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1.
China Pharmacy ; (12): 1961-1966, 2020.
Article in Chinese | WPRIM | ID: wpr-825009

ABSTRACT

OBJECTIVE:To c ompare the difference of volatile oil and fatty oil constituents from Cinnamomum migao in different sources. METHODS :The steam distillation method and Soxhlet extraction mothod were used to extract volatile oil and fatty oil from C. migao in different sources respectively ,and the extraction rates were calculated ;GC-MS was used to analyze volatile oils and fatty oils constituents from C. migao in different sources. The compounds were searched and matched through NIST 17,WILEY 275 databases and mass spectrometry computer date system. The relative percentage content of each constituent was calculated by peak area normalization method. RESULTS :The extraction rates of the volatile oils from 4 batches of C. migao in different sources were 3.1%,4.5%,6.2% and 5.5%,respectively;the extraction rates of the fatty oils from C. migao were 6.2%,8.3%,10.5% and 9.4%,respectively. A total of 87 constituents were identified in 4 batches of volatile oils of C. migao in different sources ,of which 104 constituents were separated from S 1,67 were identified ,and the relative percentage content was 90.172%;102 constituents were separated from S2,73 were identified ,and the relative percentage contentwas 88.836%;77 constituents were separated from S 3,57 were identified , with a relative percentage content of 93.972%;87 constituents were separated from S 4,60 were m identified,with a relative percentage content of 95.247% . Among above 87 constituents,48 were monotyloids and their derivat ives,33 were sesquiterpenoids and their derivatives ,4 were aliphatic and 2 were ketones. There were 44 common constituents from the volatile oil of C. migao in different sources ,all of which were terpenoids. The relative percentage content of S 1-S4 were 38.556%,66.776%,88.886% and 90.115%,respectively. Among 44 common constituents ,the relative percentage content of which were all greater than 1% were 1,8-cineole(S2: 6.518%;S4:3.850%;S3:1.655%;S1:1.475%;),4-terpineol(S2:1.591%;S4:1.384%;S3:1.193%;S1:1.182%), α-terpinenol(S3:8.662%;S4:7.173%;S2:6.503%;S1:4.839 %),δ-cadinene(S3:8.597%;S4:5.329%;S2:2.677%; S1:2.547%),elemol(S3:4.781%;S2:4.113%;S1:2.568%;S4:1.897%)and γ-eudesmol(S2:4.061%;S3:2.167%;S1: 1.575%;S4:1.197%). A total of 37 constituents were identified in the 4 batches of fatty oil of the C. migao in different sources , of which 87 constituents were separated from S 1,34 were identified ,and the relative percentage content was 91.072%;69 constituents were separated from S 2,28 were identified ,and the relative percentage content was 90.527%;63 constituents were separated from S 3,23 were identified ,the relative percentage content was 85.297%;71 constituents were separated from S 4,24 were identified ,with relative percentage content of 91.527%. Among above 37 constituents,there were 21 monoterpenes and their derivatives,2 sesquiterpenes,13 aliphatics,and 1 alkane. There were 20 common constituents in fatty oil from C. migao of different sources ,and the relative percentage content in S 1-S4 were 89.667%,89.595%,84.651% and 90.972%,respectively. Among 20 common constituents ,the constituents with relative percentage content greater than 1% were methyl caprate (S4: 59.498%;S1:58.733%;S2:57.552%;S3:26.423%)and methyl dodecanoate (S3:31.434%;S2:26.990%;S1:25.095%; S4:24.334%). CONCLUSIONS :There are differences in volatile oil and fatty oil constituents of C. migao from different sources , and the contents of the same constituent were also different.

2.
Acta Academiae Medicinae Sinicae ; (6): 334-337, 2018.
Article in Chinese | WPRIM | ID: wpr-690334

ABSTRACT

Objective To explore the association between the severity of scrub typhus (ST) and the serum procalcitonin (PCT) level. Methods The clinical data of 58 ST patients who were treated in the First Affiliated Hospital of Xiamen University and confirmed by Xiamen Municipal Center for Disease Prevention and Control (CDC) from January 2016 to October 2017,were retrospectively analyzed. According to clinical manifestations and related laboratory tests,these patients were divided into four grades:Ⅰ,Ⅱ,Ⅲ,and Ⅳ. These four grade groups were compared in terms of age,interval from symptom onset to hospital presentation,hospitalization days,and serum PCT level. Results These 58 patients were divided into four grade groups:grade Ⅰ group (n=17),grade Ⅱ group (n=17),grade Ⅲ group (n=11),and grade Ⅳ group (n=13). No significant difference was found among these four groups in terms of age (F=0.618,P=0.606),interval from symptom onset to hospital presentation (F=1.744,P=0.169),and hospitalization days (F=0.398,P=0.755).However,the median serum PCT level in the grade Ⅳ patients[2.60(1.33,61.08)ng/ml] was significantly higher than those in grade I[0.24(0.10,0.33)ng/ml;Z=-4.63,P=0.000], grade Ⅱ[0.29(0.21,0.51)ng/ml;Z=-4.63,P=0.000], and grade Ⅲ[1.33(0.89,2.41)ng/ml;Z=-2.09,P=0.040].The median serum PCT level in the grade Ⅲ patients was also significantly higher than grade Ⅰ (Z=-4.16,P=0.000)and grade Ⅱ(Z=-3.83,P=0.000).There was no significant difference between the grade Ⅰ patients and grade Ⅱ patients(Z=-1.37,P=0.170).There was significantly positive correlation between PCT level and the severity of ST (r=0.804,P=0.000).Conclusion There is positive correlation between serum PCT level and the severity of ST,and serum PCT level may be a biomarker in assessing the severity of ST.

3.
Chinese Journal of Health Policy ; (12): 40-46, 2018.
Article in Chinese | WPRIM | ID: wpr-703598

ABSTRACT

This paper systematically reviews the latest and relevant literatures and policy documents on the in-tegrated health services in Canada in recent years. Therefore, it summarizes the practice and mode of integrated serv-ice delivery in Ontario, Alberta and Quebec wherein the integration among health organization, health service team, and a series of health services are included. The contributing factors and impeding factors ( the barriers) of organiza-tional integration and specific integration strategy were summarized. Finally, according to the actual practical situa-tion, it is proposed that China should adhere to the government-led approach in promoting the integration of health services, and give a full play to the positive role of the market mechanism. Through strengthening the network man-agement and group service of primary health services, emphasis will be put on health services of population groups and specific diseases. Therefore, integration will be regarded as a strategic priority, increasing incentives and boos-ting promotion of nursing personnel on the process of Integrated Service Delivery, building the health information sys-tem that is conducive to integration in order to continuously advance Hierarchical Diagnosis and bridge the fragmented service system. This will help in providing residents with personalized, convenient, comprehensive, and continuous health services.

4.
China Journal of Orthopaedics and Traumatology ; (12): 557-560, 2017.
Article in Chinese | WPRIM | ID: wpr-324618

ABSTRACT

<p><b>OBJECTIVE</b>To investigate surgical method and clinical curative effects of medial patellofemoral ligament (MPFL) reconstruction with great adductor muscular tendon in treating teenagers' recurrent patellar dislocation.</p><p><b>METHODS</b>From May 2012 to September 2014, 19 patients with recurrent dislocation of patellar, including 6 males and 13 females with an average of 16 years old (ranged from 13 to 17 years), the courses of disease ranged from 3 to 18 months(averaged 6 months). All patients were underwent great adductor muscular tendon transposition to reconstruct medial patellofemoral ligament. The curative effects were evaluated by preoperative and postoperative with Lysholm scores and Patellofemoral angle and Q angle.</p><p><b>RESULTS</b>All patients were followed up from 12 to 18 months with an average of 16.5 months. Primary healing was achieved at stage I. No pain, swelling and patellar dislocation or subluxation occurred. Patellofemoral angle increased from preoperative (-3.8±4.9)° to (10.3±4.1)° postoperatively. Q angle decreased from preoperative(16.4±3.1)° to(10.5±1.2)° postoperatively; Lysholm scores were improved from preoperative (68.6±8.5) to (93.7±6.5) final follow-up (<0.01), and 15 cases got excellent results, 3 good, and 1 fair.</p><p><b>CONCLUSIONS</b>Reconstruction of medial patellofemoral ligament with transposition of great adductor muscular tendon could obviously recover stability of patellar, and it is one of the effective methods for the treatment of teenagers' recurrent patellar dislocation.</p>

5.
Chinese Journal of Infection Control ; (4): 422-425, 2016.
Article in Chinese | WPRIM | ID: wpr-494091

ABSTRACT

Objective To evaluate the effect of multiple intervention measures on perioperative antimicrobial use in pa-tients undergoing typeⅠincision operation in a hospital,and provide basis for rational use of antimicrobial agents. Methods 9 823 patients with type I incision operation in April and October of 2005-2012 were surveyed retrospectively, data of 2005 was as baseline,from 2006 to 2012,multiple measures,including training,examination,supervision,feed-back,and cooperation of relevant departments were conducted,antimicrobial use before and after intervention was com-pared.Results The qualified rate of perioperative antimicrobial use in patients undergoing typeⅠ incision operation in-creased from 14.20% in 2006 to 92.30% in 2012;the rate of combined use of antimicrobial agents was relatively higher (7.00%-9.00%)in 2006-2009,had a downward trend in 2010- 2012,and decreased to 3.20% in 2012.Types of an-timicrobial agents for prophylactic use in typeⅠincision operation in 2006 and 2007 were similar to that of 2005,the main used antimicrobial agents were cephalosporins,penicillin and it’s compounds,and aminoglycosides;the major antimicrobial prophylaxis in 2008-2012 were the first and second generation cephalosporins,penicillin and it’s compounds.Multivariate non-conditional logistic regression analysis showed that age (40 - 59 years old),departments (orthopedics,general surgery,and ophthalmology),and years (2011 and 2012)were the main influencing factors for the qualified use of antimicrobial agents(all P<0.05).Conclusion Implementation of multiple intervention measures can improve the qualified rate of perioperative antimicrobial use in typeⅠ incision operation,reduce types of antimicrobial use and rate of combined antimicrobial use.

6.
Journal of Chinese Physician ; (12): 562-564,570, 2016.
Article in Chinese | WPRIM | ID: wpr-604143

ABSTRACT

Objective To investigate the correlation between carotid artery plaque and blood pressure variability (BPV) in elderly patients with hypertension.Methods One hundred sixty elderly patients with hypertension were divided into plaque and non-plaque groups according to the results of carotid artery ultrasonography.All the patients were measured by ambulatory blood pressure monitoring.The mean blood pressure,mean pulse pressure,and blood pressure variability coefficient of two groups were calculated and compared during whole day,daytime,and nighttime.The related factors of carotid artery plaque were analyzed by multivarite logistic regression analysis.Results The 24 h systolic blood pressure standard deviation,daytime and nighttime of systolic blood pressure standard deviation,daytime diastolic blood pressure standard deviation of plaque group were higher than those of non-plaque group (P < 0.05).The 24 h systolic blood pressure variation (24 h SBPV) and night systolic blood pressure variation (nSBPV) were higher than those of non-plaque group (P <0.01).Multivariate regression analysis results showed that carotic artery plaque was associated positively with 24 h BPV and blood pressure variability coefficient of nighttime (P < 0.05).Conclusions The elderly hypertensive patients with carotid artery plaque is associated positively with 24 h systolic blood pressure variability coefficient and blood pressure variability of nighttime.

7.
Biomedical and Environmental Sciences ; (12): 742-753, 2016.
Article in English | WPRIM | ID: wpr-296545

ABSTRACT

<p><b>OBJECTIVE</b>To trace the source of human H7N9 cases in Huai'an and elucidate the genetic characterization of Huai'an strains associated with both humans and birds in live poultry market.</p><p><b>METHODS</b>An enhanced surveillance was implemented when the first human H7N9 case was confirmed in Huai'an. Clinical specimens, cloacal swabs, and fecal samples were collected and screened by real-time reverse transcription-polymerase chain reaction (RT-PCR) for H7N9 virus. The positive samples were subjected to further RT-PCR and genome sequencing. The phylodynamic patterns of H7N9 virus within and separated from Huai'an and evolutionary dynamics of the virus were analyzed.</p><p><b>RESULTS</b>Six patients with H7N9 infection were previously exposed to live poultry market and presented symptoms such as fever (>38.0 °C) and headaches. Results of this study support the hypothesis that live poultry markets were the source of human H7N9 exposure. Phylogenetic analysis revealed that all novel H7N9 viruses, including Huai'an strains, could be classified into two distinct clades, A and B. Additionally, the diversified H7N9 virus circulated in live poultry markets in Huai'an. Interestingly, the common ancestors of the Huai'an H7N9 virus existed in January 2012. The mean nucleotide substitution rates for each gene segment of the H7N9 virus were (3.09-7.26)×10-3 substitutions/site per year (95% HPD: 1.72×10-3 to 1.16×10-2).</p><p><b>CONCLUSION</b>Overall, the source of exposure of human H7N9 cases in Huai'an was live poultry market, and our study highlights the presence of divergent genetic lineage of H7N9 virus in both humans and poultry specimens in Huai'an.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Animals , Female , Humans , Male , Middle Aged , China , Epidemiology , Evolution, Molecular , Influenza A Virus, H7N9 Subtype , Classification , Genetics , Influenza in Birds , Epidemiology , Virology , Influenza, Human , Epidemiology , Virology , Molecular Epidemiology , Phylogeny , Poultry , Prevalence
8.
Journal of Southern Medical University ; (12): 522-525, 2015.
Article in Chinese | WPRIM | ID: wpr-355336

ABSTRACT

<p><b>OBJECTIVE</b>To prepare rabbit anti-mouse zona pellucida 2 (mZP2) polyclonal antibodies and test their immunoactivity.</p><p><b>METHODS</b>Recombinant proteins of mZP2 expressed in Rosetta transformant was separated by SDS-PAGE, and the gel strips containing the recombinant mZP2 were cut out and emulsified to immunize New Zealand white rabbits. The antibody response of the antiserum was detected by ELISA, and the specificity of the antiserum was verified by immunohistochemical assay. The effect of the antiserum on the binding of oocytes with acrosomal reacted sperm was tested by sperm-egg binding assay.</p><p><b>RESULTS</b>ELISA results showed that the immunized rabbit produced anti-mZP2 antiserum. The antiserum reacted specifically with the zona pellucida of mouse ovarian sections. Sperm-egg binding assay showed that treatment of the oocytes with the anti-mZP2 antiserum caused decreased binding of zona pellucida with the acrosomal reacted sperm by 43.7%.</p><p><b>CONCLUSION</b>We obtained rabbit anti-mouse ZP2 polyclonal antibodies that can inhibit the binding of oocytes with acrosomal reacted sperm.</p>


Subject(s)
Animals , Female , Male , Mice , Rabbits , Antibodies , Allergy and Immunology , Antibody Formation , Egg Proteins , Allergy and Immunology , Immune Sera , Membrane Glycoproteins , Allergy and Immunology , Oocytes , Receptors, Cell Surface , Allergy and Immunology , Recombinant Proteins , Allergy and Immunology , Sperm-Ovum Interactions , Spermatozoa , Zona Pellucida Glycoproteins
9.
Chinese Journal of Behavioral Medicine and Brain Science ; (12): 458-460, 2015.
Article in Chinese | WPRIM | ID: wpr-469421

ABSTRACT

Objective To explore the effects of smoking on serum paraoxonase1 (PON1) activity and carotid atherosclerosis.Methods 478 subjects from residents of health screening for cardiovascular disease were enrolled from June 2012 to July 2014 in Huangpu district,shanghai.Smoking,drinking,exercise and cardiovascular disease risk factor data were recorded and gathered.All subjects accepted carotid artery ultrasound examination and were measured serum PON1 activity.The lowest quartile of serum PON1 activity level was taken as low PON1 activity level.Results (1) Serum PON1 activity in smokers was lower than that in non-smokers ((206.5±25.6) kU/L vs (230.9±38.1)kU/L,P<0.01),incidence of carotid artery atherosclerosis in smoking group was higher than non-smoking group(75.7% vs 56.1%,P<0.01).(2) Multivariate logistic regression analyses indicated smoking,lack of exercise,creatinine,LDL-C were the independence factors of PON1 activity.(3) Multivariate logistic regression analyses indicated smoking,serum PON1 activity,age,gender,systolic pressure,were the independence factors of carotid atherosclerosis.Conclusion Smoking reduces serum activity.Smoking and lower serum PON1 activity level are independent risk factors of carotid atherosclerosis.

10.
International Eye Science ; (12): 1127-1129, 2014.
Article in Chinese | WPRIM | ID: wpr-641851

ABSTRACT

AIM: To investigate the clinical effect for treating bullous keratopathy ( BK ) by anterior corneal stromal acupuncture combined with amniotic membrane transplantation. METHODS: Totally 35 patients ( 35 eyes ) with bullous keratopathy were treated by corneal stromal acupuncture combined with amniotic membrane transplantation. All patients preoperative and postoperative underwent anterior segment OCT, corneal topography, corneal sensitivity and confocal microscopy. To observe postoperative ocular symptoms in patients with BK, recurrence of bulla, changes in corneal thickness, corneal sensitivity and changes in the organizational structure of the layers of the cornea. RESULTS: Thirty-five were followed up for 6-18mo. The symptom of pain disappeared in 32 cases ( 91%) in the first day after operation and did not recur during follow-up. The symptom of pain relieved in 3 cases (9%) in the first day after operation and disappeared in 3 days. Corneal epithelium of 9 cases ( 26%) were all healed within 1wk, 21 cases (60%) were all healed within 2wk, and 5 cases (14%) were all healed within 3wk. Following up for 6-18mo, there was no recurrence of symptoms or bulla. A small amount of tiny bubbles in the surrounding area appeared in 2 cases after 3 and 4wk. All patients had no neovascularization, and had smooth corneal surface. The amnions of 30 cases (86%) were thinning after 2mo, partially dissolved and absorped, not seen with the naked eye after 3mo. Thirty-four cases (97%) had no changes in vision, one case (3%) was from the light to front of the manual. After 2mo, corneal sensation decreased in 30 cases ( 86%) , corneal thickness increased from preoperative 788±35μm to 940±43μm. After 12mo, corneal thickness increased to 1060±27μm. Results of confocal microscopy: after 3mo, the number density of the trigeminal nerve fibers under corneal basement membrane reduced, shallow stromal cells became into fibrotic stroma, deep stroma was more loose, and cells swelled significantly. The number of endothelial cells reduced and form swelled more obviously compared with preoperatively. CONCLUSION: Corneal acupuncture combined with amniotic membrane transplantation can effectively control the symptoms of BK, prevent the recurrence of BK, and especially it is a simple, safe and practical way for patients with poor visual function.

11.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 666-670, 2014.
Article in Chinese | WPRIM | ID: wpr-294418

ABSTRACT

<p><b>OBJECTIVE</b>To study the efficacy of Xiaoyao Powder (XYP) combined with interferon alpha (IFN-alpha) in treating HBeAg positive chronic hepatitis B (CHB) patients and the effect on their quality of life (QOL).</p><p><b>METHODS</b>Totally 193 patients with HBeAg-positive CHB confirmed by liver biopsy were randomly assigned to 2 groups, Group A (94 cases) and Group B (99 cases). IFN-alpha1b was subcutaneously injected to patients in Group A at the dose of 50 microg, thrice per week. Those in Group B additionally took XYP. The therapeutic course for all was 24 weeks. Clinical efficacy was observed by assessing ALT restoration rate, HBeAg negative rate, HBeAg conversion rate, HBV DNA negative rate, complete response rate, partial response rate, and symptoms integral. The evaluation of QOL was performed by using chronic liver disease questionnaire (CLDQ) score. Adverse reaction occurrence rate was observed in the two groups.</p><p><b>RESULTS</b>Better effects were obtained in Group A on ALT restoration rate, HBeAg negative rate, HBV DNA negative rate, complete response rate, partial response rate, TCM symptoms integral, the total effective rate of TCM sysmptoms, CLDQ score, and adverse reaction rates, showing statistical difference when compared with Group B (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>XYP could elevate the efficacy of TCM symptoms of HBeAg-positive CHB patients and anti-viral effect, improve their QOL, and reduce adverse reaction of IFN-alpha.</p>


Subject(s)
Adult , Female , Humans , Male , Drugs, Chinese Herbal , Therapeutic Uses , Hepatitis B e Antigens , Hepatitis B, Chronic , Drug Therapy , Interferon-alpha , Therapeutic Uses , Quality of Life , Treatment Outcome
12.
Acta Pharmaceutica Sinica ; (12): 927-931, 2014.
Article in Chinese | WPRIM | ID: wpr-244993

ABSTRACT

This is to report the screening, extracting and validating antitumor components and compounds from Stellera chamaejasme L. under the case of discrete distribution of active data. In this work, different components from Stellera chamaejasme L. were collected by HPD macroporous resin and polyamide resin column, and their antitumor activity on A549 were tested by MTT assay. Activity results indicate that activity of components at 30-39 min is more potent than that of Stellera chamaejasme L. extract, and the activity of components at 33.97 min is equivalent to positive drug, cis-platinum at 100 microg x mL(-1), but with totally different mode of action. Under the case of discrete activity, the weight analysis is capable of screening active components and compounds from natural products.


Subject(s)
Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Cell Line, Tumor , Drug Screening Assays, Antitumor , Thymelaeaceae , Chemistry
13.
Acta Pharmaceutica Sinica ; (12): 911-916, 2013.
Article in English | WPRIM | ID: wpr-259530

ABSTRACT

Z-Ligustilide, a major phthalide isolated from a widely used traditional Chinese medicine Ligusticum chuanxiong, possesses various pharmacological activities including neuroprotective, anti-inflammatory, antiproliferative and vasorelaxing effects. However, it is unstable and inclined to degrade in natural conditions, which limits its study and application greatly. In this study, degradation behavior of Z-ligustilide and its degradation products stored at room temperature under direct sunlight were investigated and structure elucidated by HPLC-UV, UPLC-QTOF-MS and NMR. Z-ligustilide degradation and total five degradation products were generated and detected. Two degradation products were unequivocally identified as senkyunolide I and senkyunolide H by comparison with reference compounds. Another two degradation products were further isolated by semi-preparative HPLC and structure elucidated as (E)-6, 7-trans-dihydroxyligustilide and (Z)-6, 7-epoxyligustilide by 1H and 13C NMR, respectively. The degradation pathways of Z-ligustilide were finally proposed. Oxidation, hydrolysis and isomerization are the major degradation reactions.


Subject(s)
4-Butyrolactone , Metabolism , Benzofurans , Metabolism , Chromatography, High Pressure Liquid , Hydrolysis , Ligusticum , Chemistry , Magnetic Resonance Spectroscopy , Metabolic Networks and Pathways , Oxidation-Reduction , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
14.
China Journal of Chinese Materia Medica ; (24): 3647-3650, 2012.
Article in Chinese | WPRIM | ID: wpr-346890

ABSTRACT

<p><b>OBJECTIVE</b>To analyze and identify the chemical constituents in rat brain tissues after oral administration of Chuanxiong Rhizoma extracts.</p><p><b>METHOD</b>The dosed and blank rat brain tissues were analyzed by UPLC-Q-TOF-MS. Different peaks were observed in total ion chromatograms and then identified according to their retention time, accurate mass weight, MS and MS/MS data.</p><p><b>RESULT</b>After oral administration of Chuanxiong Rhizoma extracts, 3 compounds were absorbed into rat brain tissues through BBB. They were identified as senkyunolide I, senkyunolide A and ligustilide.</p><p><b>CONCLUSION</b>The study is helpful for interpreting effective substance of Ligusticum chuanxiong.</p>


Subject(s)
Animals , Female , Humans , Rats , Brain , Metabolism , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Pharmacokinetics , Ligusticum , Chemistry , Mass Spectrometry , Methods , Rats, Sprague-Dawley , Rhizome , Chemistry
15.
Chinese Journal of Virology ; (6): 143-149, 2010.
Article in Chinese | WPRIM | ID: wpr-297893

ABSTRACT

Based on the duck plague virus (DPV) UL35 gene sequence that our laboratory obtained (GenBank accession number EF643558), a pair of primers was designed using Oligo6.0 and primer5.0, then the UL35 gene was amplified from DPV CHv strain genomic DNA and cloned into the pMD18-T to construct a clone plasmid pMD18-T-UL35. After identification of the pMD18-T-UL35 by PCR amplification and restriction digestion, the fragment of the UL35 gene was subcloned into the prokaryotic expression vector pET-32a(+). The resultant recombinant plasmid pET-32a(+)-UL35 was then transformed into E. coli BL21 (DE3) strain and optimally-expressed under the induction of 1.0 mmol/L IPTG at 34 degrees C for 5 hours. SDS-PAGE analysis showed the recombinant protein (VP26) had a molecular weight of about 33KDa and accounted for 32.3% of total bacterial protein by gel scanning. The protein was then purified by Ni(2+)-affinity chromatography and used to immunize rabbit for producing the VP26 anti-serum and its antibody titer was up to 1:32 detected by agar diffusion reaction. After the IgG of the polyclonal antibodies was purified by High-Q anion-exchange chromatography, Western blot analysis indicated that the IgG had specific reaction with the VP26. Moreover, the subcellular localization detection was observed using immunofluorescence technique. The results showed that the specific fluorescences appeared relatively few in nucleus in 2 to 8 hours and increased gradually in 12 to 36 hours and eventually reached to the maximum, which aggregated in the spot region of the nucleus after the duck embryo fibroblast (DEF) were infected by DPV. However, there were only a small amount of specific fluorescences in the cytoplasm in 12 hours and increased with the extension of infection time in 24 to 48 hours. The specific fluorescences finally reached to the maximum in the cytoplasm in 72 hours. The results provided significant data for furthering the study on the function of DPV UL35 gene.


Subject(s)
Animals , Rabbits , Blotting, Western , Capsid Proteins , Chemistry , Genetics , Metabolism , Cell Nucleus , Metabolism , Cells, Cultured , Cloning, Molecular , Ducks , Virology , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Genetics , Fibroblasts , Cell Biology , Metabolism , Virology , Herpesviridae , Genetics , Metabolism , Microscopy, Fluorescence , Molecular Weight , Plasmids , Genetics , Polymerase Chain Reaction , Recombinant Proteins , Genetics , Allergy and Immunology , Metabolism
16.
National Journal of Andrology ; (12): 244-249, 2010.
Article in Chinese | WPRIM | ID: wpr-252822

ABSTRACT

<p><b>OBJECTIVE</b>The Ala499Val (C > T) and Lys939Gln (A > C) of the XPC gene are two potentially functional nonsynonymous polymorphisms, which affect the rate of DNA repair and might change XPC production and activity. This study aimed to explore the distribution of these two polymorphisms in the Chinese Han population and their relationship with male infertility.</p><p><b>METHODS</b>We genotyped the two polymorphisms of the XPC gene by the PCR-restriction fragment length polymorphism (PCR-RFLP) method in 318 infertile patients and 228 fertile male controls, detected the frequency of the alleles, and analyzed both the individual and the joint contribution of the two polymorphisms to male infertility.</p><p><b>RESULTS</b>For the Ala499Val (C > T) polymorphism, the frequencies of the CC, CT, and TT genotypes were significantly different in distribution between the patients and the controls (P = 0.020). Males with the TT genotype had a lower risk of male infertility than those with the CC genotype (adjusted OR = 0.49, 95% CI: 0.23-0.88), and even lower than those with both CC and CT genotypes (adjusted OR = 0.39, 95% CI: 0.22-0.71). The Lys939Gln (A > C) polymorphism was not related with male infertility. The combined genotype analysis showed that the individuals with 1-4 risk alleles had a significantly higher risk of male infertility (adjusted OR = 2.75, 95% CI = 1.50-5.04) than those with 0 risk allele.</p><p><b>CONCLUSION</b>The Ala499Val (C > T) polymorphism of the XPC gene is correlated with male infertility and may be a potential genetic risk factor for male infertility in the Chinese Han population.</p>


Subject(s)
Adult , Humans , Male , Alleles , Asian People , Genetics , Case-Control Studies , DNA Repair , DNA-Binding Proteins , Genetics , Gene Frequency , Genetic Predisposition to Disease , Genotype , Infertility, Male , Genetics , Polymorphism, Genetic , Risk Factors
17.
Chinese Journal of Microbiology and Immunology ; (12): 198-203, 2009.
Article in Chinese | WPRIM | ID: wpr-381039

ABSTRACT

Objective To investigate the regulation of protein kinase C(PKC) to the expression of β-1,4-galactusyhransferase- Ⅰ ( β-1,4-GalT- Ⅰ ) and the influence on cytoskeleton and adherence ability of human umbilical vein endothelial cells(HUVECs) when stimulated by lipopolysaccharide (LPS). Methods Cultured HUVECs were pretreated by various PKC inhibitors or phorbol 12-myristate 13-acctate( PMA), an excitomotor of PKC respectively for 30 min, then stimulated by LPS for 4 h. β-1,4-GalT-Ⅰ expression were detected by RT-PCR and Western blot, expression of β-1,4-galactosylated carbohydrate chains and cytoskeleton were assayed by immumofluorescence, and adherence ability of HUVECs was observed by endothelialmonocyte cell adherence test. Results Up-regulated expression of β-1,4-GalT- Ⅰ and β-1,4-galactosylated carbohydrate chains in HUVECs stimulated by LPS were suppressed by PKC inhibitors and increased by PMA. F-actin and β-1,4-GalT- Ⅰ were partly co-localized in HUVECs. PKC inhibitor inhibited the effect of LPS on the distribution of F-actin and β-1,4-GalT- Ⅰ. Adherence ability of HUVECs enhanced by LPS was significantly suppressed by PKC inhibitor. Conclusion PKC signal transduction pathway may participate in regulating β-1,4-GalT-Ⅰ expression in endothelial cells stimulated by LPS. Furthermore, polytypes of PKC may participate in this regulating process; PKC might regulate cytoskeleton reorganization and adherence ability of EC through β-1,4-GalT-Ⅰ during inflammation.

18.
Chinese Journal of Preventive Medicine ; (12): 299-304, 2009.
Article in Chinese | WPRIM | ID: wpr-242655

ABSTRACT

<p><b>OBJECTIVE</b>To examine the relationship between prepregnancy body mass index (BMI) and the risk of pregnancy-induced hypertension (PIH) in Chinese population.</p><p><b>METHODS</b>Data were collected in 6 counties/cities covered by Perinatal Health Care Surveillance System which was part of the Sino-American cooperative project on neural tube defects prevention established in 1992. The study population consisted of 83 159 women who attended premarital or preconception medical physical examination and delivered single live births with at least 20 gestational weeks from 1995 to 2000 in Jiaxing area. The Chi-square test was employed to test the difference in the rates of PIH among groups with different BMI. Multivariate logistic regression was conducted to examine the association between prepregnancy BMI and the risk of PIH.</p><p><b>RESULTS</b>The rate of PIH was 11.01% (9153/83 159; 95% CI: 10.79% - 11.22%). The rate of PIH among women with BMI < 18.5 kg/m(2), 18.5 - 22.9 kg/m(2), 23.0 - 24.9 kg/m(2), and > or = 25.0 kg/m(2) were 9.08% (1405/15 472; 95% CI: 8.63% - 9.54%), 10.82% (6389/59 054; 95% CI: 10.57% - 11.07%), 14.63% (943/6444; 95% CI: 13.78% - 15.52%), and 19.00% (416/2189; 95% CI: 17.38% - 20.71%), respectively, the difference was significant (chi(trend)(2) = 261.028, P = 0.000). Taking those with BMI 18.5 - 22.9 kg/m(2) as reference, the unadjusted RR for PIH was 0.82 (95% CI: 0.77 - 0.87) among women with BMI < 18.5 kg/m(2), 1.41 (95% CI: 1.31 - 1.52) among women with BMI 23.0 - 24.9 kg/m(2), and 1.93 (95% CI: 1.73 - 2.16) among women with BMI > or = 25.0 kg/m(2). After controlling for area, maternal age at delivery, educational level, occupation, parity, times of prenatal visit as well as the individual or family history of chronic hypertension, the estimated RR were 0.85 (95% CI: 0.80 - 0.90), 1.37 (95% CI: 1.27 - 1.47) and 1.88 (95% CI: 1.68 - 2.10), respectively.</p><p><b>CONCLUSION</b>High prepregnancy BMI could increase the risk of PIH.</p>


Subject(s)
Adult , Female , Humans , Pregnancy , Body Mass Index , Hypertension, Pregnancy-Induced , Epidemiology , Incidence , Logistic Models , Rural Population , Urban Population
19.
Chinese Journal of Oncology ; (12): 330-334, 2008.
Article in Chinese | WPRIM | ID: wpr-348100

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression variation and significance of Skp2 and p27(kip1) during the proliferation of lymphoma cell line Jurkat cells.</p><p><b>METHODS</b>The binding of p27(kip1) and Skp2 in Jurkat cells were detected by immunoprecipitation. Jurkat cells were treated with serum starvation and release synchronization. The expression variation and subcellular localization of p27(kip1) and Skp2 were detected by subcellular fractionation, Western blot and double immunofluorescence labelling.</p><p><b>RESULTS</b>The results of immunoprecipitation suggested that p27(kip1) and Skp2 could bind each other in Jurkat cells. During the proliferation of Jurkat cells, the protein expression of p27(kip1) decreased and intranuclear p27(kip1) decreased significantly, while the Skp2 protein increased and cytoplasmic Skp2 increased significantly.</p><p><b>CONCLUSION</b>During the proliferation of Jurkat cells, the increased cytoplasmic synthesis of Skp2 may speed up p27(kip1) degradation via the ubiquitin-proteasome pathway, then intranuclear p27(kip1) decreases significantly, leading to an increased cell cycling activity.</p>


Subject(s)
Humans , Cell Nucleus , Metabolism , Cell Proliferation , Cyclin-Dependent Kinase Inhibitor p27 , Metabolism , Cytoplasm , Metabolism , Jurkat Cells , Lymphoma, B-Cell , Metabolism , Pathology , Protein Binding , S-Phase Kinase-Associated Proteins , Metabolism
20.
Chinese Journal of Virology ; (6): 72-75, 2008.
Article in Chinese | WPRIM | ID: wpr-334845

ABSTRACT

Replication of duck plague virus(DPV) in artificially infected ducks were detected by in situ hybridization (ISH) which employed a 37bp oligonucleotide as probe designed according to DPV DNA sequence in GenBank. The results indicated that DPV DNA was detected in liver, intestine and bursa Fabricius at 4 h, in spleen and esophagus at 6h, in thymus at 12h post infection; DPV DNA in lung and kidney was detected only in dead ducks and no positive signal was detected in muscle, heart, cerebrum and pancreas. DPV DNA was distributed in cell nucleus and cytoplasm. Hepatocytes, sinus endodermal cells and Kuffer's cells were the mainly infected cell types in liver. DPV DNA was mainly detected in epithelium of villi, in lamina propria of intestinal villi of duodenum, in stratum spinosum of esophagus, and in epithelium, cortex, medulla of bursa Fabricius. The positive signals were mainly detected in medulla of thymus, lymphocytes and macrophages of spleen. The research suggests that ISH is a direct and specific method in detecting DPV DNA in paraffin sections and it's also a good method for virus diagnosis and DNA location of DPV.


Subject(s)
Animals , DNA, Viral , Ducks , Virology , In Situ Hybridization , Influenza A virus , Genetics , Physiology , Virus Replication
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