Objective To explore the diagnosis value of X-ray breast imaging reporting and data system (BI-RADS) combined with wire-guided localization biopsy for breast microcalcifications in impalpable breast cancer.Methods 192 palpation negative patients with 205 microcalcification lesions were detected by mammography.All lesions were classified according to BI-RADS descriptors for calcification and were categorized by the BI-RADS.The patients with BI-RADS category 4a and above underwent X-ray positioning guide wire-guided biopsy and pathological diagnosis.Results In 205 microcalcification lesions,74 (36.1％) were malignant lesions,131 (63.9 ％) were less than benign lesions.The positive predictive value of malignant breast lesions in clustered,segmental,regional linear branching calcifications were higher [83.3 ％ (5/6),100.0 ％ (11/11),100.0 ％ (1/1)],followed by clustered,linear,segmental,regional pleomorphic calcifications [55.9 ％ (38/68),50.0 ％ (1/2),40.0 ％ (8/20) and 33.3 ％ (4/12),respectively].The positive predictive values of malignant in linear branching calcifications and pleomorphic calcifications were significantly higher than those of coarse heterogeneous calcifications,amorphous or indistinct calcifications (x2 values were 34.44,51.87,16.71,29.86,all P ＜ 0.05).The linear branching calcification had the highest possibility.The proportions of malignant lesions in four different types of glands were extremely dense 40.5 ％ (30/74),heterogeneously dense 39.2 ％ (29/74),scattered areas of fibroglandular density 10.8 ％ (8/74) and fat 9.5 ％ (7/74),respectively.Conclusions BI-RADS categorization for breast microcalcification lesions can improve the detection rate of impalpable breast cancer.Linear branching calcification has higher predictive value for malignant lesions.Dense breast is the risk factors of breast cancer,which should be attached great importance.

Objective To evaluate the clinical effect of the cannulated screw fixation in treatment of the dens fracture under locational guidance. Methods The study involved 27 patients treated with the cannulated screw fixation under locational guidance from January 2005 to January 2009.There were 19 patients with type II fracture and eight with type light m fracture.The lateral and open mouth position X-ray examination of the cervical ventebrae was done to observe the fracture healing. Results The operation lasted for a range of 40 minutes to 1.3 hour (average 1.0 hour),which showed no any complications.The patients were followed up for average 6.5 months(3-12 months),which showed sound fracture healing in 26 patients but nonhealing in one. Conclusion The cannulated screw fixation for treatment of the dens fracture under locational guidance is characterized by easy operation,minor trauma and satisfactory outcome.

ObjectiveTo evaluate the clinical effects of guiding apparatus assisted individual posterior cervical pedicle screw fixation technique.MethodsThe study enrolled 24 patients treated with posterior cervical decompressive single open-door laminoplasty and transpedicular screw-rod fixation from January 2008 to December 2010.The point of screw penetration and screw path direction were confirmed by measuring the transverse nail angle (TNA) and sagittal nail angle (SNA) of nail channel on the pre-operative CT plain scan of cervical pedicle and sagittal two-dimensional imaging of transpedicular axis.According to the results of CT measurement,individual cervical pedicle screw was implanted with the assistance of self-made guiding apparatus and then fixed after decompression.Transverse screw angle ( TSA ) and sagittal screw angle ( SSA ) were determined on the CT scan of cervical pedicle and sagittal two-dimensional imaging of transpedicular axis one week postoperatively in order to analyze the accuracy of placement of pedicle screws.Periodical anteroposterior and lateral X-ray radiographs of cervical vertebra were taken postoperatively to detect the stabilization of internal fixation.ResultsA total of 223 pedicle screws were inserted successfully into the C3 ～ C7,of which 220 crews were inserted accurately but three had slight inclination according to the postoperative CT,with placement accuracy of 98.7％.The comparison between inclination angle of inserted screws and that of preoperative transpedicular axis showed insignificant statistical difference ( P ＞ 0.05 ).All the patients were followed up for 6-34 months ( mean,18.5 months),which showed no neurovascular complications related to screws perforation out of pedicle cortex or no screw loosening,prolapse or breakage.ConclusionPosterior cervical pedicle screw insertionperformed according to the individual CT measurement is easy and safe and has a high accuracy rate under the assistance of self-made guiding apparatus.

Objective To investigate the humoral and cellular immune responses in patients with acute brucellosis, and evaluate dynamic changes of regulatory T-lymphocytes (Foxp3+ Treg) in the peripheral blood of patients during treatment, in order to clarify the relationship between immunosuppression and the therapeutic effect in human brucellosis.Methods Sixty-five patients with brucellosis hospitalized at the Third Department of Infectious Diseases, Heilongjiang Agriculture and Reclamation Bureau General Hospital between July 2015 and November 2015 were included.Twenty-eight patients were treated with conventional therapy (group A: patients received 3 courses of treatment.Each lasted for 20 days with one-week interval), and 37 patients were treated with conventional therapy in combination with immunopotentiator (group B).Thirty healthy volunteers were enrolled as the controlled group.The ratio of CD3+CD4+ Foxp3+ Treg cells in the peripheral blood of brucellosis patients were measured by flow cytometry (FCM) at the end of each course of treatment.Data in accordance with normal distribution were described as mean±standard deviation.Comparison between two groups was done by two sample t test.Comparison among multiple groups was performed by analysis of variance and SNK test.Data that did not fit the normal distribution were analyzed by multiple-sample nonparametric test.Results After the first (20 d), second (50 d) and third course of treatment (80 d), the ratios of Foxp3+Treg in the peripheral blood of 65 acute brucellosis patients were 2.83%, 3.77% and 4.03%, respectively, which were all significantly higher than control group (1.69%;t=5.97, 9.05 and 5.66, respectively, all P<0.01).At the end of the first course of treatment, the ratios of Foxp3+Treg in group A and B showed no statistically difference (t=0.33, P>0.05), while those were both higher than control group (t=7.09 and 4.94, respectively;both P<0.01).At the end of the second course, the ratio of Foxp3+ Treg in group B was higher than group A (t=2.22, P<0.01), and both of them were higher than control group (t=10.79 and 7.25, respectively;both P<0.01).At the end of treatment, Foxp3+ Treg in group A was also significantly higher than the other two groups (t=6.02 and 6.45, respectively;both P<0.01).Conclusions In patients with acute brucellosis treated with the standard antibiosis treatment in combination with immunopotentiator, the ratio of Foxp3+Tregs significantly increases and maintains at a high level, which suggests that extra immunopotentiator may be not helpful for the treatment of brucellosis at the very early stage.

Maize is one of the most important food crops. Rice black-streaked dwarf virus is a maize rough dwarf disease pathogen. The occurrence and transmission of maize rough dwarf disease brings great damage to maize production. The technology of using artificial miRNA to build antiviral plant has been proven effective in a variety of plants. However, such trials in maize have not been reported. We designed primers based on the sequence of maize zea-miR159a precursor and sequence of function protein genes and silencing RBSDV coding genes in RBSDV genome. We constructed amiRNA (artificial miRNA) gene for silencing RBSDV coding gene and gene silencing suppressor. We constructed pCAMBIA3301-121-amiRNA plant expression vector for transforming maize inbred lines Z31 by using agrobacterium mediated method. After molecular analysis of transgenic maize, homozygous lines with high miRNA expression were selected by molecular detection for a subsequent natural infection experiment. We studied the severity of maize rough dwarf disease according to a grading standard (grade 0 to 4). The experiment results showed that the disease resistance of transgenic homozygous maize with the anti-rough dwarf virus amiRNA vector was better than that of wild type. Among the transgenic maize, S6-miR159 transgenic maize had high disease resistance. It is feasible to create new maize variety by the use of artificial miRNA.
Disease Resistance ; genetics ; Gene Silencing ; Genetic Vectors ; MicroRNAs ; genetics ; Plant Diseases ; genetics ; virology ; Plants, Genetically Modified ; genetics ; Reoviridae ; pathogenicity ; Zea mays ; genetics

Objective To investigate the percentage of regulatory T cells (Treg) in peripheral blood lymphocytes of patients with chronic brucellosis and the percentage change before and after treatment of different regimens,and to analyze the influence of Treg cell-induced immunosuppression on the therapeutic effect of chronic stage brucellosis.Methods Using case-control study,35 patients with chronic brucellosis who were hospitalized in Heilongjiang General Hospital of Agriculture Bureau [28 males,7 females,aged (45.37 ± 20.16) years old] were selected as case group.According to the treatment regimen,they were divided into standard treatment group (15 cases) and immune enhancer group (20 cases),the treatment was 20 d;30 cases of in-hospital health examinations were selected [16 males and 14 females,aged (35.53 ± 11.38) years old] as control group.Peripheral blood sample of the subject was collected before and after the treatment,the Treg cells as a percentage in peripheral blood lymphocytes were detected by flow cytometry.And the percentage change of Treg cells of brucellosis patients who underwent different treatment regimens was analyzed.Results Before treatment,the percentage of Treg cells in peripheral blood lymphocytes of the control group,the standard therapy and the immune enhancer groups [(1.69 ± 0.38)％,(3.12 ± 0.86)％,(3.05 ± 1.07)％] was significantly different (F =25.89,P ＜ 0.05);compared with the control group,the percentage of Treg cells in the peripheral blood lymphocytes of the standard treatment group and the immune enhancer group increased (P ＜ 0.05);there was no significant difference between the standard treatment group and the immune enhancer group (P ＞ 0.05).After treatment,the percentage of Treg cells in peripheral blood lymphocytes of the control group,the standard therapy and the immune enhancer groups [(1.69 ± 0.38)％,(3.06 ± 0.76)％,(2.85 ± 0.89)％] was significantly different (F =30.84,P ＜ 0.05);compared with the control group,the percentage of Treg cells in the peripheral blood lymphocytes of the standard treatment group and the immune enhancer group increased (P ＜ 0.05);there was no significant difference between the standard treatment group and the immune enhancer group (P ＞ 0.05),and compared with the same group before the treatment,respectively,the differences were not statistically significant (P ＞ 0.05).Conclusions The percentages of Treg cells in peripheral blood lymphocytes of the chronic brucellosis patient are not significantly changed before and after different treatment regimens.It suggests that the immunesuppression induced by Treg cells may be one of the reasons why the host organism cannot effectively remove residual Brucella in the body,which leads to chronic infection.

Objective: To assess the effect of viral macrophage inflammatory protein (vMIP) -Ⅱ on the expression of apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (APOBEC3G) , and to explore the mechanisms. Methods: A recombinant plasmid pEGFP-N3-K4 (vMIP-Ⅱ plasmid group) and an empty plasmid pEGFP-N3 (empty plasmid group) were separately transfected into 293T cells, and quantitative PCR and Western blot analysis were performed to evaluate the effect of transfection with vMIP-Ⅱ gene on the APOBEC3G expression in 293T cells. Some 293T cells in the empty plasmid group and vMIP-Ⅱ plasmid group were treated with 1 000 IU/ml interferon (IFN) -α for 36 hours, and then Western blot analysis was conducted to determine the APOBEC3G expression in the empty plasmid group and vMIP-Ⅱ plasmid group with or without IFN-α treatment. Some 293T cells transfected with vMIP-Ⅱ plasmids were treated with 75 μmol/L AG490 (a JAK/STAT signaling pathway inhibitor) and 20 μmol/L U0126 (an ERK signaling pathway inhibitor) separately; after 24 hours, total protein was extracted from 293T cells, and Western blot analysis was conducted to determine the expression of APOBEC3G. A recombinant plasmid containing APOBEC3G promoter was constructed by using a luciferase reporter gene, and the promoter fragment included the full-length promoter sequence (POS) of APOBEC3G, sequences with the lengths of 1 560, 960, 720, 480, 420, 360, 330 and 240 bp, and the regulatory element-free region (NEG) of APOBEC3G, separately. Some 293T cells were co-transfected with the recombinant plasmid carrying luciferase reporter gene and vMIP-Ⅱ plasmid (experimental group), or the recombinant plasmid and empty plasmid (control group). Subsequently, the activity of the APOBEC3G promoter was evaluated, and the key promoter region through which the transcriptional activity of APOBEC3G was regulated by vMIP-Ⅱ was analyzed. Statistical analysis was carried out by using t test, one-way analysis of variance and least significant difference (LSD) -t test. Results: The mRNA and protein expression of APOBEC3G was significantly higher in the vMIP-Ⅱ plasmid group (2.500 ± 0.013, 1.472 ± 0.013 respectively) than in the control group (1, 0.364 ± 0.030 respectively; t = 6.22, 6.54 respectively, both P < 0.05) . The APOBEC3G expression significantly differed among the empty plasmid group, vMIP-Ⅱ plasmid group, empty plasmid + IFN-α group and vMIP-Ⅱ plasmid + IFN-α group (1, 2.030 ± 0.108, 2.700 ± 0.081 and 2.600 ± 0.099 respectively; F = 67.026, P < 0.001) , but there was no significant difference between the vMIP-Ⅱ plasmid group and empty plasmid + IFN-α group (t = 3.46, P > 0.05) . The APOBEC3G expression also significantly differed among the vMIP-Ⅱ plasmid group, vMIP-Ⅱ plasmid + AG490 group and vMIP-Ⅱ plasmid + U0126 group (0.617 ± 0.025, 0.179 ± 0.061, 0.359 ± 0.012 respectively; F = 70.019, P < 0.001) , and was significantly lower in the vMIP-Ⅱ plasmid + AG490 group and vMIP-Ⅱ plasmid + U0126 group than in the vMIP-Ⅱ plasmid group (t = 9.66, 11.836 respectively, both P < 0.01) . Luciferase activity assay showed that the promoter activity significantly differed among the vMIP-Ⅱ plasmid groups transfected with POS, 1 560-, 960-, 720-, 480-, 420-, 360-, 330-, 240-bp or NEG sequences (F = 81.092, P < 0.001) , and the APOBEC3G promoter activity decreased greatly from the 720-bp group to 480-bp group. Conclusion vMIP-Ⅱ upregulates the expression of APOBEC3G, likely through the JAK/STAT signaling pathway or the key promoter region regulating the transcriptional activity of APOBEC3G.

Photosynthetic CO(2) fixation is the ultimate source of organic carbon on earth and thus is essential for crop production and carbon sequestration. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the first step of photosynthetic CO(2) fixation. However, the extreme low carboxylation efficiency of Rubisco makes it the most attractive target for improving photosynthetic efficiency. Extensive studies have focused on re-engineering a more efficient enzyme, but the effort has been impeded by the limited understanding of its structure-function relationships and the lack of an efficient selection system towards its activity. To address the unsuccessful molecular engineering of Rubisco, we developed an Escherichia coli-based activity-directed selection system which links the growth of host cell solely to the Rubisco activity therein. A Synechococcus sp. PCC7002 Rubisco mutant with E49V and D82G substitutions in the small subunit was selected from a total of 15,000 mutants by one round of evolution. This mutant showed an 85% increase in specific carboxylation activity and a 45% improvement in catalytic efficiency towards CO(2). The small-subunit E49V mutation was speculated to influence holoenzyme catalysis through interaction with the large-subunit Q225. This interaction is conserved among various Rubisco from higher plants and Chlamydomonas reinhardtii. Knowledge of these might provide clues for engineering Rubisco from higher plants, with the potential of increasing the crop yield.
Amino Acid Substitution ; Bacterial Proteins ; chemistry ; genetics ; Carbon Dioxide ; chemistry ; Directed Molecular Evolution ; Escherichia coli ; growth & development ; Ribulose-Bisphosphate Carboxylase ; chemistry ; genetics ; Synechococcus ; enzymology

Objective:To study the correlation between serum bilirubin and cystatin C in patients with type 2 diabetes mellitus.Methods:A retrospective cohort study was conducted on 750 patients who were in the Affiliated Hospital of Jining Medical University from June 2017 to May 2018. The clinical data were collected, and the correlation between serum total bilirubin, direct bilirubin, indirect bilirubin and cystatin C was analyzed.Results:According to the results of single factor analysis, after adjusting the related confounding factors, the smooth curve fitting showed that there was a U-shaped relationship between the total bilirubin, indirect bilirubin and cystatin C. When the total bilirubin was <15.9 μmol/L, for every increase of 1 μmol/L in total bilirubin, cystatin C decreased 0.008 mg/L ( β = - 0.008, 95% CI - 0.014 to - 0.002, P<0.01); when indirect bilirubin was <11.5 μmol/L, for every increase of 1 μmol/L in indirect bilirubin, cystatin C decreased 0.011 mg/L ( β = - 0.011, 95% CI - 0.018 to - 0.003, P<0.01). When cystatin C was grouped according to the normal range (cystatin C<1.25 mg/L), after adjusting the related confounding factors, the smooth curve fitting showed that there was a U-shaped relationship between the total bilirubin and indirect bilirubin with cystatin C; when total bilirubin was <15.5 mol/L, for every increase of 1 μmol/L in total bilirubin, the risk of cystatin C exceeding the normal value was reduced by 17% ( OR = 0.83, 95% CI 0.71 to 0.96, P<0.01); when total bilirubin was ≥15.5 μmol/L, for every increase of 1 μmol/L in total bilirubin, the risk of cystatin C exceeding the normal value was increased by 12% ( OR = 1.12, 95% CI 1.01 to 1.25, P<0.05); when indirect bilirubin was <11.8 μmol/L, every increase of 1 μmol/L in indirect bilirubin, the risk of cystatin C exceeding the normal value was reduced by 20% ( OR = 0.80, 95% CI 0.67 to 0.95, P<0.01). However there was no significant correlation between direct bilirubin and cystatin C. Conclusions:There is a U-shaped relationship between total bilirubin, indirect bilirubin and cystatin C. At physiological concentrations, the increase of total bilirubin and indirect bilirubin can reduce cystatin C.

The clinical data of a case with late-onset isolated sulfite oxidase deficiency(ISOD)admitted in the Department of Neurology, Children′s Hospital, Zhejiang University School of Medicine in July 2021 were retrospectively analyzed.Fifteen previously published cases of late-onset ISOD were also reviewed.The patient was a girl, who was hospitalized because of " motor regression with mental retardation for 5 days" at 1 year old.The manifestations of the patient were extrapyramidal symptoms, regression of motor development and seizures.The level of urinary sulfites in the patient was increased.Magnetic resonance imaging (MRI) features were bilateral pallidus and substantia nigra.Gene sequencing suggested a pure missense mutation of the sulfite oxidase( SUOX) gene c. 650(exon5)G>A(p.Arg217Gln). In 16 cases of late-onset ISOD, the median age at onset and diagnosis was 10.5 months and 34.0 months, respectively.The common clinical manifestations were hypotonia (13 cases), seizures (10 cases), movement disorders (9 cases), and ectopia lentis (6 cases). The most common brain MRI feature was pallidus changes (11 cases), followed by lesions of substantia nigra (5 cases), and cerebral atrophy (4 cases). Fourteen cases of late-onset ISOD showed a positive urinary sulfite test.The missense mutation of the SUOX gene was found in 9 cases.It suggested that brain MRI involvement of bilateral pallidus, high excretion of urine sulfites and the missense mutation of the SUOX gene were important diagnostic clues for late-onset ISOD.