Objective:To analyze the association between gene single nucleotide polymorphisms ( SNP) of methylenetetrahydrofo-late reductase ( MTHFR) gene and lower extremity atherosclerotic disease ( LEAD) . Methods:The clinical data and peripheral blood were collected from 384 participants (224 LEAD cases and 160 normal controls) from Han population of Minnan Fujian. LEAD was detected with ankle brachial index ( ABI) , toe brachial index ( TBI) , color Doppler ultrasonic examination and the other imaging stud-ies. The SNP genotypes including rs1801133, rs1801131, rs2274976, rs4846048, rs3737966, rs1537515, rs4846049, rs3834044, rs13306561 and rs3737964 in the MTHFR gene were detected by matrix-assisted laser desorption ionization-time of flight ( MALDI-TOF) . Results:The genotype distributions of the ten loci were in accordance with Hardy-Weinberg equilibrium. There were 37 obvi-ous linkage disequilibrium, including the association between rs4846048 and rs3737966 (D′>0. 9) and so on. There were significant differences (P=0. 02) in GCCTCGGAAT haplotypes of MTHFR gene groups between LEAD cases and the normal groups. The results from chi-square test of allele frequencies suggested rs1801131 (OR=1. 287),rs4846048 (OR=1. 844,P=0. 02), rs3737966(OR=1. 339),rs4846049 (OR=1. 314) and rs3737964 (OR=1. 522). Significant differences (P<0. 05) were observed between LEAD and the normal groups in Cochran- Armitage trend test and Dominant gene action test of rs4846048. Conclusion: The SNP of rs1801131,rs4846048,rs3737966,rs4846049 and rs3737964 might be associated with the susceptibility of LEAD,and rs4846048 gene mutation might serve as a risk factor for LEAD in the community-based population.

Objective To investigate the effects of IL-12 coexpression level on antigen expression and immune responses induced by HBsAg DNA vaccination. Methods DNA vaccine plasmid pHBV carrying codon-optimized preS2-S gene of reference sequence CHN-HBV07-C in China was constructed. Three DNA vaccine plasmids pHBV-12i, pHBV-12l and pHBV-12h were also constructed by subcloning three different IL-12 expression cassettes with various expression strengths to plasmid pHBV, respectively. Expression levels of IL-12 and HBsAg in vaccine plasmid-tranfected 293T cells were measured by quantitative ELISA. DNA vaccines were administered intramuscularly to BALB/c mice and HBsAg-specific cellular immune responses were determined by IFN-γ ELISPOT. HBsAg-specific antibodies were tested by Chemiluminescence Quantitative Immunoassay. Results The HBsAg expression level in 293T cells was 70 ng/ml when transfected by plasmid pHBV without IL-12 expression cassette, and the HBsAg level was 18 ng/ml when transfected by plasmid pHBV-12l carrying low-level IL-12 expression cassette, whereas the HBsAg level was only 6 ng/ml when transfected by plasmid pHBV-12h carrying high-level IL-12 expression cassette.Results of DNA vaccination revealed that HBsAg-specific humoral and cellular immune responses were significantly decreased in mice administering vaccine pHBV-12h carrying high-level IL-12 expression cassette. Although HBsAg-specific antibody responses in mice inoculated with pHBV-12l were also decreased when compared with those in pHBV-vaccinated mice without IL-12 expression, the HBsAg-specific cellular immune responses were significantly increased. Conclusion High-level coexpression of IL-12 may suppress the expression of HBsAg, Whereas modest coexpression of IL-12 significantly enhanced the HBsAg-specific T cell responses induced by DNA vaccination. Therefore, it is so important to balance the expression between adjuvant and antigen to enhance the immune response.

Objective To explore the value of mammogram texture analysis in prediction of metastasis of axillary lymph nodes for breast carcinoma.Methods Mammograms and clinical data of 171 patients with breast carcinoma confirmed by pathology were retrospectively analyzed,and all patients underwent axillary lymph node dissection (ALND).Then the patients were divided into axillary lymph node metastasis group and non-metastasis group according to the result of ALND.The texture features of these lesions were statistically analyzed,including gray-level histogram texture parameters (mean value,standard deviation,skewness,kurtosis and variance) and gray-level co-occurrence matrix texture parameters (energy,entropy,correlation,inertia,inverse difference moment and contrast).Results In all of 171 breast cancer patients,96patients had axillary lymph node metastasis,while 75 patients had no metastasis.Mammograms showed negative axillary lymph nodes in 119 patients and positive axillary lymph nodes in 52 patients,and the sensitivity and specificity of mammograms in the diagnosis of positive axillary lymph nodes was 48.96％ (47/96) and 93.33％ (70/75),respectively.Mammogram texture analysis showed the values of energy,entropy,inverse difference moment and correlation in axillary lymph node metastasis group were higher than those in non-metastasis group,while the values of inertia and contrast in the axillary lymph node metastasis group were lower than those in non-metastasis group (all P＜0.05).The rest texture parameters had no significant differences between two groups (all P＞0.05).Area under curve (AUC) for texture parameters of energy,entropy,inertia,inverse difference moment,correlation and contrast was 0.610,0.610,0.374,0.599,0.612 and 0.421 (all P＜0.05),respectively.AUC of mammography,mammogram texture features,and the combination of mammography and texture features was 0.711,0.676 and 0.787 (all P＜0.05),respectively.The sensitivity and specificity of mammogram texture features,the combination of mammography and texture features in diagnosis of axillary lymph nodes metastasis was 62.5％ and 64.6％,66.7％ and 82.7％,respectively.Conclusion Mammogram texture parameters are helpful for predicting axillary lymph node metastasis,and the combination of mammography and texture features can improve diagnostic efficiency of axillary lymph node metastasis.

Objective:To analyze the epidemiological and clinical characteristics of children with brucellosis, and to provide a practical basis and experience for clinical diagnosis and treatment of brucellosis.Methods:Retrospective analysis was used to collect clinical data of children with brucellosis diagnosed at the Children's Hospital Affiliated to Zhengzhou University from May 2015 to October 2017, and their epidemiological characteristics, clinical manifestations, laboratory tests, and clinical diagnosis and treatment were summarized.Results:A total of 24 children were included, including 14 males and 10 females, with an average age of 6 years (1 year 2 months to 12 years old). Except February, onset throughout the year, higher incidence was from May to July (14 cases, 58.33%). The exposure history of the children was mainly exposure to cattle and sheep and consumption of beef and mutton (18 cases, 75.00%). The main clinical manifestations were fever in 24 cases (100.00%), bone and joint pain in 14 cases (58.33%), hepatomegaly in 9 cases (37.50%), splenomegaly in 7 cases (29.17%). Tube agglutination test (SAT) was positive in 21 cases (87.50%), weakly positive in 1 case (4.17%) and negative in 2 cases (8.33%). Brucella was detected in all 24 cases by microbial culture, and in 18 cases (75.00%) by blood culture. Eighteen cases (75.00%) had liver dysfunction. Thirteen cases were misdiagnosed, and the misdiagnosis rate was as high as 54.17%. Twenty-two cases had been cured after treatment, 2 cases relapsed and recovered after continued treatment. Conclusions:Children with brucellosis have diverse epidemiology and clinical features, and are easily misdiagnosed. For children with fever, bone and joint pain and exposure history, pediatricians should be alert to the possibility of brucellosis, conduct microbiological and serological tests, in order to timely, accurate and standardized diagnosis and treatment of children with brucellosis.

Objective: Sex differences have been observed in many aspects of schizophrenia, including cognitive deficits. Despite extensive research into the relationship between metabolic factors and cognitive deficits in schizophrenia, few studies have explored the potential sex difference in their association. Methods: We recruited 358 schizophrenia patients and 231 healthy controls. The participants underwent measurements of body mass index (BMI), waist circumference, blood pressure, triglycerides, high-density lipoprotein cholesterol, and fasting blood glucose. Metabolic risk factors included abdominal obesity, hypertension, hyperglycemia, and dyslipidemia. A collection of these metabolic risk factors has been defined as metabolic syndrome. These diagnoses were based on the criteria of the National Cholesterol Education Program’s Adult Treatment Panel III. Cognitive performance was measured using the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS). A descriptive analysis, difference analysis, and linear regression model were used to identify the metabolic risk factors for cognitive function in schizophrenia. Results: Our findings revealed sex differences in the rate of abdominal obesity and hypertension in schizophrenic patients. Additionally, we observed sex differences in the association between metabolic risk factors and cognitive impairment in schizophrenia. Specifically, hyperglycemia was associated with the immediate memory index score of RBANS in male patients, while dyslipidemia was associated with language, attention, delayed memory index scores, and RBANS total score in female patients. Conclusion Our results suggest that sex should be considered when evaluating the impact of metabolic disorders on the cognitive function of schizophrenic patients. Moreover, our study identifies hyperglycemia and dyslipidemia as potential targets for precise treatment by sex stratification, which could benefit the improvement of cognitive impairment in schizophrenic patients.

To prepare recombinant human retinol binding protein 4 (RBP4) by using the baculovirus expression system and to detect its immunogenicity, the fusion DNA fragment of secretory signal peptide SS64 and human RBP4 gene was subcloned into a baculovirus transfer vector pFastBac-dual(pFBd), and the corresponding recombinant transfer plasmid was transformed into E. coli strain DH10bac, after transposition recombinant shuttle bacmid was screened out. The logarithmic phase Sf9 cells were transfected with the recombinant bacmid and then the recombinant baculovirus containing hRBP4 expression box were generated. After amplification of recombinant baculovirus, the recombinant baculovirus seeds were obtained. To express human RBP4, logarithmic phase Sf9 cells were infected with the virus seeds and SDS-PAGE and Western blotting were used to detect and identify the expression. Finally, to prepare a batch of RBP4 protein, logarithmic phase Sf9 cells in suspension culture were infected with recombinant baculovirus seeds and the supernatant was harvested after 120 hours post-infection for purification. Finally for preparation of polyclonal antibody and evaluation of immunogenicity, the recombinant hRBP4 from insect cells and from E. coli were immunized rabbits. Restriction enzyme digestion and sequencing confirmed that the recombinant baculovirus transfer plasmid was constructed correctly, and subsequently recombinant RBP4-bacmid was generated successfully. SDS-PAGE and Western blotting analysis suggested that human RBP4 protein was highly expressed in Sf9 cells with the molecular weight of approximately 23 kDa. The recombinant RBP4 protein could be secreted into the medium efficiently, and the expression level was calculated amount of 100 mg/L. Finally the rabbit antiserum was harvested after recombinant RBP4 immunization, therein the titer of antiserum against baculovirus recombinant RBP4 is 1:100 000 whereas the titer of antiserum against E. coli recombinant RBP4 is only 1:10 000. Overall, human RBP4 was high efficiently expressed successfully with good antigenicity in baculovirus system, and high affinity antiserum was obtained. A solid foundation was laid for the next step of the preparation of human serum RBP4 detection kit.
Animals ; Baculoviridae ; genetics ; Blotting, Western ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Genetic Vectors ; Humans ; Immune Sera ; Insecta ; Rabbits ; Recombinant Proteins ; biosynthesis ; immunology ; Retinol-Binding Proteins, Plasma ; biosynthesis ; immunology ; Sf9 Cells ; metabolism ; Transfection

Heilongjiang Province has unique geographical environment, which makes the difference in population morbidity. This paper summarizes the physiological and pathological characteristics of stroke in Longjiang area from the regional factors and dietary factors. The paper also summarizes the treatment thoughts of acupuncture and moxibustion doctors in Longjiang area, mainly including five aspects: Knowing all diseases but treating the essence one, combining theory and practice, combining Chinese and western medicine, combining acupuncture and Chinese medicine, preventing diseases through learning and observing tiny clues. This article aims to summarize the experience of Longjiang acupuncture scholars and enriches the medical connotation of Longjiang area.

Objective:To explore the value of mammography-based radiomics for preoperative prediction of axillary lymph node metastasis in breast carcinoma.Methods:The clinical and X-ray data of female patients with pathologically confirmed breast cancer in Henan People′s Hospital from June 2013 to July 2017 were analyzed retrospectively. A total of 214 patients, aged 30-85 (53±11) years, were randomly divided into training set ( n=153) and verification set ( n=61) according to the ratio of 3∶1. According to pathological findings of the axillary lymph node metastasis, 99 cases were divided into positive group and 115 cases into negative group. The lesions were segmented and extracted in X-ray images of mediolateral oblique (MLO) and cranial caudal (CC). Three, nine and seven axillary lymph node metastasis related histologic features were selected from the high dimensional features of CC, MLO and CC combined MLO images by lasso regression model. According to the characteristics of imaging and clinical characteristics, the prediction model was constructed. The prediction ability of the model was verified by 10% cross validation. Results:The lymph node in positive group was larger than negative groups, the difference was statistically significant ( t=2.611, P<0.05). In the validation set, the area under curve (AUC) values of CC, MLO, CC combined with MLO images, clinical features and clinical features combined with CC and MLO images were 0.680, 0.723, 0.740, 0.558 and 0.714, respectively. Among them, CC combined with MLO images had the highest prediction efficiency, and AUC values were higher than CC alone, MLO images and CC combined with MLO images. Conclusions:Quantitative radiomics features of breast tumor extracted from digital mammograms are helpful for preoperatively predicting axillary lymph node metastasis. Future larger studies are needed to further evaluate these findings.

【Objective】 To evaluate the residual risk of hepatitis C virus (HCV) in blood screening among voluntary blood donors in Zhengzhou. 【Methods】 The ELISA and NAT screening results of 497 171 voluntary blood donors in Zhengzhou from January 2019 to December 2020 were collected through the information management system of our blood center.The residual risk of HCV was assessed using the Prevalence-Window Period Residual Risk Model. 【Results】 The residual risk among repeated and first-time blood donors was 1∶132 280 (95% CI: 1∶95 520~1∶188 820) and 1∶44 090 (95% CI: 1∶31 840~1∶62 940), respectively. The overall residual risk of blood donors screening was 1∶68 540 (95% CI: 1∶65 910~1∶130 290). The reactive rate of HCV screening in first-time blood donors (0.144%, 334/231 168) was significantly higher than that in repeated blood donors (0.014%, 36/266 003) (P<0.05), and the reactive rate of repeated blood donors in 2019 (0.019%, 26/135 267) was significantly higher than that in repeat blood donors in 2020 (0.008%, 10/130 736) (P<0.05). 【Conclusion】 The residual risk of HCV among voluntary blood donors in Zhengzhou is low.The publicity and recruitment should be further strengthened to establish a stable team of voluntary blood donation, and health consultation and physical examination should also be strengthened to further reduce the residual risk of blood transfusion.

In order to solve the pathological grading of hepatocellular carcinomas (HCC) which depends on biopsy or surgical pathology invasively, a quantitative analysis method based on radiomics signature was proposed for pathological grading of HCC in non-contrast magnetic resonance imaging (MRI) images. The MRI images were integrated to predict clinical outcomes using 328 radiomics features, quantifying tumour image intensity, shape and text, which are extracted from lesion by manual segmentation. Least absolute shrinkage and selection operator (LASSO) were used to select the most-predictive radiomics features for the pathological grading. A radiomics signature, a clinical model, and a combined model were built. The association between the radiomics signature and HCC grading was explored. This quantitative analysis method was validated in 170 consecutive patients (training dataset: = 125; validation dataset, = 45), and cross-validation with receiver operating characteristic (ROC) analysis was performed and the area under the ROC curve (AUC) was employed as the prediction metric. Through the proposed method, AUC was 0.909 in training dataset and 0.800 in validation dataset, respectively. Overall, the prediction performances by radiomics features showed statistically significant correlations with pathological grading. The results showed that radiomics signature was developed to be a significant predictor for HCC pathological grading, which may serve as a noninvasive complementary tool for clinical doctors in determining the prognosis and therapeutic strategy for HCC.
Carcinoma, Hepatocellular ; diagnostic imaging ; Humans ; Liver Neoplasms ; diagnostic imaging ; Magnetic Resonance Imaging ; Neoplasm Grading ; methods ; ROC Curve