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1.
Neuroscience Bulletin ; (6): 69-80, 2022.
Article in English | WPRIM | ID: wpr-929078

ABSTRACT

Sodium salicylate is an anti-inflammatory medication with a side-effect of tinnitus. Here, we used mouse cochlear cultures to explore the effects of salicylate treatment on cochlear inner hair cells (IHCs). We found that IHCs showed significant damage after exposure to a high concentration of salicylate. Whole-cell patch clamp recordings showed that 1-5 mmol/L salicylate did not affect the exocytosis of IHCs, indicating that IHCs are not involved in tinnitus generation by enhancing their neuronal input. Instead, salicylate induced a larger peak amplitude, a more negative half-activation voltage, and a steeper slope factor of Ca2+ current. Using noise analysis of Ca2+ tail currents and qRT-PCR, we further found that salicylate increased the number of Ca2+ channels along with CaV1.3 expression. All these changes could act synergistically to enhance the Ca2+ influx into IHCs. Inhibition of intracellular Ca2+ overload significantly attenuated IHC death after 10 mmol/L salicylate treatment. These results implicate a cellular mechanism for tinnitus generation in the peripheral auditory system.


Subject(s)
Animals , Mice , Calcium , Exocytosis , Hair Cells, Auditory, Inner , Sodium Salicylate/pharmacology , Tinnitus/chemically induced
2.
Chinese Medical Journal ; (24): 883-888, 2019.
Article in English | WPRIM | ID: wpr-772179

ABSTRACT

BACKGROUND@#Gestational diabetes mellitus (GDM) is usually diagnosed between 24th and 28th gestational week using the 75-g oral glucose tolerance test (OGTT). It is difficult to predict GDM before 24th gestational week because fast plasma glucose (FPG) decreases as the gestational age increases. It is controversial that if FPG ≥5.1 mmol/L before 24th gestational week should be intervened or not. The aim of this study was to evaluate the value of FPG to screen GDM before 24th gestational week in women with different pre-pregnancy body mass index (BMI).@*METHODS@#This was a multi-region retrospective cohort study in China. Women who had a singleton live birth between June 20, 2013 and November 30, 2014, resided in Beijing, Guangzhou and Chengdu, and received prenatal care in 21 selected hospitals, were included in this study. Pre-pregnancy BMI, FPG before the 24th gestational week, and one-step GDM screening with 75 g-OGTT at the 24th to 28th gestational weeks were extracted from medical charts and analyzed. The pregnant women were classified into four groups based on pre-pregnancy BMI: Group A (underweight, BMI < 18.5 kg/m), Group B (normal, BMI 18.5-23.9 kg/m), Group C (overweight, BMI 24.0-27.9 kg/m) and Group D (obesity, BMI ≥28.0 kg/m). The trend of FPG before 24th week of gestation was described, and the sensitivity and specificity of using FPG before the 24th gestational week to diagnose GDM among different pre-pregnancy BMI groups were reported. Differences in the means between groups were evaluated using independent sample t-test and analysis of variance. Pearson Chi-square test was used for categorical variables.@*RESULTS@#The prevalence of GDM was 20.0% (6806/34,087) in the study population. FPG decreased gradually as the gestational age increased in all pre-pregnancy BMI groups until the 19th gestational week. FPG was higher in women with higher pre-pregnancy BMI. FPG before the 24th gestational week and pre-pregnancy BMI could be used to predict GDM. The incidence of GDM in women with FPG ≥5.10 mmol/L in the 19th to 24th gestational weeks and pre-pregnancy overweight or obesity was significantly higher than that in women with FPG ≥5.10 mmol/L and pre-pregnancy BMI <24.0 kg/m (78.5% [62/79] vs. 52.9% [64/121], χ = 13.425, P < 0.001).@*CONCLUSIONS@#FPG decreased gradually as the gestational age increased in all pre-pregnancy BMI groups until the 19th gestational week. Pre-pregnancy overweight or obesity was associated with an increased FPG value before the 24th gestational week. FPG ≥5.10 mmol/L between 19 and 24 gestational weeks should be treated as GDM in women with pre-pregnancy overweight and obesity.


Subject(s)
Adult , Female , Humans , Pregnancy , Blood Glucose , Body Mass Index , Diabetes, Gestational , Blood , Diagnosis , Epidemiology , Fasting , Blood , Gestational Age , Glucose Tolerance Test , Incidence , Prevalence , ROC Curve , Retrospective Studies
3.
China Journal of Orthopaedics and Traumatology ; (12): 1134-1137, 2019.
Article in Chinese | WPRIM | ID: wpr-781677

ABSTRACT

OBJECTIVE@#To investigate the causes of poor incision healing after total knee arthroplasty(TKA) and to propose active preventive and therapeutic measures.@*METHODS@#Total 284 patients undergoing unilateral TKA from September 2016 to June 2018 were enrolled in the study and divided into control group and observation group. Firstly, 150 patients with unilateral TKA from September 2016 to June 2017 were retrospectively analyzed and included in the control group. There were 41 males and 109 females, with an average age of(63.5±7.2) years old. The causes of poor incision healing were discussed about patients themselves and surgical techniques. After analyzing, improvement measures were proposed. Total 134 patients with TKA from July 2017 to June 2018 were included in the observation group. There were 36 males and 98 females, with an average age of(62.4±8.9) years old. The patients in the observation group were treated with improved treatment strategies.@*RESULTS@#Nine patients(6%) had poor incision healing in the control group and 1 patient (0.75%) had poor incision healing in the observation group. The incidence of poor incision healing was significantly different between the two groups (²=5.750, <0.01).@*CONCLUSIONS@#In order to prevent the poor incision healing after TKA, perioperative management and the operation techniques including stable, accurate, rapid and clean skills should be improved, leading to reduce the complications of incision and improve the recovery rate of patients.


Subject(s)
Aged , Female , Humans , Male , Middle Aged , Arthroplasty, Replacement, Knee , Retrospective Studies , Treatment Outcome
4.
Biomedical and Environmental Sciences ; (12): 79-87, 2017.
Article in English | WPRIM | ID: wpr-296512

ABSTRACT

<p><b>OBJECTIVE</b>To explore the association between body mass index (BMI) and all-cause mortality among the elderly in Beijing.</p><p><b>METHODS</b>This analysis was based on the Beijing multidimensional longitudinal study of aging (BLSA), which included 2,090 subjects over 55 years old and was followed-up from 1992 to 2012. BMI-mortality curves were drawn to find the optimal BMI range with the lowest mortality. Cox proportional hazard models were used to obtain the hazard ratios (HRs) for BMI and BMI changes in the overall population and in specific stratified populations.</p><p><b>RESULTS</b>During follow-up, 1,164 deaths were recorded; BMI-mortality curve was U-shaped, with the lowest mortality at a BMI of approximately 25 kg/m2. After adjusting for gender, age, smoking, drinking and some pre-existing diseases, HRs for underweight, overweight and obesity compared with normal weight were 1.372 (95% CI: 1.154-1.631), 0.767 (95% CI: 0.666-0.884) and 0.871 (95% CI: 0.830-1.246), respectively. HR for BMI drop was 3.245 (95% CI: 0.824-12.772) in the underweight group and 1.892 (95% CI: 0.830-1.246) in the normal weight group, HR for BMI rise was 1.795 (95% CI: 1.243-2.591) in normal weight group and 1.962 (95% CI: 1.202-3.203) in the overweight group.</p><p><b>CONCLUSION</b>Keeping BMI in an overweight status and stable is related to a reduced mortality.</p>


Subject(s)
Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Beijing , Body Mass Index , Chronic Disease , Mortality , Cohort Studies , Proportional Hazards Models , Risk Factors
5.
Chinese Journal of Contemporary Pediatrics ; (12): 1143-1148, 2014.
Article in Chinese | WPRIM | ID: wpr-289515

ABSTRACT

<p><b>OBJECTIVE</b>To explore the mechanism and effect of maternal high-fat diet before and during pregnancy on bone growth of neonatal offspring rats.</p><p><b>METHODS</b>Forty female Sprague-Dawley rats were divided into high-fat diet and control groups (n=20) that were fed with 35% high-fat diet and standard chow, respectively. After 8 weeks, 8 female rats from each group were sacrificed for liver pathological examinations and the other female rats were mated with male rats and fed continuously with 35% high-fat diet and standard chow throughout gestation, respectively. The body lengths (from apex nasi to end of tail) of the offspring rats from both groups were measured within 24 hours after birth. Enzyme-linked immunosorbent assay was used to detect serum insulin-like growth factor (IFG-I) levels. Liver pathological changes were observed under a light microscope. The expression of insulin receptor substrate 1 (IRS-1) and phosphorylation IRS-1 (Phospho-IRS-1) in tibia and femur samples were detected by immunohistochemistry. The expression of mitogen-activated protein kinase (MAPK) and phosphorylation MAPK (Phospho-MAPK), phosphatidylinositol 3-kinase (PI3K) and phosphorylation PI3K (Phospho-PI3K), protein kinase B (AKT1) and phosphorylation AKT1 (Phospho-AKT1) in tibia and femur samples were detected by Western blot.</p><p><b>RESULTS</b>The offspring rats from the high-fat diet group showed a significant shorter body length compared with those from the control group (P<0.05). The level of serum IGF-I in offspring rats from the high-fat diet group decreased by 20.1% in comparison to those from the control group, but there was no significant difference between the two groups (P>0.05). Fatty degeneration was found in livers of both high-fat diet-fed maternal rats and their offspring rats under a light microscope. There were no significant differences in IRS-1 and Phospho-IRS-1 expression in chondrocytes of tibia and femur samples between the offspring rats of the two groups (P>0.05). The protein expression of MAPK in chondrocytes of tibia and femur samples of offspring rats from the high-fat diet group was higher than that from the control group (P<0.05). There were no significant differences of PI3K and AKT1/Phospho-AKT1 between the offspring rats of the two groups (P>0.05).</p><p><b>CONCLUSIONS</b>A maternal high-fat diet before and during pregnancy may affect the bone growth of offspring rats in utero, which is possibly associated with the decreased IGF-I level. However, further study on the exact mechanism of IGF-I on the bone growth is needed.</p>


Subject(s)
Animals , Female , Pregnancy , Rats , Bone Development , Diet, High-Fat , Insulin-Like Growth Factor I , Liver , Pathology , MAP Kinase Signaling System , Maternal Nutritional Physiological Phenomena , Phosphatidylinositol 3-Kinases , Physiology , Proto-Oncogene Proteins c-akt , Physiology , Rats, Sprague-Dawley
6.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 1608-1610, 2013.
Article in Chinese | WPRIM | ID: wpr-231636

ABSTRACT

<p><b>OBJECTIVE</b>To study the objective diagnostic mechanisms on Chinese medical (CM) syndrome patterns of rheumatoid arthritis (RA), and to research different titers of rheumatoid factor (RF)/citrullinated protein antibody (CCP) in CM syndrome patterns of RA.</p><p><b>METHODS</b>Totally 230 early RA patients were assigned to five CM syndrome pattern groups, i.e., the dampness-heat blockage group (50 cases), the cold-dampness blockage group (50 cases), the Shen-qi deficiency-cold group (50 cases), the Gan-Shen yin deficiency group (40 cases), and the blood stasis blockage group (40 cases). Another 100 healthy subjects were recruited as the healthy control group. RF-IGM, RF-IGA, RF-IGG, and anti-CCP antibody were detected and compared.</p><p><b>RESULTS</b>The titers of RF-IGM, RF-IGA, RF-IGG, and anti-CCP antibody were higher in all groups than in the healthy control groups (P < 0.01). As for the 5 groups, RF-IGM, RF-IGA,RF-IGG, and anti-CCP antibody were higher in the RA active stage than in the nonactive stage. They were higher in the dampness-heat blockage group in the RA active stage than in the Shen-qi deficiency-cold group, the Gan-shen yin deficiency group, and the blood stasis blockage group.</p><p><b>CONCLUSION</b>Titers of RF-IGM, RF-IGA, RF-IGG, and anti-CCP antibody could be taken as judging indicators for differentiating objective lab indices of CM syndromes and assessing the active stage of RA.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Arthritis, Rheumatoid , Blood , Diagnosis , Allergy and Immunology , Autoantibodies , Blood , Case-Control Studies , Medicine, Chinese Traditional , Peptides, Cyclic , Allergy and Immunology , Rheumatoid Factor , Allergy and Immunology
7.
Chinese Journal of Gastrointestinal Surgery ; (12): 1136-1138, 2012.
Article in Chinese | WPRIM | ID: wpr-312333

ABSTRACT

<p><b>OBJECTIVE</b>To observe the impact of Roux-en-Y gastric bypass on quality of life in non-obese patients with type 2 diabetes mellitus.</p><p><b>METHODS</b>Thirty-seven non-obese patients with type 2 diabetes mellitus who underwent Roux-en-Y gastric bypass were prospectively studied. A 36-item short form healthy survey questionnaire(SF36), the diabetes treatment satisfaction questionnaire(DTSQ), and quality of life scale for patients with type 2 diabetes mellitus(DMQLS) were used to evaluate the quality of life for all the non-obese patients with type 2 diabetes mellitus preoperatively and 12 months postoperatively.</p><p><b>RESULT</b>The blood glucose and lipid indexes were significantly decreased after operation(all P<0.05). SF36 showed the physical and mental synthesis scores at 12 month after operation were 74.6±18.3 and 79.8±14.9 respectively, higher than those at one week before operation(54.9±15.1 and 56.4±17.8, both P<0.01). DTSQ showed treatment satisfaction score was increaced significantly after operation(29.2±7.1 vs. 15.4±5.6, P<0.01). The quality of life evaluated by DMQLS, was also significantly improved(P<0.01).</p><p><b>CONCLUSION</b>Roux-en-Y gastric bypass can significantly improve the quality of life for non-obese patients with type 2 diabetes mellitus.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Diabetes Mellitus, Type 2 , Psychology , General Surgery , Gastric Bypass , Methods , Psychology , Laparoscopy , Methods , Psychology , Obesity , Quality of Life , Retrospective Studies , Surveys and Questionnaires , Treatment Outcome
8.
Chinese Journal of Microsurgery ; (6): 280-282, 2011.
Article in Chinese | WPRIM | ID: wpr-671581

ABSTRACT

ObjectiveTo introduction of perforator flaps,muscle flaps pedicled by descending branch of lateral circumflex femoral artery,method and their clinical application that multiple soft tissue defects of hand are repaire by muliplefoliated tissue flap only branch lateral circumflex femoral artery.MethodsFifteen patients with multiple soft tissue defects of hand were repaired muliplefoliated tissue flap only pedicled bydescending branch lateral circumflex femoral artery.At first,the anterolateral thigh perforator flap was designed and harvested according to the soft tissue defects of hand, then the descending branch lateral circumflex femoral artery was dissected at the same time the segmented perforator flap,fascia lata flap,rectus femoris muscle flap, vastus lateralis muscle flap, vastus intermedius muscle flap and distal spatium intermusculare flap were harvested in need according to distance among soft tissue defects.The muliplefoliated tissue flap was harvested only pedicled by descending branch lateral circumflex femoral artery, at last muscle flaps and fascia lata flaps were covered by skin graft, so the multiple soft tissue defects of hand were repaired in one time.ResultsNo vascular crisis happened. All skin grafts survived well, the contour of all repaired soft tissue defects was good and protective feeling was recovered by skin grafts of all flaps. All cases were got follow-up and the range was from 6 to 20 months(the average was 8.7 months).Wound of donor site healed well, muscle strength of quadriceps and motion of knee were normal. Three cases were excellent,nine cases were well and 3 cases were good, according to upper extremity function evaluation criteria of Chinese Medical Society for the Surgery of the Hand, the rate of good was 80 percent.ConclusionMultiple soft tissue defects of hand can be repaired by muliplefoliated flap only pedicled by descending branch of lateral circumflex femoral artery. Its advantages included reduction of operation time and treatment, good recovery of hand contour and function. It is a good method to repair multiple soft tissue defects of hand.

9.
World Journal of Emergency Medicine ; (4): 118-121, 2010.
Article in Chinese | WPRIM | ID: wpr-789473

ABSTRACT

BACKGROUND:Stroke is the leading cause of death and long-term disability. This study was undertaken to investigate the factors influencing daily activities of patients with cerebral infarction so as to take interventional measures earlier to improve their daily activities. METHODS:A total of 149 patients with first-episode cerebral infarction were recruited into this prospective study. They were admitted to the Encephalopathy Center, Department of Neurology, the First Affiliated Hospital of Wenzhou Medical College in Zhejiang Province from August 2008 to December 2008. The baseline characteristics of the patients and cerebral infarction risk factors on the first day of admission were recorded. White blood cell (WBC) count, plasma glucose (PG), and many others of laboratory targets were col ected in the next morning. Barthel index (BI) was calculated at 2 weeks and 3 months respectively after onset of the disease at the outpatient clinic or by telephone cal . Lung infection, urinary tract infection and atrial fibrillation if any were recorded on admission. The National Institute of Health Stroke Scale (NIHSS) scores and the GCS scores were recorded within 24 hours on and after admission, at the second week, and at the third month after the onset of cerebral infarction respectively. RESULTS:The factors of BI at 2 weeks and 3 months after onset were the initial PG level, WBC count and initial NIHSS scores. Besides, urinary tract infection on admission was also the factor for BI at 3 months. CONCLUSION:Active measures should be taken to control these factors to improve the daily activities of patients with cerebral infarction.

10.
Chinese Journal of Biotechnology ; (12): 794-800, 2007.
Article in Chinese | WPRIM | ID: wpr-327945

ABSTRACT

RNA interference (RNAi) is a powerful tool in gene function research. In order to investigate the role of GP2, GP3 and GP4 of porcine reproductive and respiratory syndrome virus (PRRSV) in the viral replication, small interference RNAs (siRNAs) directed to ORF2, ORF3 and ORF4 were designed and 12 short hairpin RNA (shRNA) expression vectors were constructed (designed as 21,22,23,24,31,32,33,34,41,42,43 and 44). Cells treated with shRNA expression vectors were infected by PRRSV. The effective shRNA expression vectors were selected by fluorescent quantatitive PCR (FQ-PCR). The virus titer of supernatant of the cells treated with effective shRNA expression vectors (23,24,31,34 and 41) were reduced by 184 to 4.65 folds compared with that of controls.


Subject(s)
Genetic Vectors , Open Reading Frames , Porcine respiratory and reproductive syndrome virus , Genetics , RNA Interference , RNA, Small Interfering , Genetics , Metabolism , RNA, Viral , Chemistry , Metabolism , Transduction, Genetic , Viral Proteins , Genetics , Virus Replication , Genetics
11.
Chinese Journal of Biotechnology ; (12): 701-706, 2006.
Article in Chinese | WPRIM | ID: wpr-286223

ABSTRACT

To identify the epitope of SARS-CoV spike protein specific neutralizing monoclonal antibody (MAb) 2C5. The antibody was used as target and three rounds of bio-panning were conducted with phage-display peptide library. After the third panning, 20 phage-plague clones were randomly picked and analyzed for the binding ability with the MAb 2C5 by ELISA. The display sequence analysis demonstrated that among the twenty phage clones, eight clones displayed the same seven-peptide TPEQQFT. All these eight phage-clones showed strongest binding activity with 2C5 in phage ELISA analysis. Furthermore, phages displaying peptide TPEQQFT could specifically inhibit the binding of MAb 2C5 with SARS-CoV spike protein. The results demonstrated that TPEQQFT is a mimic epitope peptide containing neutralizing MAb 2C5. This study may provide information for further structural and functional analysis of spike protein and development vaccine for severe acute respiratory syndrome.


Subject(s)
Amino Acid Sequence , Antibodies, Monoclonal , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , Epitopes , Membrane Glycoproteins , Chemistry , Allergy and Immunology , Molecular Sequence Data , Peptide Library , Severe acute respiratory syndrome-related coronavirus , Allergy and Immunology , Spike Glycoprotein, Coronavirus , Viral Envelope Proteins , Chemistry , Allergy and Immunology
12.
Chinese Journal of Biotechnology ; (12): 931-939, 2006.
Article in Chinese | WPRIM | ID: wpr-325447

ABSTRACT

The Fusion (F) and Haemagglutinin-Neuraminidase (HN) genes of Newcastle disease virus (NDV) and the glycoprotein B (gB) gene of infectious laryngothracheitis virus (ILTV) as well as a LacZ reporter gene were all inserted into a nonessential gene of fowlpox virus (FPV) 017 strain by homologous recombination. The NDV and ILTV genes were each under the control of a fowlpox virus immediate early/late promoter (LP2EP2) while the LacZ reporter gene expression cassette was regulated by a P11 late promoter. A recombinant FPV harboring the F, HN and gB genes as well as the LacZ gene, designated as rFPV-F/HN/gB/LacZ, was obtained after ten cycles of blue plaque purification. The presence of the NDV and ILTV genes was confirmed by PCR. The expression of the recombinant proteins in rFPV-F/HN/gB/LacZ were characterized by Western blot (F and gB proteins) and indirect immunofluorescence test (F, HN and gB proteins). The results demonstrated that all four foreign proteins, which were encoded within a 10 kb gene fragment, could be expressed authentically and efficiently. Compared to the parental virus, rFPV-F/HN/gB/LacZ showed no obvious difference with respect to virus replication and cytopathogenic effects in chicken embryo fibroblasts (CEF) cell culture. Overall, our work suggests that FPV can be a useful live virus vector for the expression of multi- foreign genes against multiple avian pathogens.


Subject(s)
Animals , Cloning, Molecular , Fibroblasts , Virology , Fowlpox virus , Genetics , Gene Expression , Genetic Engineering , Methods , HN Protein , Genetics , Herpesvirus 1, Gallid , Genetics , Physiology , Newcastle disease virus , Genetics , Physiology , Plasmids , Genetics , Transfection , Viral Envelope Proteins , Genetics , Viral Fusion Proteins , Genetics
13.
Chinese Journal of Biotechnology ; (12): 211-215, 2005.
Article in Chinese | WPRIM | ID: wpr-249923

ABSTRACT

Based on the genomic sequence of SARS-CoV strain BJ101, antigenic immunodominant genes coding for the structure proteins of SARS-CoV were predicted by bio-informatics methods, and two chimeric genes A and B with multi-immunodominants lined up by Gly-Pro-Gly linker were synthesized. The chimeric genes were cloned into plasmid pGEX-6p-1 and expressed in E. coli with IPGT inducing. BALB/c mice were immunized with the purified recombinant fusion protein. The specificity of monoclonal antibodies were tested with a commercial ELISA kit for detecting antibody against SARS-CoV. The results showed that two peptides with molecular weights of 34kD and 35kD expressed by the two chimeric genes could be recognized by SARS patient convalescent serum in Western blot. Six positive hybridoma cell lines stably secreting monoclonal antibodies were selected. The subtype of monoclonal antibody D3C5 is IgG2a, and subtypes of all other five monoclonal antibodies are IgG1. Light chains of all monoclonal antibodies are kappa. With a commercial SARS-CoV antibodies detection ELISA kit, five out of six monoclonal antibodies were positively recognized. In western blot analysis with inactived virus cultures, D3D1 specifically recognized a band of about 180 kD. To further analyse the epitopes corresponding to the monoclonal antibodies, six oligoes (S1-S6) from S gene were synthesized and expressed. The results showed that the monoclonal antibodies D3D1 and D3C5 specifically recognized expression product of S2 and S5 oligoes, respectively. The S2 and S5 oligoes are corresponding to 447-458aa and 789-799aa of SARS-CoV S protein respectively.


Subject(s)
Animals , Female , Humans , Mice , Antibodies, Monoclonal , Allergy and Immunology , Antibodies, Viral , Allergy and Immunology , Antibody Specificity , Epitopes , Genetics , Escherichia coli , Genetics , Metabolism , Hybridomas , Bodily Secretions , Membrane Glycoproteins , Allergy and Immunology , Mice, Inbred BALB C , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Severe acute respiratory syndrome-related coronavirus , Allergy and Immunology , Spike Glycoprotein, Coronavirus , Viral Envelope Proteins , Allergy and Immunology
14.
Chinese Journal of Biotechnology ; (12): 526-531, 2004.
Article in Chinese | WPRIM | ID: wpr-270093

ABSTRACT

With the application of gE gene deleted pseudarabies virus (PRV) vaccine worldwide, a corresponding differential diagnosis based on gE glycoprotein is needed in the project of PRV eradication. In this study, PRV gE gene without signal and transmembrane region was amplified by PCR and cloned into pGEX-6P-1, generated pGEX-gE. After transformation of BL21 with pGEX-gE, an expressed fusion protein(about 63kD) was identified by SDS-PAGE. The recombinant proteins are produced as inclusion bodies. By changing the inductive conditions, the formation of inclusion bodies was inhibited and tended to increase the percentage of soluble recombinant protein. The antigenic reactivity of the recombinant protein was confirmed by Western blotting with polyclonal antibodies against PRV. Using purified recombinant tgE as antigen, an ELISA was developed to detect sera of PRV infected pigs and sera of pigs immunized with gE-deleted PRV vaccine. The total of 400 serum samples collected from field were comparatively tested with the tgE-ELISA and a commercial competitive ELISA based on monoclonal antibody against gE, the results indicated that the coincidental rate between the two tests is about 94%.


Subject(s)
Animals , Diagnosis, Differential , Enzyme-Linked Immunosorbent Assay , Herpesvirus 1, Suid , Allergy and Immunology , Pseudorabies , Diagnosis , Pseudorabies Vaccines , Allergy and Immunology , Recombinant Proteins , Allergy and Immunology , Swine , Vaccination , Viral Envelope Proteins , Genetics , Allergy and Immunology
15.
Chinese Journal of Pediatrics ; (12): 589-594, 2004.
Article in Chinese | WPRIM | ID: wpr-340252

ABSTRACT

<p><b>OBJECTIVE</b>Alport syndrome (AS) is a progressive hereditary nephritis presented with hematuria and renal failure, frequently associated with sensorineural deafness and ocular lesions. So far, more than 300 gene mutations in AS have been identified which provides a better way to analyze the association between genotype and phenotype. It is hard to understand all the phenotype according to the gene mutations, because the structure and function changes of the relevant protein, alpha5(IV) chain, encoded by mutated COL4A5 gene are rare to know. This study aimed to detect the proteins structure encoded by COL4A5 gene with different missense mutations and to analyze the effect of gene mutations on the secondary structure of alpha5(IV) chain and structure-phenotype relations.</p><p><b>METHODS</b>Two X-linked AS patients with different missense mutations (g.3246G > T resulting in p.G1015V and g.3290G > A resulting in p.G1030S, respectively) diagnosed by clinical manifestations, family history and skin or renal biopsy examinations, as well as a control were included in this study. The fragments of cDNA with the two mutations, respectively, and that of corresponding cDNA from the control were expressed in E. coli. The secondary structure of the recombinant polypeptides were analyzed by using circular dichroism (CD) spectroscopy.</p><p><b>RESULTS</b>CD spectra of the control exhibited a negative peak near 200 nm whereas that of the patient 1 exhibited a negative peak near 220 nm. Furthermore, the magnitude of the negative peak of patient 1 decreased from -9000 deg x cm2 x dmol(-1) to -4000 deg x cm2 x dmol(-1) as compared with that of the control. CD spectra of the patient 2 were slightly changed with the negative peak remaining near 220 nm but the magnitude increasing from -9000 deg x cm2 x dmol(-1) to -11000 deg x cm2 x dmol(-1) as compared with that of the control. In addition, the secondary structure of the control polypeptide was mainly composed of beta-sheet and random coil without alpha-helix, whereas that of the patient 1 presented 12.9% alpha-helix. Although the secondary structure of polypeptide of the patient 2 was also mainly composed of beta-sheet and random coil, the composition of beta-sheet reduced and random coil increased.</p><p><b>CONCLUSION</b>Although the glycine substitutions existed in the same domain of alpha5(IV) chain, the patient 1 with the severe AS phenotype and g.3246G > T mutation, and patient 2 with the mild AS phenotype and g.3290G > A mutation were revealed with different secondary structures of alpha5(IV) chain. Moreover, the secondary structure changes of alpha5(IV) chain were consistent with their corresponding phenotype severity.</p>


Subject(s)
Humans , Collagen Type IV , Genetics , Mutation, Missense , Nephritis, Hereditary , Genetics , Phenotype , Point Mutation , Protein Structure, Secondary , Genetics , Spectrum Analysis
16.
Chinese Journal of Biotechnology ; (12): 623-627, 2003.
Article in Chinese | WPRIM | ID: wpr-259136

ABSTRACT

Multiple epitopes from one or more viruses can be lined up and co-expressed in one vector to generate multi-epitopes DNA vaccines. In the study, four recombinant plasmids were constructed based on HA and NP gene of avian influenza virus (AIV) (H5N1): (1) pIRES/HA, carrying the complete HA gene; (2) pIRES/tHA, carrying a truncated HA gene fragment of major neutralizing antigenic epitopes; (3) pIRES/tHA-NPep, in which three CTL epitopes of NP gene of AIV were fused to the truncated HA from the C-terminal; and (4) pIRES/tHA-NPep-IFN-gamma, which was constructed by replacing neo gene in pIRES/ tHA-NPep with IFN-y of chicken. Fifty five SPF chickens were randomly divided into five groups and immunized with the above four constructs and control plasmid. Each chicken was intramuscally immunized with 200 microg plasmid DNA three times in a two-week interval. Two weeks after the third immunization, chickens were injected with H5N1 subtype avian influenza virus. Before the virus loading no detectable antibodies to HA were found in the chicken serum; but high levels of HI antibodies were detected in the serum of the survived chickens. The percentages of CD4+ and CD8+ T lymphocyte in peripheral blood of immunized chickens increased steadily after the vaccination. After virus loading all chickens in the control group died within three to eight days, and the survival rates of the four DNA vaccine groups were as follows: pIRES/HA, 54.5%; pIRES/tHA, 30%, pIRES/ tHA-NPep, 36.3%, pIRES/tHA-NPep-IFN-gamma, 50%. These results indicated that multi-epitopes DNA immunization can induce immune response and protect chickens from homologous virus loading.


Subject(s)
Animals , Chickens , Epitopes , Allergy and Immunology , Influenza A Virus, H5N1 Subtype , Allergy and Immunology , Virulence , Influenza in Birds , Allergy and Immunology , Virology , Vaccines, DNA , Allergy and Immunology
17.
Chinese Journal of Obstetrics and Gynecology ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-683011

ABSTRACT

0.05).The required dose of LT4 during the second and third trimester was respectively,remarkably increased compared to pre-gestational period (P

18.
Journal of Applied Clinical Pediatrics ; (24)1994.
Article in Chinese | WPRIM | ID: wpr-639869

ABSTRACT

Objective To explore the clinical significance of mycoplasma pneumoniae(MP) antibody IgM positive with high titers.Methods Two hundred and sixty-seven cases of respiratory tract infections of MP with MP-IgM positive were confirmed in our hospital during Jun.2005 to Aug.2008,of which 63 cases with significantly increased titers of MP-IgM antibodies(MP-IgM≥1:1 280).A prospective observation and later analysis of the clinical manifestations of the 63 cases were carried out.Results 1.Among 63 cases,6 cases were diagnosed as upper respiratory tract infection,12 cases as acute bronchitis and 45 cases as pneumonia,respectively.2.The 63 cases pre-sented mainly with fever and cough,of them 59 cases had fever and 45 cases with paroxysmal cough,39 cases could be found crackles on physical examination.3.Extrapulmonary manifestations could be found in many cases,including heart complications in 21 cases,abnormal liver functions in 35 cases,gastrointestinal manifestations in 22 cases,neurological manifestations in 1 case,dermatological disorders in 7 cases and renal manifestations in 3 cases.4.Patterns of chest X-ray findings of the 63 cases with MP infections were varied,of which 45 cases showed pulmonary abnormality.Chest X-ray findings of the 45 children included 12 cases with interstitial infiltration,6 cases with lobar condensation,15 cases with patch shadows,4 cases with enlarged porta pulmonis and 8 cases with reticulonodular infiltrates.Three cases were complicated with pleural effusion and 2 children with segmental atelectasis.5.The cases were given azithromycin,10 mg/(kg?d),a daily dose for 5 days and intermitted for 4 days.All cases repeated the treatment courses for 1 to 4 times.The mean duration of fever was 6.6 days,cough could last 11.7 days and the crackles on physical examination could not be found 7.5 days later after treatment.The abnormality of chest X-ray findings dissolved in 38 cases 2-3 weeks later after treatment.Conclusions Cases of MP infections with high titers of MP-IgM positive had severe symptoms,of which more than 90% patients presenting with lower respiratory tract infections,mainly with pneumonia.Recovery of chest radiological finding was posterior to clinical improvement,so timely and regular chest X-ray examinations were essential after treatment and could avoid the false ceasing of treatment of the cases.Patients of MP infections with high titers of IgM antibodies positive have good prognosis.

19.
Journal of Applied Clinical Pediatrics ; (24)1986.
Article in Chinese | WPRIM | ID: wpr-640198

ABSTRACT

Objective To analyze the trend of disease constitution of hospitalized neonates in China-Japan Friendship Hospital and provide a scientific basis for the focus adjustment of perinatal health.Methods Six hundred and ninety-eight neonates were admitted during Jan.2004 to Dec.2008.Relevant data of all cases were retrospectively collected.The discharge diagnosis of the neonates was counted accor-ding to the year,and the trends of the neonatal disease constitution of the 5 years were analyzed.Results 1.The first 10 diseases of hospita-lized neonates were centralized,and the first 3 diseases including premature infants,neonatal hypoxic-ischemic encephalopathy(HIE) and neonatal hyperbilirubinemia accounted for 42.84% of the neonate diseases.2.The characteristics of neonatal disease constitution were as fo-llows:the disease constitution of premature infants always located within the first 3 diseases;the location of neonatal HIE went ahead from number 8 to number 1 or 2,and its constitution proportion increased from 4.93% to more than 15.00%(P

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