Incidence of unintended pregnancy within 2 years after delivery and its influencing factors in China / 中华妇产科杂志

Objective:To investigate the present situation of unintended pregnancy within two years postpartum and its influencing factors in China.Methods:Participants who delivered a live birth at 60 hospitals in 15 provinces in the eastern, central and western regions of China during July 2015 to June 2016 were interviewed by using structured questionnaire. Information on occurrence of unintended pregnancy within 2 years after delivery, postpartum contraceptive use, sexual resumption, breastfeeding, and women′s socio-demographic characteristics, and so on, were collected. Life-table analysis, cluster log-rank tests and a 2-level Cox regression model were used for data analysis.Results:A total of 18 045 postpartum women were investigated. The cumulative 1- and 2-year unintended pregnancy rates after delivery were 5.3% (95% CI: 4.5%－6.1%) and 13.1% (95% CI: 11.3%－14.8%), respectively. Cox regression model analysis showed that the risk of unintended pregnancy within 2 years postpartum were increased in younger women, ethnic minorities, women with abortion history, and those who had a vaginal delivery with short lactation time and late postpartum contraceptive initiation (all P<0.01). The risk of postpartum unintended pregnancy was not associated with geographic regions and hospitals where women gave a birth (all P>0.05). Conclusions:In China, the risk of unintended pregnancy within 2 years after delivery is relatively high. Service institutions and service providers should improve the quality of postpartum family planning services, promote the use of high effect contraceptive methods, and educate women to use a method at the time of their sexual resumption or even before.

Prognostic factors for failure of transvaginal repair of vesicovaginal fistula: A nested case-control study / 北京大学学报(医学版)

OBJECTIVE@#To analyze the prognostic factors affecting the failure of transvaginal repair of vesicovaginal fistula (VVF).@*METHODS@#A retrospective nested case-control study was conducted. A total of 15 patients who underwent unsuccessful transvaginal vesicovaginal fistula repair in the Department of Urology, Peking University First Hospital from January 2014 to December 2020 were enrolled as the case group. A total of 60 patients receiving transvaginal vesicovaginal fistula repair by the same surgeon within the same time range, were selected as the control group. The age, body mass index (BMI), etiology of vesicovaginal fistula, associated genitourinary malformation, frequency of repair, characteristics of fistula, surgical procedure, postoperative recovery and other factors were compared between the case group and the control group, and the influencing factors of failure were analyzed.@*RESULTS@#The BMI of the case group was (26.3±3.9) kg/m2, the diameter of vaginal fistula was (1.5±0.8) cm, and the operative time of transvaginal repair was (111.8±19.8) min. The proportion of the patients with genitourinary malformations was 4/15, the proportion of the patients with multiple vaginal repairs was 13/15, the proportion of the patients with concurrent ureteral reimplantation was 6/15, and the proportion of the patients with postoperative fever was 5/15. In the control group, the BMI was (23.9±3.0) kg/m2, the diameter of vaginal fistula was (0.8±0.5) cm, the operative time of transvaginal repair was (99.9±19.7) min, the rate of associated genitourinary malformation was 2/60, the rate of multiple transvaginal repair was 18/60, the rate of concurrent ureteral reimplantation was 5/60, and no postoperative fever was found. Compared with the control group, the case group had higher BMI (P=0.013), bigger vaginal fistula (P=0.002), longer time of operation (P=0.027), higher proportion of genitourinary malformations (P=0.013), higher proportion of repeated transvaginal repair (P < 0.001), higher proportion of ureter reimplantation (P=0.006), and higher proportion of postoperative fever (P < 0.001). Multivariate analysis showed that fistula diameter ≥1 cm (OR=10.45, 95%CI=1.90-57.56, P=0.007) and repeated transvaginal repair (OR=16.97, 95%CI=3.17-90.91, P=0.001) were independent prognostic factors for VVF failure in transvaginal repair.@*CONCLUSION@#Fistula diameter ≥1 cm and repeated transvaginal repair are independent prognostic factors of failure in transvaginal repair.

Investigation and analysis of cesarean section rate and indications in Shanghai / 中国实用妇科与产科杂志

OBJECTIVE:To study the current status of cesarean section rate(CSR)and cesarean section(CS)indications in Shanghai,and to provide basic evidence for reducing the rate of CS.METHODS:Six general and maternal and child hospi⁃tals(MCH)in Shanghai were selected,and all data of childbirth of these hospitals in 2016 were collected.The total CSR was calculated for all hospitals and the Chi-square tests were used to compare the CSR among different hospitals.The constitution of CS indications were reported by using the data collected from two out of the six hospitals.RESULTS:The to⁃tal CSR of all hospitals in this study was 47.88%(95% CI 47.30%-48.46%).The CSR in suburb an hospitals was signifi⁃cantly higher than that in outer suburbs and city center(51.72% vs. 50.54% vs. 43.24%,P<0.01),The CSR in secondary hospitals was significantly higher than that in tertiary hospitals(49.73% vs. 46.36%,P<0.01).The CSR in general hospi⁃tals was higher than that in MCH(54.54% vs. 43.81%,P<0.01).The CS without medical indications or for social reasons accounted for 25.15% of all CS,while the scar uterus(36.31%),maternal pregnancy-related complications(13.17%)and abnormal fetal head position(6.33%)were the three leading medical indications of CS.CONCLUSION:The CSR in Shang⁃hai is at a high level.There are significant differences in CSR among different hospitals.The scar uterine and social rea⁃sons are the leading causes of CS in Shanghai.

A survey of risk factors and quality of life in female medical staff with urinary incontinence / 北京大学学报(医学版)

OBJECTIVE@#To explore risk factors of urinary incontinence (UI) and influences of UI on quality of life in female medical staff from Beijing.@*METHODS@#One hundred and forty-six female medical personnel were included in the present study through the convenient sampling method in Peking University First Hospital. The questionnaires contained the following information: demographic information, daily urination condition, the severity of UI [international consultation on incontinence questionnaire short form (ICI-Q-SF)], and the influences of UI on quality of life (QOL). We excluded the subjects who were in pregnancy or had urinary infection, neurogenic bladder, or urethral stricture. We used SPSS 21.0 software (IBM Corp, Armonk, NY) for statistical analysis. The Kolmogorov-Smirnov test determined the normality of the continuous variables. Means and standard deviation presented continuous variables in normality. Median and range presented continuous variables without normality. Frequency and percentile were used to present categorical or ranked variables.@*RESULTS@#There were 63 out of 146 (43.2%) female medical staff suffering from UI. The mean age and body mass index of the whole study cohort were (39.4±9.9) years and (22.3±3.4) kg/m2. The median delivery times of all the subjects were 1 time (range: 0-3 times). Fifty out of 146 (34.2%) subjects had transvaginal delivery history. Chronic constipation was diagnosed in 32 subjects (21.9%). No significant difference in daytime micturition and nocturia times were found between the UI and non-UI subjects. According to the multivariate analysis, chronic constipation (OR=4.95, 95%CI=1.81-13.53, P=0.002) and transvaginal delivery history (OR=3.50, 95%CI=1.49-8.21, P=0.004) were independent risk factors for UI. The non-UI subjects had superior quality of life than the UI subjects in terms of incontinence quality of life (I-QOL) total scores and all sub-scores of avoidance and limiting behaviors, psychosocial impacts, and social embarrassment (P<0.001). In addition, avoidance and limiting behaviors (r=-0.449, P<0.001), psychosocial impacts (r=-0.538, P<0.001), and social embarrassment (r=-0.454, P<0.001) of the 63 UI subjects were equally influenced by the incontinence symptom.@*CONCLUSION@#UI is not uncommon in female medical staff. The quality of life of medical faculty is influenced by UI in terms of avoidance and limiting behaviors, psychosocial impacts, and social embarrassment. Chronic constipation and transvaginal delivery history were independent risk factors for UI.

Research advances in the role of polyamine metabolism in aging and heart failure / 中华老年医学杂志

The incidence of aging-induced heart failure has increased.Spermidine,spermine,and putrescine,the intermediate metabolites of polyamine metabolism,not only participate in an array of crucial molecular interactions with DNA,RNA,and proteins involved in gene transcription and cell division,but also change with aging and age-associated cardiac dysfunction.Detection of serum polyamine metabolites and B type natriuretic peptide(BNP)may open up new ways for the assessment of aging and aging-induced cardiac dysfunction.

Ultrasonographic Evaluation of Infundibular Bladder Neck Formation in Female Stress Urinary Incontinence / 中国医学影像学杂志

Purpose To investigate the relationship of infundibular bladder neck formation with pelvic floor support structure injury and urethral sphincter defect and its significance in female stress urinary incontinence.Materials and Methods The pelvic floor images of seventy-four female patients with stress urinary incontinence treated in the outpatient Department of Lanzhou University Second Hospital from April 2015 to August 2016 were analyzed retrospectively.The location of the bladder neck,posterior vesicourethral angle and the infundibular bladder neck formation were observed by the transperineal ultrasound under the resting state and the maximum Valsalva status.Meanwhile the thickness of middle urethral sphincter was measured under resting state.At the same time,eighty-one women visiting our hospital for regular physical examination were enrolled as control group.Results The infundibular urinary bladder neck formation rate (66.2％) in the stress urinary incontinence group was significantly higher than that in the control group (4.9％) under maximum Valsalva state,the difference was statistically significant (P＜0.05).The extent of the bladder neck descending and posterior vesicourethral angle in the stress urinary incontinence group were notably higher than those in the control group,both of the difference was statistically significant (P＜0.05).Stress urinary incontinence was confirmed with urethral sphincter defect by urodynamics in nine patients,in whom the infundibular bladder neck occurred.The thickness of the middle urethral sphincter in these nine patients showed no obvious difference with that in patients without sphincter defect and subjects in normal control group (P＞0.05).Conclusion The infundibular bladder neck formation,which is closely related to the pelvic floor support structure dysfunction and urethral sphincter defect,is an important indication of stress urinary incontinence.However,the assessment of urethral sphincter defect through urethral sphincter thickness need to be further studied.

Research on Glycyrrhizeae Radix standard decoction / 中国中药杂志

Fifteen batches of Glycyrrhizeae Radix standard decoction were prepared for determination of the content of the glycyrrhizic acid and liquiritin, then the transfer rate and the extract rate were calculated and a method was established to analyze the fingerprint by HPLC. According to the measurement of 15 batches of samples,the transfer rate of the glycyrrhizic acid and liquiritin were 59.4%-87.4% and 49.8%-78.9% with extract rate of 29.9%-38.9%. Moreover,10 common chromatographic peaks were determined based on their fingerprint by using similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (TCM)(2012A) .The similarity results of 15 batches of samples were analyzed and compared,and the results showed that the similarity was all higher than 0.9. Fifteen batches of samples,prepared by a standard method,have stable quality and the high similarity.The method displayed good precision,stability and repeatability in fingerprint analysis. Therefore,this study can provide a reference for the quality control of Glycyrrhizae Radix dispensing granules.

Application of fast-track surgery concept in the perioperative nursing care of andrological patients: A randomized controlled study / 中华男科学杂志

<p><b>Objective</b>To evaluate the application of the fast-track surgery (FTS) concept in the nursing care of andrological patients during the perioperative period.</p><p><b>METHODS</b>A total of 200 males to be treated by andrological surgery were included in a control group and another 200 in an observation group, the former received conventional perioperative nursing care, while the latter underwent an FTS nursing care procedure including a variety of proven effective methods to reduce surgical stress and achieve a quick recovery during the perioperative period. Comparisons were made between the two groups of patients in the postoperative enterokinesia time, anal exhaust time, eating time, off-bed time, defecating time, bowel preparation complications, and degree of comfort and satisfaction.</p><p><b>RESULTS</b>Compared with the controls, the patients in the observation group showed significantly earlier postoperative enterokinesia time (［5.8±0.9］ vs ［4.4±1.4］ h, P<0.01), anal exhaust time (［10.8±1.8］ vs ［7.7±2.0］ h, P<0.01), eating time (［12.9±0.7］ vs ［6.3±0.7］ h, P<0.01), off-bed time ［14.3±2.7］ vs ［8.2±1.4］ h, P<0.01), and defecating time (［49.2±2.6］ vs ［39.6±2.5］ h, P<0.01), a lower incidence of bowel preparation complications (P<0.01), and a higher degree of comfort (P<0.01) and satisfaction (［97.5±0.7］% vs ［99.4±+0.3］ %, P<0.01).</p><p><b>CONCLUSIONS</b>The FTS concept can be safely and effectively applied to the perioperative nursing care of andrological patients to achieve a faster recovery and higher degree of comfort and satisfaction postoperatively.</p>

Aging-related change of erectile function and the expression of SIRT1 / 中华男科学杂志

<p><b>Objective</b>To explore aging-related changes in erectile function and the expressions of SIRT1 and other relevant factors in rats.</p><p><b>METHODS</b>We divided 40 male SD rats into four age groups of equal number: 2-month-old (2 mo), 8-month-old (8 mo), 14-month-old (14 mo), and 20-month-old (20 mo), measured the intracavernous pressure (ICP), mean arterial pressure (MAP), and ICP/MAP ratio by electrostimulation of the cavernous nerve, evaluated fibrosis in the corpus cavernosum by Masson's trichrome staining, detected the expressions of SIRT1, P53, and FOXO3a by Western blot, and determined the levels of NO and cGMP using the NO/cGMP kit.</p><p><b>RESULTS</b>Both the ICP/MAP ratio and the cGMP level were elevated with aging, reaching the peak at 8 months and then gradually decreased. Masson staining showed an aging-related increase of collagen fibers in the corpus cavernosum.The expression of SIRT1 was reduced while those of P53 and FOXO3a increased with aging.</p><p><b>CONCLUSIONS</b>Aging-related erectile dysfunction may be attributed to the reduced activity of the NO/cGMP pathway, apoptosis and oxidative stress, and SIRT1 may play a role in aging-related erectile dysfunction.</p>

Combination of acupressure and magnetic sticker improved the quality of life in patients with advanced gastroenteric tumor: a clinical observation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the clinical effect of combination of acupressure and magnetic sticker on the quality of life (QOL) including appetite, defecation, and sleep in patients with advanced gastroenteric tumor.</p><p><b>METHODS</b>Totally 147 patients with advanced gastroenteric tumor were assigned to 4 groups according to different treatment methods, i.e., the supportive treatment group (A, 20 cases), the acupressure treatment group (B, 41 cases), the magnetic sticker treatment group (C, 40 cases), and a combination of acupressure and magnetic sticker treatment group (D, 46 cases). They were respectively treated with different methods, supportive treatment for group A, acupressure for group B, magnetic sticker for group C, and a combination of acupressure and magnetic sticker for group D. The scores of food intake, defecation frequency, sleep time, Karnofsky, and QOL were compared before treatment and at day 14 after treatment.</p><p><b>RESULTS</b>After treatment, the scores of food intake, defecation frequency, and sleep time were obviously improved in B, C and D groups (P < 0.01). There was statistical difference between group D and group A (P < 0.01). In addition, in comparison with A group, both Karnofsky score and QOL score increased in B, C and D groups (P < 0.01).</p><p><b>CONCLUSION</b>The assisted therapy of the combination of acupressure and magnetic sticker could ameliorate QOL such as the digestive functions and sleep in patients with advanced gastroenteric tumor.</p>

Establishment and preliminary application of dengue virus envelope domain III IgG antibody capture enzyme-linked immuno-absorbent assay / 中华预防医学杂志

<p><b>OBJECTIVE</b>To establish a highly sensitive and specific assay to detect dengue virus (DENV) envelope protein domain III (EDIII) IgG antibody, and to explore its value in the diagnosis and seroepidemiological survey of dengue.</p><p><b>METHODS</b>The DENV EDIII IgG antibody capture ELISA was developed using the recombinant full-length DENV EDIII, which was prepared by Pichia yeast expression system as the capture antigen. The serum samples were collected from the same group of 35 DENV-1 patients of primary infection during disease period in 2006 and their follow-up phase in 2010; and the sensitivity of the assay was compared to that of the commercial Panbio DENV IgG ELISA.</p><p><b>RESULTS</b>The sensitivity of DENV EDIII IgG ELISA in detecting the serum samples from disease period and follow-up phase was 87% (20/23) and 94% (33/35), respectively; whereas the sensitivity of Panbio DENV IgG ELISA was 71% (25/35) and 0, respectively. The sensitivity of DENV EDIII IgG ELISA in detecting the serum samples from both periods was similar, without statistical significance (χ(2) = 0.946, P = 0.331). For serum samples from disease period, the sensitivity of DENV EDIII IgG ELISA was comparable with that of Panbio DENV IgG ELISA (χ(2) = 1.924, P = 0.165). However, DENV EDIII IgG ELISA demonstrated a significantly higher sensitivity than Panbio DENV IgG ELISA in detecting the serum samples from follow-up phase (χ(2) = 62.432, P = 0.000).</p><p><b>CONCLUSION</b>DENV EDIII IgG capture ELISA is highly sensitive in detecting IgG in the serum samples from either disease period or follow-up phase. This method might be a promising alternative for diagnosis and seroepidemiologic survey of dengue.</p>

Gene cloning and immunogenicity analysis of the structural proteins VP1-VP4 of enterovirus 71 / 南方医科大学学报

<p><b>OBJECTIVE</b>To clone the genes encoding the structural proteins VP1-VP4 of enterovirus 71 and investigate the immunogenicity of the expressed recombinant proteins.</p><p><b>METHODS</b>The VP1-VP4 cDNAs were amplified by RT-PCR from the extracted viral RNA and cloned into pMD19-T vector. The cloned VP1-VP4 genes were then inserted into the multi-cloning sites of plasmid pQE30a and expressed in E. coli M15 with IPTG induction. After washing with 8 mol/L urea and purification with Ni-affinity chromatography, the recombinant proteins obtained were tested for immunogenicity by Western blotting and ELISA using rabbit antisera against enterovirus 71 and Coxsackie Virus A16.</p><p><b>RESULTS</b>The recombinant VP1-VP4 proteins were highly expressed in E. coli M15 and the purified proteins could be specifically recognized by the rabbit sera against enterovirus 71.</p><p><b>CONCLUSION</b>The expressed enterovirus 71 structural proteins show good immunogenicity and can be used for developing enterovirus 71 vaccine and detection kits.</p>

Characterization of specific monoclonal antibodies to Aspergillus conidia by flow cytometry / 南方医科大学学报

<p><b>OBJECTIVE</b>To characterize the specific monoclonal antibodies to Aspergillus conidia.</p><p><b>METHODS</b>Flow cytometry was used to examine the reactivity of the specific monoclonal antibodies to Aspergillus conidia.</p><p><b>RESULTS</b>Both the monoclonal antibodies MA3 and Con2 showed specific reactivity to Aspergillus conidia suspensions. MA3 was capable of binding to the conidia of A.fumigatus, A.flavus, A.niger and A.terreus, while Con2 was reactive only to the conidia of A.fumigatus.</p><p><b>CONCLUSION</b>Two specific monoclonal antibodies (MA3 and Con2) to Aspergillus conidia have been obtained.</p>

Characterization and secreted expression of dengue virus type I-IV envelope glycoprotein domain III in Pichia pastoris / 中华预防医学杂志

<p><b>OBJECTIVE</b>To achieve secretory and extracellular production of recombinant dengue virus serotypes I-IV envelope glycoprotein domain III (DENV-1-4 EDIII) in Pichia pastoris.</p><p><b>METHODS</b>EDIII genes of DENVI-IV were amplified and cloned into vector pPIC9K, respectively. These recombinant plasmids were then linearized and transferred into Pichia pastoris strain GS115. Clones highly produced in 4.0 mg/ml G418 were amplified and induced by methanol to achieve the secreted recombinant proteins. Ni-NTA agarose beads were used for purification, while SDS-PAGE and Western blotting were used for identification.</p><p><b>RESULTS</b>The recombinant plasmids pPIC9K-DENV-1-4 EDIII were constructed and successfully transferred into Pichia pastoris strain GS115. The recombinant EDIII proteins were expressed in a secretory way with the molecular weight about 12 × 10(3) and specifically identified by anti-His monoclonal antibody and anti-DENVI-IV mice sera.</p><p><b>CONCLUSION</b>DENVI-IV EDIII proteins are successfully achieved from Pichia pastoris expression system and could be used for development of dengue vaccines, diagnostic reagents and study of biological function of the E protein.</p>

To evaluate the neutralizing abilities of anti-dengue virus antibodies with nonstructural protein 1 antigen capture enzyme-linked immunosorbent assay / 中华预防医学杂志

<p><b>OBJECTIVE</b>To produce neutralizing antibodies against envelope protein domain III (EDIII) of dengue virus serotype I (DENV-1) and evaluate the nonstructural protein 1 (NS1) antigen capture enzyme-linked immunosorbent assay (ELISA) for identification of antibody neutralizing abilities.</p><p><b>METHODS</b>Five BALB/c mice and one New Zealand Rabbit were immunized with recombinant EDIII protein of DENV-1 for the production of hybridomas and hyperimmune sera. Indirect ELISA, immunofluorescence assay (IFA) and Western Blot analyses were applied to identify specificity of antibodies. Comparing to plaque reduction neutralization test (PRNT), the new established DENV-1 specific NS1 antigen capture ELISA was used for detecting the neutralizing abilities of these antibody.</p><p><b>RESULTS</b>Four strains of monoclonal antibodies (mAbs) named 1A1, 1B3, 3D3 and 9D6 and one hyperimmune serum of rabbit were obtained, all of which were approved to have neutralizing abilities to DENV-1 with the PRNT titer of 1:1024, 1:512, 1:256, 1:4096 and 1:4096. MAb 3D3 with the lowest neutralization titer in PRNT had not shown neutralizing ability to DENV-1 in NS1 antigen capture ELISA, while MAbs 1A1, 1B3 and 9D6 and the rabbit hyperimmune serum could protect the C6/36 from being infected by DENV-1 with the neutralization titer of 1:32, 1:32, 1:128 and 1:128 in this assay.</p><p><b>CONCLUSION</b>NS1 antigen capture ELISA could be used to identify antibody neutralizing abilities to DENV, it was a faster and more convenient way to screen antibodies with high neutralization titer and might also be used as one of the methods to evaluate the effects of vaccines.</p>

Clinical evaluation of peripheral vascular disease withwall thickness, stiffness and hemodynamics of femoral artery in type 2 diabetes mellitus / 中国医学影像技术

Objective To evaluate the relationship between femoral artery intima-media thickness (FA-IMT), stiffness, hemodynamics factors of femoral artery and peripheral vascular disease (PVD) in type 2 diabetes mellitus. Methods According to the presence of PVD, 151 patients with type 2 diabetes were divided into PVD group and non-PVD group. FA-IMT was tested with 2D ultrasound. Ds, Dd, PSV, EDV were observed with color flow rate and volume quantitative technology. FA-IMT, FA-stiffness β, peak systolic velocity (PSV) and end diastolic velocity (EDV) were compared between the two groups. Results Compared with non-PVD group, FA-IMT and FA-stiffness β increased in PVD group (P＜0.01), whereas PSV and EDV decreased in PVD group (P＜0.01). There was no relationship between FA-stiffness β and FA-IMT in PVD group (P=0.01). Multiple Logistic regression indicated the presence of PVD symptoms associated closely with FA-stiffness β, PSV, EDV and duration of diabetes. Conclusion Stiffness β plays an important role in the clinical evaluation of type 2 diabetes mellitus.

Evaluation of peripheral vascular disease with femoral artery wall thickness,stiffness and ankle brachium index in type 2 diabetes mellitus / 中华超声影像学杂志

Objective To evaluate the effects of femoral artery wall thickness, stillness and ankle brachium index(ABI) on clinical manifestation of peripheral vascular disease(PVD) in type 2 diabetes mellitus. Methods According to the presence of lower limb and reduced ABI (ABI<0.9), 151 patients with type 2 diabetes were divided into group of patients with and without PVD symptoms, and group of patients with and without reduced ABI. Intima-media thickness of femoral artery (FA-IMT) and stiffness of femoral artery (FA-β) were measured by ultrasound. FA-IMA and FA-β of femoral arteries were compared between group of patients with and without PVD symptoms as well as between group of patients with and without reduced ABI. Correlation between FA-IMT and FA-β was analyzed. Factors affecting symptoms of lower limb and ABI were evaluated by multiple logistic regression analysis. Results FA-IMT and FA-β in group of PVD symptoms were higher than those in group without PVD symptoms. Similarly, patients with reduced ABI had greater FA-IMT and FA-β than those without. However,there was no correlation between FA-IMT and FA-β in group of PVD symptoms. Multiple logistic regression analysis indicated that the presence of PVD symptoms was associated closely with increased FA-β, whereas reduced ABI was associated closely with FA-IMT. Conclusions The stiffening of arterial wall has a significant impact on PVD manifestations,particularly on the leg symptoms in patients with type 2 diabetes.

Expression and identification of H5 subtype hemagglutinin of avian influenza A virus in insect cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To clone and express avian influenza A virus [A/Hong Kong/482/97(H5N1)] H5 subtype hemagglutinin in baculovirus-insect cell expression system and investigate the antigenicity and bioactivity of the recombinant protein.</p><p><b>METHODS</b>H5 gene of influenza A virus was amplified by PCR. The recombinant bacmid was obtained by cloning the gene to the donor plasmid of pFastBacHTB and transformed into DH10Bac competent cells. The recombinant baculovirus stock was prepared by transfecting the recombinant bacmid DNA into the insect cell line for protein expression after amplification. Immunofluorescene assay (IFA) and Western blotting were performed to identify the antigenicity of the recombinant protein, and hemagglutination assay was used to identify its bioactivity.</p><p><b>RESULTS</b>The recombinant his-H5 protein was expressed in the insect cells with a relative molecular mass of 64,000, which showed erythrocyte-agglutinating activities with the red blood cells of guinea pig. Western blotting and IFA demonstrated that the recombinant his-H5 could be recognized and bound by standard anti-H5 sera.</p><p><b>CONCLUSION</b>The recombinant his-H5 with a post-translation modification is successfully obtained in insect cells, which may provide a potential source for further study of the antigen's biological function and for production of the subunit vaccine or monoclonal antibodies.</p>

Antigenicity analysis of nucleocapsid proteins of 3 human coronaviruses SARS-CoV, 229E and OC43 with their monoclonal antibodies / 南方医科大学学报

<p><b>OBJECTIVE</b>To prepare and characterize monoclonal antibodies (mAbs) against the recombinant nucleocapsid (N) protein of 3 human coronaviruses SARS-CoV, 229E and OC43 and study the antigenic relationship between the 3 N proteins.</p><p><b>METHODS</b>BALB/c mice were immunized with the recombinant N proteins of SARS-CoV, 229E and OC43 to obtain the mAbs by means of hybridoma. Screening and identification of the mAbs were performed using indirect enzyme-linked immunosorbent assay (ELISA), Western blotting and indirect immunofluorescence assay. Cross-reactivity between the N proteins of the 3 coronaviruses was analyzed with the prepared mAbs.</p><p><b>RESULTS</b>The mAbs against the recombinant N proteins of SARS-CoV, 229E and OC43 were obtained, which reacted specifically with the corresponding viral N protein as shown by indirect ELISA, Western blotting and indirect immunofluorescence assay. No cross-reactivity was found between the 3 N proteins.</p><p><b>CONCLUSION</b>The prepared mAbs against the recombinant N proteins may provide valuable assistance in studying antigenic relationships of N proteins between the 3 human coronaviruses.</p>

Cloning, purification, and antigenic characterization of three recombinant fragments derived from SARS-CoV S1 domain / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>The present study aimed to clone and express three fragments of genomic RNA derived from SARS associated coronavirus (SARS-CoV) S1 domain and to study its immunogenicity.</p><p><b>METHODS</b>The S1 domain gene was amplified by PCR with specific primers and was inserted into the prokaryotic expression vector pQE-30. Three fragments (40-751, 746-1344 and 746-2001 bp) derived from S1 domain produced after the recombinant plasmid (pQE-30/S1) was digested by restriction endonucleases. The three fragments were cloned into pQE-30 and expressed in M15 strains of Escherichia coli. The expression products, designated S1a, S1b and S1c respectively, were purified by Ni affinity chromatography. The immunogenicity was analyzed by Western Blot and ELISA using serologically confirmed sera from SARS patients and the sera from healthy donors was used as control at the same assay.</p><p><b>RESULTS</b>Three recombinant plasmids (pQE-30/S1a, pQE-30/S1b, pQE-30/S1c) were constructed.Fusion proteins with relative molecular mass of 26,700, 22,500 and 46,000 dalton were successfully expressed with amounts of 35%, 35% and 30% of total cell protein and purified by Ni affinity chromatography, respectively. Western Blot and ELISA analysis showed that the S1c protein could be specifically recognized by the sera from SARS patients.</p><p><b>CONCLUSION</b>The recombinant S1c protein was a good immunogen and has the potential to be used as a vaccine against SARS-CoV infection.</p>