Expression and antibody preparation of recombinant truncated glycoprotein of Guertu virus / 中华微生物学和免疫学杂志

Objective:To express truncated glycoprotein (Gn, Gn1, Gn2, Gn3, Gc1 and Gc2) of Guertu virus (GTV) in Escherichia coli ( E. coli) cells, and prepare polyclonal antibodies against recombinant proteins Gn-His, Gc1-His and Gc2-His after purification. Methods:Gene fragments encoding Gn, Gn1, Gn2, Gn3, Gc1 and Gc2 of GTV DXM strain were amplified by RT-PCR, and cloned into the prokaryotic expression vector pET-32a (+ ) to construct recombinant expression plasmids. The transformed E. coli BL21(DE3) strains carrying expression plasmids were induced by IPTG to express target proteins, which were identified by SDS-PAGE. Recombinant proteins Gn-His, Gc1-His and Gc2-His were purified by nickel affinity chromatography and detected by Western blot using GTV-positive sheep serum for analysis of their antigenicity. New Zealand white rabbits were immunized with the purified recombinant proteins. The titers and specificity of serum antibodies were analyzed by ELISA. Meanwhile, eukaryotic expression vectors pcDNA3.1-Gn, pcDNA3.1-Gc1/Gc2 were constructed and transfected into mammalian Vero cells to evaluate the binding activity of rabbit polyclonal antibodies by indirect immunofluorescence method. The specific reactivity of serum antibodies to recombinant proteins was detected by Western blot. Results:Restriction enzyme analysis and DNA sequencing confirmed that the recombinant expression vectors of pET-32a-Gn, pET-32a-Gn1/Gn2/Gn3, pET-32a-Gc1/Gc2, pcDNA3.1-Gn and pcDNA3.1-Gc1/Gc2 were constructed successfully. The relative molecular mass ( Mr) of the expressed recombinant proteins Gn-His, Gn1/Gn2/Gn3-His, Gc1/Gc2-His were approximately 63.4×10 3, 37.1×10 3, 31.9×10 3, 30.8×10 3, 40×10 3 and 54.4×10 3, respectively. The recombinant proteins could be recognized by GTV-positive sheep serum. The titers of polyclonal antibodies against GTV Gn, Gc1 and Gc2 were 1∶409 600, 1∶204 800 and 1∶6 400, respectively. Indirect immunofluorescence assay and Western blot showed that the prepared rabbit polyclonal antibodies could specifically react with the proteins expressed in eukaryotic cells and the recombinant proteins. Conclusions:The recombinant GTV glycoproteins Gn-His and Gc1/Gc2-His were efficiently expressed and purified and characterized with good immunity. The prepared polyclonal antibodies had high titers and good specificity. This study provided reference for further studying the biological function and detection methods of GTV glycoproteins and research on vaccines.

Expression and significance of serum INHB, IGF-1, VEGF and E2 in patients with polycystic ovary syndrome / 国际检验医学杂志

Objective To study the expression and significance of serum inhibin B (INHB), insulin-like growth factor-1 (IGF-1), vascular endothelial growth factor (VEGF) and estradiol (E2) in patients with polycystic ovary syndrome (PCOS).Methods A total of 45patients with PCOS in the hospital from April 2015to May 2016, were selected as observation group.In addition, 45healthy women who underwent physical examination due to male infertility were selected as control group.The significance and diagnostic value of serum INHB, IGF-1, VEGF and E2in PCOS were analyzed.Results The levels of serum INHB, IGF-1, VEGF and E2in observation group were significantly higher than those of control group (P＜0.05).The receiver operating characteristic curve (ROC curve) analysis showed that the AUC of INHB, IGF-1, VEGF and E2were 0.935, 0.988, 0.992and 0.759, respectively.The sensitivity of PCOS were 80.93％, 81.18％, 79.12％and79.07％, while the specificity of PCOS were 73.05％, 75.17％, 77.49％and 78.59％, respectively.ConclusionSerum INHB, IGF-1, VEGF and E2are highly expressed in patients with PCOS, which can be used as an important indicators in the diagnosis of PCOS.

Mediating role of resilience between stress and social anxiety in military medical university students / 第二军医大学学报

Objective To investigate the relationship between stress and social anxiety of medical students in military universities, and to explore the related mechanisms. Methods Two hundred and eleven military medical university students were tested by interaction anxiousness scale (IAS), stress scale for college students (SSCS) and Connor-Davidson resilience scale (CD-RISC). The Pearson correlation analysis and structural equation model were used to analyze the relationship between the variables. Results A total of 198 valid questionnaires were collected, with the effective rate being 93.8%. There were significant correlations between social anxiety and various dimensions of stress and resilience (all P<0.01). Structural equation model showed that the resilience partially mediated the relationship between stress and social anxiety (Χ2/df=2.08, P<0.05; root mean square error of approximation=0.074, comparative fit index=0.98, normed fit index=0.97, incremental fit index=0.98). The total stress score of the male students was greater than that of the female students in military medical universities (24.67±15.01 vs 20.17±13.07, t=2.17, P<0.05). Conclusion Resilience has partial mediating effect on the relationship between stress and social anxiety. Therefore daily management of military medical university students should be emphasized on the cultivation of resilience.

Regression Algorithm of Bone Age Estimation of Knee-joint Based on Principal Component Analysis and Support Vector Machine / 法医学杂志

Objective To establish a regression algorithm model that applies to bone age estimation of Xinjiang Uygur adolescents with machine learning methods such as histogram of oriented gradient （HOG）, local binary patterns （LBP）, support vector machine （SVM）, principal component analysis （PCA）. Methods DR images of knee-joints from 275 male and 225 female subjects aged 12.0-<19.0 years old were collected, PCA method was used to reduce the dimensionality of the HOG and LBP features, and support vector regression （SVR） was used to establish a knee-joint bone age estimation algorithm model. Stratified random sampling method was used to select 215 male samples, 180 female samples for the model training set. K-fold cross validation method was used to optimize parameters of the model. The remaining samples as the independent test set was used to compare the sample's true age and model estimated age, and had an accuracy rate in the statistical error range of ±0.8 and ±1.0 years, respectively. Then the mean absolute error （MAE） and root mean square error （RMSE） were calculated. Results The accuracy rate of male in the statistical error range of ±0.8 and ±1.0 year was 80.67%, 89.33%, respectively. The MAE and RMSE were 0.486 and 0.606 years, respectively. The accuracy rate of female in the statistical error range of ±0.8 and ±1.0 years was 80.19%, 90.45%, respectively. The MAE and RMSE were 0.485 and 0.590 years, respectively. Conclusion Establishment of prediction model for bone age estimation by feature dimension reduction of HOG and LBP in DR images of knee-joint based on PCA and SVM has relatively high accuracy.

Physical performance testing of Mobetron 2000 mobile intraoperative radiotherapy system / 医疗卫生装备

Objective To execute physical performance testing of Mobetron 2000 mobile intraoperative radiotherapy system.Methods Mobetron 2000 mobile intraoperative radiotherapy system had its indexes measured according to TG51 protocol such as percent depth dose (PDD),output dose,beam energy,field flatness and symmetry,X-ray pollution,dose linearity and repeatability.Three-dimensional water phantom was used for verification and acceptance when the measurement was executed,while the calimator end was tangent to the water surface and the nominal ource-skin distance was 50 cm.Results All the indexes of the system met the desired requirements except 12 MeV beam energy.Conclusion The system can be promoted clinically for safe,reliable and efficient treatment.

HMGB1/SREBP-1 mediated IFN-gamma-induced lipid deposition in mouse mesangial cells / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the possible mechanism of lipid deposition induced by interferon-gamma (IFN-gamma).</p><p><b>METHODS</b>The mouse mesangial cells (MMC) were randomly divided into control group, stimulation group, stimulation + control vector group (sh-HMGB1) and stimulation+ specific sh-vector group (sh-SREBP-1). RT-PCR was used to detect the expression of HMGB1, SREBP-1 and fatty acid synthetase (FAS) mRNA; the protein expression was determined by Western blot.</p><p><b>RESULTS</b>The Oil Red O staining revealed that the mouse mesangial cells showed significant lipid droplet in IFN-gamma group. IFN-gamma up-regulated the expression of HMGB1, SREBP-1, FAS mRNA and protein time-dependently; Transfection of MMC with HMGB1 siRNA resulted in the suppression of SREBP-1, FAS protein levels induced by IFN-gamma, following with decrease of lipid deposition. Stimulation with HMGB1 markedly induced expression of SREBP-1, FAS expression and peaked at 8 h, decreased at 12 h compared with that at 8 h. Sh-SREBP-1 decreased the lipid deposition induced by HMGB1 in MMC.</p><p><b>CONCLUSION</b>IFN-gamma might induce lipid deposition in mouse mesangial cells partly by up-regulating the expression of HMGB1/SREBP-1/FAS.</p>

Prevention of central pontine myelinolysis in rats by early treatment with dexamethasone / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To explore the effect and mechanism of dexamethasone (DEX) in the prevention of central pontine myelinolysis (CPM) in rats.</p><p><b>METHODS</b>Hyponatremia was induced in rat by subcutaneous injection of Vasopressin Tannate and intraperitoneal injection of 2.5% dextrose in water for 3 d, the rats of Group A received a bolus of 1 mol/L NaCl (2 ml/kg) and DEX (5 mg/kg) simultaneously at the 4th day; the rats of Group B were treated with DEX after 24 h of the injection of 1 mol/L NaCl; the rats in Group C received a bolus of 1 mol/L NaCl and saline simultaneously; Group D was the control group. The demyelinative lesions were evaluated by myelin staining. The Evans blue (EB) contents of brain were detected to evaluate the blood-brain-barrier permeability after rapid correction of hyponatremia. The expression of inducible nitric oxide synthase (iNOS) in brains was evaluated by Western blotting.</p><p><b>RESULT</b>CPM was induced successfully in rats. The EB contents of Group A, B and C had no significant difference at 0 h after injection of hypertonic saline compared with Group D. The EB contents of Group C began to increase significantly at 6 h after injection of hypertonic saline, peaked at 24 h; the expression of iNOS in brains began to increase after 3 h after the rapid correction of hyponatremia. The rate of morbidity in Group C was 66.7%. The demyelinative lesions were rarely seen in Group A, the EB contents of brain decreased significantly compared with Group C at the same time point (P<0.05), the iNOS expression was also inhibited. DEX could not prevent the attack of CPM at Group B, the rate of morbidity (75%) had no significant difference compared with Group C (P>0.05).</p><p><b>CONCLUSION</b>Early treatment with DEX can protect blood-brain-barrier and inhibit the expression of iNOS to prevent the attack of CPM.</p>

Study on tetrahydrobiopterin deficiency in Northern Chinese population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To emphasize early differential diagnosis from patients with hyperphenylalaninemia (HPA) and to evaluate the treatment and long-term outcome of patients with tetrahydrobiopterin synthase (BH4) deficiency in Northern Chinese population.</p><p><b>METHODS</b>From 1992 to 2005, a total of 618 patients with HPA were diagnosed and/or cared for in our outpatient clinic. Urinary pterin analysis, detection of dihydropteridine reductase (DHPR) activity in blood, and then BH4 loading tests were carried out to differentiate BH4 deficiency in these patients from classical phenylketonuria. BH4 deficient patients were treated with BH4, levodopa and 5-hydroxytryptophane (5-HTP) immediately while the diagnosis was done to disease. Patientso blood phenylalanine levels, psychomotor and intelligence development were followed up.</p><p><b>RESULTS</b>A total of 38 cases were diagnosed as BH4 deficiency, all of them were revealed as 6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency from the extremely decreased urine biopterin, normal DHPR activities and drop down of blood phenylalanine level to normal range within 4 to 8 hours after BH4 loading. The most common manifestations were progressively psychomotor and mental retardation to patients even after taking early dietary treatment. The patients were diagnosed and treated with drugs at the ages of 2.1 months to 13 years. With 4 patients died of pneumonia, 7 patients refused to treatment, only 27 patients were under treatment and followed up. The average full scale development or intelligence quotient (DQ/IQ) of patients who were treated within and after 6 months were 86+/- 10 or 66+/- 7 respectively. Development was not even in different aspects. A significant negative correlation was observed between the level of the DQ and the age of treatment commenced (r was -0.714, P< 0.01). Eleven patients experienced the extrapyramidal movement disorders, 3 of them combined with epilepsy. The extrapyramidal disorders were controlled by administration of levodopa.</p><p><b>CONCLUSION</b>The differential diagnosis for BH4 deficiency should be carried out in all patients with HPA. PTPS deficiency is the most common form of BH4 deficiency in Northern Chinese population. The long-term outcome of these patients benefits from diagnosis and treatment with BH4, levodopa and 5-HTP as early as possible.</p>

Epithelial growth factor receptor (EGFR) exon double-sequencing analysis in NSClC / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the mutation patterns of epithelial growth factor receptor (EGFR) exon 18, 19 and 21 in Chinese non-small-cell lung cancers (NSCLC).</p><p><b>METHODS</b>Somatic mutation in samples of 32 cases without Iressa-treatment were compared with that in 10 volunteers blood control. The mutations were identified for the forward and reverse sequence chains for the tyrosine kinase domain of the EGFR gene, followed by DNA template abstraction and Touchdown PCR.</p><p><b>RESULTS</b>Nine types of mutation were found in sequences of 7 cases among the 32 non-small cell lung carcinoma tissues, namely, five reported mutation within exon 19, and two new heterozygous mutations, L833V and H835L within exon 21, and two intron polymorphism. These results showed a mutation rate of 9/32 (28.1%) in Chinese with NSCLC, and of 31.6% in lung adenocarcinomas.</p><p><b>CONCLUSION</b>EGFR mutation rate in Chinese with NSCLC is consistent with those of Asian women reported in the literature but new mutation points in Chinese were presented as L833V and H835L. The mutation rate is in concordance with release rate of NSCLC obtained by Gefitinib treatment in Chinese.</p>

Analysis of part of M gene and genotyping for M segment of hantavirus detected from HFRS patients' sera in Qingdao region during 2000 - 2003 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the molecular epidemiological characteristics of hantavirus seen during 2000-2003 in Qingdao region of Shandong province.</p><p><b>METHODS</b>Sera were collected from 64 patients with hemorrhagic fever with renal syndrome (HFRS) and viral RNA was extracted from the sera. HTN and SEO universal primers were designed as outer primers and HTN and SEO specific primers as inner primers. G1 gene region of M segment from hantavirus was amplified by using RT-nest-PCR for sequencing. The data of nucleotide sequences were analyzed by DNA star software.</p><p><b>RESULTS</b>Six cases were positive by HTN specific primer of total cases (9%); 25 of 64 cases by SEO specific primer (39%); total positive rate was 48%. In general, SEO type was a prevalent type of hantavirus in Qingdao region. The variation of the nucleotide sequences among SEO viruses (nucleotide sequence divergence ranged from 0.3% approximately 8.9%) was lower than that among HTN type (nucleotide sequence divergence ranged from 2.6% approximately 11.2% ).</p><p><b>CONCLUSION</b>Majority of hantavirus found in Qingdao region belonged to SEO type and still a few strains belonged to HTN type. Most of the HTN viruses were detected in Jiaonan county.</p>

Antitumor effect of immunizations with fusions of dendritic and hepatocellular carcinoma cells in mice / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects of immunization with fusions of dendritic cells and H22 cells on tumor-bearing mice and their possible mechanisms.</p><p><b>METHODS</b>Fusion cells of DC and H22 cells were prepared with polyethylene glycol (PEG). Expression of MHC and costimulatory molecules by dendritomas were determined by FACs. To study the antitumor immune preventative and therapeutic effects, fusions were subcutaneously injected into tumor-bearing mice. The cytotoxic T lymphocyte (CTL) activity was determined by LDH method, the expression of TNF-a and IFN-g in tumors were assayed by RT-PCR.</p><p><b>RESULTS</b>The data showed that the hybridomas of DC and H22 cells acquired both DC and H22 cell phenotypes. Immunization of BALB/C mice with DC/H22 fusions induced potent CTL activity (mean CTL activity=0.624+/-0.024, compared with DC + H22, DC, H22 groups, F = 65.46) and a protective immunity against a high dose of H22 tumor challenge. After treatment with hybridomas, the survival time of tumor-bearing mice was greatly extended (x2=18.45). The expression levels of TNF-a and IFN-g mRNA were remarkably increased (TNF-a, F = 47.84; IFN-g, F = 37.23).</p><p><b>CONCLUSIONS</b>The hybridomas of DC and H22 cells could induce effective antitumor immune responses and may have a useful potential in prevention and management of the recurrences and metastases of HCC.</p>

Analysis on the Characteristics of Medical English Vocabulary From Derivation / 中华医学教育探索杂志

Medical English words have their own features in word formation.The medical terms are in huge sum but meanwhile of extremely typical characteristics.This paper tries to help the medical students learn these terms more efficiently by analyzing the characteristics of these terms.

Genetic characterization of the non-structural protein NSP4 from epidemic strains of human rotavirus in China / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To clarify the features of gene variation among epidemic strains of human rotavirus NSP4 in China.</p><p><b>METHODS</b>SP4 genes from 27 epidemic strains of human rotavirus isolated in different area of China in recent years were amplified with RT-PCR, the resulted cDNAs were cloned and sequenced. The sequences of full length cDNAs were compared with 10 rotavirus NSP4 sequences available in the GenBank using the Clustal x 1.8 TreeView32 and DNA Star softwares. The G serotype of VP7 was analysed by PCR.</p><p><b>RESULTS</b>The homology of the amino acid among the 27 rotavirus strains isolated in China was 81.7%-99.4%. Based on the variation of amino acid sequence, the virus strains can be divided into two groups, represented by Wa and KUN with the homology of 92.0%-99.4% and 92.0%-98.9% within each group, respectively. The diversity between the two groups were 16.6%-21.0%. The Wa group could further be separated into three subgroups, according to the diversity between those strains and the characterization in the highly variable domain. The association between VP7 serotype and NSP4 genotype was not strong.</p><p><b>CONCLUSIONS</b>The NSP4 gene of human rotavirus epidemic strains in China can be divided into Wa and KUN two groups, Wa group is the main group and contains three subgroups possessing characteristic amino acid sites. Samples isolated in the same year but not in the same area shared higher homology.</p>