103 , which matches up to the expected Mr. The antigenic specificity of the multi-epitopes protein was identified by Western blot and the multi-epitopes protein also can be detected by rabbit serum antibody to EBV membrane protein and NPC patient serum respectively. In the result of the animal experiment, EBV-LMP2 multi-epitope was able to induce the specific CTL activity in BALB/c mice. With the increasing of the effector: target ( E: T) 1: 5,1: 10, 1: 25, the CTL activity was also increased wih( 12.52% + 2.59% ), (21.80% + 1.08% ), (23.68% + 3.74% ) respectively; EBV-LMP2 multi-epitope was able to induce LMP2-specific antibody response(A490 =0.258 +0.040) as compared with Trx-His protein(A490 =0.095 +0.011) and PBS(A490 =0.068 +0.014,P＜0.05＝. Conclusion The EBV-LMP2 multi-epitopes gene was designed successfully and expressed precisely in prokaryotic expression system with good antigenicity and immunogenicity.

The cellular immune response produced in BALB/c mice immunized with HPV6b L1/Ct MOMP multi-epitope chimeric DNA / 中华微生物学和免疫学杂志

Objective To study on the specific cellular immune response produced in BALB/c mice immunized with human papillomavirus (HPV) type 6b capsid protein L1 and Chlamydia trachomatis (Ct) major outer membrane protein(MOMP) multi-epitope chimeric DNA (HPV6b L1/Ct MOMP multiepitope) , and the enhancement of the specific cellular immune response to Ct MOMP multi-epitope by HPV6b L1. Methods The Ct MOMP multi-epitope gene was connected to the C terminal of HPV6b L1,the gene of HPV6b L1 had been optimized according to the codon usage of eukaryotic system, and then the HPV6b L1/Ct MOMP multi-epitope chimeric gene was cloned to pcDNA3.1 ( + ) vector. After identification by restriction enzyme digestion and DNA sequencing, the recombinant plasmid was transfected into COS-7 cells, Indirect Immunofluorescence (IIF) was used to confirm the expression of proteins. Then, BALB/c mice were randomly assigned to receive (intramuscular injection) either pcDNA3.1 ( + )/HPV6b L1/Ct MOMP or pcDNA3.1 ( + )/Ct MOMP or pcDNA3.1 ( + ) or PBS ( n = 12, 150 μg/time), and the same immunization schedule was repeated third times at 2 week intervals. The level of cytokine( IFN-γ, IL-4, IL-10) -producing CD3+ T cells in spleen, the cytotoxicity of Ct MOMP-specific and HPV6b L1-specific cytotoxic T lymphocyte (CTL) in spleen were detected by intracellular cytokine staining-fluorescence activated cell sorter (ICS-FACS) and LDH release assays, respectively. Results After immunization, when the efCTL (44.56%±4.02%, 35.35% ±2.89% ) and HPV6b L1 specific cytotoxicity of CTL (27.08% ±2.04%, 21.68% ±4.06% ) in pcDNA3.1 ( + )/HPV6b L1/Ct MOMP multi-epitope chimeric DNA immunized mice, were significantly higher than that in pcDNA3.1 ( + )/Ct MOMP multi-epitope DNA (35.50%±2.68%, 30.24% ±1.75%; 12.27% ±3.36%, 9.32% ±3.07%) and other control groups(F=72.87, F=114.55, P＜0.05; F=30.04, F=10.47, P＜0.05), and Ct MOMP multi-epitope specific cytotoxicity of CTL in pcDNA3.1 ( + )/Ct MOMP multi-epitope DNA immunized mice were significantly higher than that in control groups( F = 58.85, F = 120.21; P<0.05). The level of intracellular cytokine IFN-γ in pcDNA3.1 ( + )/HPV6b L1/Ct MOMP multi-epitope DNA immunized mice(4.34% ±0.06%)was higher significantly than that in pcDNA3.1 ( + )/Ct MOMP multi-epitope DNA immunized mice(3.14% ± 0.18%, P<0.05 ) and other control groups ( F = 473.83, P<0.05 ), while, the levels of IL-4 ( F =0.97, P ＞ 0.05 ) and IL-10 ( F = 2.25, P ＞ 0.05 ) had no significant difference between groups. Conclusion Both Ct MOMP and HPV6b L1 protein specific cellular immune response could be induced in BALB/c mice immunized with HPV6b L1/Ct MOMP multi-epitope chimeric plasmid, and the HPV6b L1 gene optimized by eukaryotic codon could significantly enhance the cellular immune response induced by Ct MOMP multi-epitope gene in BALB/c mice.

Optimal Acupuncture Therapeutic Projects for Cerebral Infarction-related Motor Dysfunction of Lower Limbs / 针灸推拿医学（英文版）

Objective: to select the optimal acupuncture therapeutic projects on cerebral infarction-related motor dysfunction of lower limbs. Methods: to optimize the combination projects on 4 factors and 3 levels affecting the acupuncture effect on cerebral infarction by using orthogonal design targeting on patients with cerebral infarction-related motor dysfunction of lower limbs, and Fugl-Meyer score of limb motor function was taken as indexes. Results: The relatively optimal Fugl-Meyer score of lower limb function can be obtained within 3-day duration of cerebral infarction. Conclusions: As far as the considered factors and levels are concerned, the previously mentioned project is the optimal acupuncture therapeutic project for cerebral infarction-related motor dysfunction of lower limbs.