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1.
Article in Chinese | WPRIM | ID: wpr-1024117

ABSTRACT

Objective To analyze the cognition level of health care workers(HCWs)and the management status of various levels of medical institutions towards allergy reactions to commonly used antimicrobial agents.Methods HCWs and clinical pharmacists who were related to the diagnosis and treatment of antimicrobial agents in 14 medical institutions of city-level and autonomous prefectures in Gansu Province were randomly selected for a questionnaire survey.The survey contents included respondents'basic information,criteria for judging antimicrobial allergy,awareness on procedures related to antimicrobial allergy,and antimicrobial management level of different levels of medical institutions.Results A total of 8 670 valid questionnaires from HCWs were collected,including 3 300 phy-sicians,5 024 nurses and 328 pharmacists.160,775,2 123 and 5 612 HCWs were with senior,associate,interme-diate and junior professional titles,respectively.87.66%of the HCWs received relevant training on antimicrobial management in the past two years,the proportion of HCWs from different levels of medical institutions who have received training on antimicrobial management in the past two years was statistically significant different(x2=42.668,P<0.001).HCWs with senior professional titles had the highest proportion of receiving relevant training(93.75%),there was a statistically significant difference in the proportion of receiving antimicrobial management training among HCWs with different professional titles in the past two years(x2=69.782,P<0.001).50.98%of HCWs were not clear about penicillin allergy,and most of whom were with junior professional titles,accounting for 68.52%.25.19%of HCWs expressed uncertainty about whether patients with penicillin allergy could use cephalosporins,225 of whom were with associate professional titles,accounting for 29.03%of the total number of HCWs with associate profe-ssional titles.6.11%of HCWs had no experience in skin test procedure;46.94%of HCWs expressed that their medical institutions had no or unclear about whether their medical institutions had an antimicrobial allergy assess-ment team.Conclusion HCWs'judgment on allergy reactions to commonly used antimicrobial agents and aware-ness on antimicrobial application is not high enough,and the overall management level of antimicrobial allergy in all levels of medical institutions is poor.The popularity of antimicrobial allergy assessment teams is not high,and there is an urgent need to strengthen supervision,management,training,et al.

2.
Journal of Experimental Hematology ; (6): 1469-1474, 2023.
Article in Chinese | WPRIM | ID: wpr-1009998

ABSTRACT

OBJECTIVE@#To investigate a family with congenital dysfibrinogenemia, and analyze the risk of hemorrhage and thrombosis and blood transfusion strategies.@*METHODS@#Prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT) of the proband and her family members were detected by automatic coagulometer, fibrinogen (Fg) activity and antigen were detected by Clauss method and PT algorithm respectively. Meanwhile, thromboelastometry was analyzed for proband and her family members. Then, peripheral blood samples of the proband and her family members were collected, and all exons of FGA, FGB and FGG and their flanks were amplified by PCR and sequenced to search for gene mutations.@*RESULTS@#The proband had normal APTT and PT, slightly prolonged TT, reduced level of Fg activity (Clauss method). The Fg of the proband's aunt, son and daughter all decreased to varying degrees. The results of thromboelastogram indicated that Fg function of the proband and her family members (except her son) was basically normal. Gene analysis showed that there were 6233 G/A (p.AαArg35His) heterozygous mutations in exon 2 of FGA gene in the proband, her children and aunt. In addition, 2 polymorphic loci were found in the family, they were FGA gene g.9308A/G (p.AαThr331Ala) and FGB gene g.12628G/A (p.BβArg478Iys) polymorphism, respectively. The proband was injected with 10 units of cryoprecipitate 2 hours before delivery to prevent bleeding, and no obvious bleeding occurred during and after delivery.@*CONCLUSION@#Heterozygous mutation of 6233G/A (p.AαArg35His) of FGA gene is the biogenetic basis of the disease in this family with congenital dysfibrinogenemia.


Subject(s)
Humans , Child , Female , Fibrinogen/genetics , Pedigree , Afibrinogenemia/genetics , Mutation , Blood Transfusion
3.
Chinese Journal of Radiology ; (12): 364-369, 2023.
Article in Chinese | WPRIM | ID: wpr-992968

ABSTRACT

Objective:To explore the accuracy of artificial intelligence (AI) system based on deep learning in evaluating bone age of children with abnormal growth and development.Methods:The positive X-ray films of the left wrist of children with abnormal growth and development who were treated at the Affiliated Hospital of Guizhou Medical University from January 2020 to December 2021 were collected retrospectively. A total of 717 children were collected, including 266 males and 451 females, aged 2-18 (11±3) years. Based on Tanner Whitehouse 3 (TW 3)-RUS (radius, ulna, short bone) and TW3-Carpal (carpal bone) method, bone age was measured by 3 senior radiologists, and the mean value was taken as reference standard. The bone ages were independently evaluated by the AI system (Dr.Wise bone age prediction software) and two junior radiologists (physicians 1 and 2). The accuracy within 0.5 year, the accuracy within 1 year, the mean absolute error (MAE) and the root mean square error (RMSE) between the evaluation results and the reference standard were analyzed. Paired sample t-test was used to compare MAE between AI system and junior physicians. Intraclass correlation coefficient (ICC) was used to evaluate the consistency between AI system, junior physician and reference standard. The Bland-Altman diagram was drawn and the 95% consistency limit was calculated between AI system and reference standard. Results:For TW3-RUS bone age, compared with the reference standard, the accuracy within 0.5 year of AI system, physician 1 and physician 2 was 75.3% (540/717), 62.1% (445/717) and 66.2% (475/717), respectively. The accuracy within 1 year was 96.9% (695/717), 86.3% (619/717) and 89.1% (639/717), respectively. MAE was 0.360, 0.565 and 0.496 years, and RMSE was 0.469, 0.634 and 0.572 years, respectively. For TW3-Carpal bone age, compared with the reference standard, the accuracy within 0.5 year of AI system, physician 1 and physician 2 was 80.9% (580/717), 65.1% (467/717) and 71.7% (514/717), respectively. The accuracy within 1 year was 96.0% (688/717), 87.3% (626/717) and 90.4% (648/717), respectively. MAE was 0.330, 0.527 and 0.455 years, and RMSE was 0.458, 0.612, 0.538 years, respectively. Based on TW3-RUS and TW3-Carpal bone age, the MAE of AI system were lower than those of physician 1 and physician 2, and the differences were statistically significant ( P all<0.001). The evaluation results of AI, physician 1 and physician 2 were in good agreement with the reference standard (ICC all>0.950). The Bland-Altman analysis showed that the 95% agreement limits of AI system for assessing TW3-RUS and TW3-Carpal bone age were -0.75-1.02 years and-0.86-0.91 years, respectively. Conclusion:The accuracy of AI system in evaluating the bone age of children with abnormal growth and development is close to that of senior doctors, better than that of junior doctors, and in good agreement with senior doctors.

4.
Article in Chinese | WPRIM | ID: wpr-1015703

ABSTRACT

Metastasis and cell infiltration are the difficulties in the treatment of solid and lymphatic carcinoma and the main causes of disease recurrence and death. The migration of cancer cells is a prerequisite for tumor metastasis and invasion. The CXCL12-CXCR4 pathway plays an important role in the pathogenesis of solid tumors and leukemia. The interaction between CXCL12 and its receptor CXCR4 can activate multiple signaling pathways and regulate different physiological and pathophysiological processes. Thus, blocking CXCL12-CXCR4 binding and / or downstream pathways has clinical benefits in treating a variety of diseases and cancers. Although some CXCL12 and CXCR4 antagonists have been identified and have shown encouraging results in terms of antitumor activity, these drugs have not been widely used in clinical patients due to their serious toxic and side effects. There is an urgent need to develop novel CXCL12-CXCR4 axis antagonists for the treatment of tumors. Herein, we review the recent research progress of CXCR4 pathway in solid tumors and leukemia, and discuss the therapeutic value and future research direction of CXCR4 pathways in solid tumors and leukemia.

5.
Chinese Pharmaceutical Journal ; (24): 1169-1175, 2019.
Article in Chinese | WPRIM | ID: wpr-857963

ABSTRACT

OBJECTIVE: To establish a method for simultaneous determination of voriconazole and its main metabolite N-oxidized voriconazole in blood, and monitor the levels of voriconazole and N-oxidized voriconazole in leukemia children aged 2-12 years. METHODS: Agilent ZORBAX SB-C18 column (4.6 mm×250 mm,5 μm) was used, and the mobile phase was acetonitrile, methanol and ammonium acetate buffer (10 mmol•L-1, pH adjusted with acetic acid to 3.0)=27∶23∶50, the flow rate was 1 mL•min-1, and the wavelength was set at 262 nm. The sample was extracted with ethyl acetate.Carbamazepine was used as the internal standard. RESULTS: The calibration curves of voriconazole and N-oxidized voriconazole were linear in the range of 0.1-8.0 μg•mL-1, the lower limit of quantitation was 0.1 μg•mL-1, and the intra-assay and inter-assay precision RSDs were between 2% and 10%. The accuracy, method recovery and stability all met the requirements.Twenty-nine patients were monitored with a total of 100 blood samples.The medians (IQRs) of voriconazole concentration, N-oxidized voriconazole concentration, and ratio of N-oxidized voriconazole to voriconazole were 0.87 μg•mL-1 (0.32-1.65 μg•mL-1) and 1.15 μg•mL-1 (0.55-1.67 μg•mL-1) and 1.10(0.66-1.22), respectively. There was a correlation relationship between the concentrations of voriconazole and N-oxidized voriconazole (r=0.603 1, P<0.000 1). CONCLUSION: The method is simple, accurate, highly specific, and can simultaneously monitor the concentrations of voriconazole and N-oxidized voriconazole, thus can be used for the pharmacokinetic research of voriconazole and clinical drug monitoring.

6.
Article in Chinese | WPRIM | ID: wpr-816318

ABSTRACT

OBJECTIVE: To investigate the effect of HR-HPV positive,HR-HPV load and SCC-Ag positive on the recurrence of cervical cancer after radical resection.METHODS: The clinical data of cervical cancer patients who underwent radical resection of cervical cancer in People's Hospital of Zengcheng District from January 2010 to January2019 were retrospectively collected.The patients were followed up regularly,and the preoperative HR-HPV positive,loading,the amount of SCC-Ag expression were determined;the recurrence and metastasis of cervical cancer were analyzed,and the value of HR-HPV positive,HR-HPV loading and SCC-Ag in predicting the recurrence and metastasis of cervical cancer were analyzed.RESULTS: A total of 438 patients with cervical cancer were included.There was no significant difference in pathological type,or postoperative Meigs-Brunschwig pathological staging between the recurrence group(n=42)and the non-recurrence group(n=396)(P>0.05).The difference in the proportion of HRHPV positive(40/42 vs. 144/396),HR-HPV loading and SCC-Ag positive(34/42 vs. 64/396)was statistically significant between non-recurrence group and recurrence group(P0.05).Multivariate logistic regression analysis showed that distant metastasis,FIGO staging of cervical cancer,HR-HPV positive,and SCC-Ag were independent factors affecting cervical cancer recurrence(P<0.05).When predicting by individual indicator,the specificity and positive predictive value of HR-HPV positive for predicting cervical cancer recurrence were 99.18% and 95.23% at the highest,and the negative predictive value of HR-HPV was87.37% at the highest.When SCC-Ag was used to predict cervical cancer recurrence,the sensitivity was up to 33.33%.The sensitivity of combined prediction was 64.51%,the specificity was 99.46%,the positive predictive value was97.41%,and the negative predictive value was 94.44%.CONCLUSION: Distant metastasis,FIGO staging,HR-HPV positive,and SCC-Ag are independent factors affecting cervical cancer recurrence.The combination of HR-HPV positive,HR-HPV loading and SCC-Ag has certain value for predicting recurrence of cervical cancer,and the prediction value is the highest.

7.
International Eye Science ; (12): 414-417, 2019.
Article in Chinese | WPRIM | ID: wpr-719741

ABSTRACT

@#AIM: To analyze the early visual quality differences of femtosecond laser small incision matrix lens extraction(SMILE)versus femtosecond laser excimer laser <i>in situ</i> keratomileusis(LASIK).<p>METHODS: From February 2017 to February 2018, 78 patients(including 156 eyes)with SMILE myopia were enrolled in our hospital. Eighty patients(160 eyes)who underwent LASIK myopia in the same period were selected. All patients were followed up for three times. Months were observed for surgical outcomes, high-order aberrations, and contrast sensitivity(CS)status.<p>RESULTS: At 1mo and 3mo postoperatively, the central corneal thickness change rate in the SMILE group was(-14.48±2.67)%,(-13.54±2.90)% lower than that of LASIK(-17.92±2.85)%,(-15.63±2.71)%, and intraocular pressure changes. The rates were(-27.08±3.64)% and(-24.41±3.28)% were lower than those in the LASIK group(-29.26±3.81)% and(-27.01±3.62)%(<i>P</i><0.05). The total high-order aberrations of the SMILE group were the sputum difference was lower than that of LASIK group(<i>P</i><0.05). The SMILE group values were(1.86±0.21),(1.52±0.23)and(0.91±0.14)when the spatial frequency of glare CS was 1.5, 12.0 and 18.0 at 1mo after operation was higher than the LASIK group(1.71±0.20),(1.41±0.25),(0.81±0.12)(<i>P</i><0.05).<p>CONCLUSION: SMILE has a lower effect on patients with higher-order aberrations than LASIK, and the early visual quality is better.

8.
Article in Chinese | WPRIM | ID: wpr-231625

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of Erigeron Breviscapus (EB) at different concentrations and different intervention time points on the mRNA and protein expression of OPG/RANKL/RANK in MG63 osteoblast-like cells and RAW264. 7 pre-osteoclast cells cultured in vitro, thus exploring roles EB played in bone rebuilding and its mechanisms.</p><p><b>METHODS</b>MG63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro. The 3rd passage cells were divided into the control group and different experimental groups. Total RNA and protein were respectively isolated from cells treated with different concentrations of EB (0, 0.001, 0.01, 0.1, and 1.0 mg/mL) for 48 h. Meanwhile, the protein was extracted from 0 and 1 mg/mL EB groups at 12, 24, and 48 h respectively. Expression of OPG mRNA and RANKL mRNA in MG63 osteoblast-like cells, and expression of RANK mRNA in RAW264.7 pre-osteoclast cells were detected by semi-quantitative RT-PCR. Expression of OPG protein and RANKL protein in MG63 osteoblast-like cells, and expression of RANK protein in RAW264. 7 pre-osteoclast cells were detected by Western blot.</p><p><b>RESULTS</b>Along with increased EB concentration, expression of OPG mRNA and protein in MG63 osteoblast-like cells was gradually lowered (P < 0.05) after 48-h intervention of EB, the expression of RANKL mRNA and protein in MG63 osteoblast-like gradually increased (P < 0.05); the expression of RANK mRNA in RAW264.7 pre-osteoclast cells increased (P < 0.05). But the expression of RANK mRNA was slightly lower in the 0.1 mg/mL EB group than in the 0.01 mg/mL EB group, and the expression of RANK protein in RAW264.7 pre-osteoclast cells gradually increased (P < 0.05). After treatment with 1 mg/mL EB for 12, 24, 48 h, the expression of OPG protein in MG63 osteoblast-like cells gradually decreased as time went by (P < 0.05), and the expression of RANKL protein in MG63 osteoblast-like and RANK protein in RAW264.7 pre-osteoclast cells gradually increased (P < 0.05). The expression of RANKL protein in RAW264.7 pre-osteoclast cells increased as time went by (P < 0.05).</p><p><b>CONCLUSION</b>EB could inhibit the expression of OPG in osteoblasts in a dose- and time-dependent manner, promote the expression of RANKL in osteoblasts and the secretion of RANK in pre-osteoclast, indicating EB might play roles in promoting bone resorption.</p>


Subject(s)
Animals , Humans , Mice , Cell Differentiation , Cell Line , Drugs, Chinese Herbal , Pharmacology , Erigeron , Osteoblasts , Metabolism , Osteoclasts , Metabolism , Osteoprotegerin , Metabolism , RANK Ligand , Metabolism , RNA, Messenger , Genetics , Receptor Activator of Nuclear Factor-kappa B , Metabolism
9.
Article in Chinese | WPRIM | ID: wpr-233695

ABSTRACT

<p><b>OBJECTIVE</b>To study hepatitis B virus (HBV) expression in 3 hepatocytes infected with recombinant adenovirus containing 1.2-copy HBV DNA.a</p><p><b>METHODS</b>A chicken hepatoma cell line and two human hepatocytes were infected by the recombinant adenovirus containing 1.2-copy HBV DNA at 25 pfu/cell. HBV-specific mRNA was detected by RT-PCR 3 days after the infection, and HBsAg and HBeAg were detected by ELISA and HBV DNA by real-time PCR daily after the infection.</p><p><b>RESULTS</b>HBV mRNA expression was detected in all the 3 cells after recombinant adenovirus infection, and the quantities of HBV DNA and HBV antigens in the culture supernatant increased with the passage of time.a</p><p><b>CONCLUSION</b>Infection with the recombinant adenovirus containing 1.2-copy HBV DNA can mediate HBV infection in the 3 cells in vitro.</p>


Subject(s)
Animals , Humans , Adenoviridae , Genetics , Cell Line, Tumor , Culture Media, Conditioned , Metabolism , DNA, Recombinant , Genetics , DNA, Viral , Genetics , Metabolism , Gene Expression , Hepatitis B Antigens , Metabolism , Hepatitis B virus , Hepatocytes , Metabolism , Virology , Reverse Transcriptase Polymerase Chain Reaction
10.
Chinese Journal of Hepatology ; (12): 422-424, 2007.
Article in Chinese | WPRIM | ID: wpr-230577

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the relationship of virological breakthrough and production of neutralizing anti-interferon antibody (NAb) in chronic hepatitis B patients treated with recombinant interferon-alpha (rIFN-alpha).</p><p><b>METHOD</b>Four hundred eighty-five patients with histological proven chronic hepatitis B were treated with 5 MU recombinant interferon-alpha 1b (rIFN-alpha1b) thrice weekly for 6-37 months (median 10). Serum HBV DNA, HBeAg and NAb levels of the patients were detected by fluorescent-quantitative PCR, enzymoimmunoassay and antiviral neutralizing biological assay respectively during the therapy.</p><p><b>RESULTS</b>Virological breakthrough occurred in 66 patients (13.6%), and NAb was found in 98 patients (20.2%) of the total 485 patients. The rate of NAb positivity was higher in patients with viral breakthrough than those without it (68.2%, 45/66, vs 12.6%, 53/419, chi(2)=109.06, P < 0.01), and viral breakthrough occurred more in patients with positive NAb than with negative NAb (45.9%, 45/98, vs 5.4%, 21/387, chi(2)=109.06, P < 0.01). The time of the viral breakthrough occurrence and the time of NAb production had a significant correlation (P < 0.01). The occurrence of viral breakthrough was also influenced by the age of patients (P < 0.05) and HBeAg status (P < 0.01) before they were treated.</p><p><b>CONCLUSION</b>Viral breakthrough occurred in 13.6% of our 485 chronic hepatitis B patients treated with recombinant interferon-alpha. Their viral breakthrough and production of NAb production had a significant correlation.</p>


Subject(s)
Adult , Female , Humans , Male , Young Adult , Antibodies, Neutralizing , Hepatitis B Antibodies , Hepatitis B virus , Allergy and Immunology , Hepatitis B, Chronic , Drug Therapy , Virology , Interferon Type I , Therapeutic Uses , Recombinant Proteins
11.
Chinese Journal of Biotechnology ; (12): 902-906, 2007.
Article in Chinese | WPRIM | ID: wpr-276189

ABSTRACT

Carbon source plays an important role in the constitutive expression of foreign proteins in Pichia pastoris. In present study, glucose , glycerol , methanol and oil acid, was used respectively as the only carbon source to constitutively express hAS in Pichia pastoris GS115 (pGAP9K-AS)in shaking flask. The result shows that oleic acid is the best (163 mg/L) compared with glycerol (83mg/L), glucose (76 mg/L)and methanol (57 mg/L). Since oleic acid is insoluble in water, glycerol was used as the carbon source in the high-density cell culture of GS115 (pGAP9K-AS) in a 30 liter bioreactor and 169 mg/L of angiostatin was obtained after 48h of culture. The expressed angiostatin is immunologically active as shown by Western blotting. The recombinant hAS inhibits bFGF induced CAM angiogenesis and suppresses the growth of B16 melanoma in C57BL/6J mice. The tumor inhibition rate is 90% after 12 days of treatment. Statistics analysis revealed that the tumor volume difference of mice between the hAS group and PBS group is prominent (P < 0.01).


Subject(s)
Animals , Humans , Mice , Angiogenesis Inhibitors , Genetics , Therapeutic Uses , Angiostatins , Genetics , Therapeutic Uses , Bioreactors , Microbiology , Culture Media , Pharmacology , Fermentation , Glycerol , Pharmacology , Melanoma, Experimental , Drug Therapy , Mice, Inbred C57BL , Pichia , Genetics , Metabolism , Recombinant Proteins , Genetics , Therapeutic Uses
12.
Article in Chinese | WPRIM | ID: wpr-283126

ABSTRACT

<p><b>OBJECTIVE</b>o study the replication of hepatitis B virus (HBV) in HepG2 cells infected with Ad-1.2 HBV.</p><p><b>METHODS</b>HepG2 cells were transfected with adenovirus containing 1.2 copies of HBV DNA. The expression of HBV antigens were detected in the culture medium by means of enzyme-linked immunosorbent assay (ELISA), and the covalently closed circular DNA (cccDNA) in the cells was extracted with plasmid extraction kit and detected by real-time PCR with selective primer after treatment with mung bean nuclease.</p><p><b>RESULTS</b>HBsAg, HBeAg and HBV cccDNA were all detected in HepG2 cells after tranfection with Ad-1.2 HBV. HBV cccDNA was detected 1 day after the infection, reaching the peak level 4 days after infection.</p><p><b>CONCLUSION</b>Ad-1.2 HBV-infected cells can serve as the model for screening and evaluation of antiviral agents.</p>


Subject(s)
Humans , Adenoviridae , Genetics , Calibration , Cell Line, Tumor , DNA, Complementary , Genetics , Metabolism , DNA, Viral , Genetics , Metabolism , Hepatitis B Surface Antigens , Metabolism , Hepatitis B e Antigens , Metabolism , Hepatitis B virus , Genetics , Allergy and Immunology , Metabolism , Physiology , Polymerase Chain Reaction , Time Factors , Transfection , Virus Replication
13.
Article in Chinese | WPRIM | ID: wpr-283154

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the related to relapse of chronic hepatitis B (CHB) after recombinant interferon-alpha (rIFN-alpha) treatment.</p><p><b>METHODS</b>This investigation involved 523 pathologically confirmed CHB patients including 403 HBeAg-positive and 120 HBeAg-negative patients, who were treated with 5 MU rIFN-alpha subcutaneously thrice a week for 6-25 months. For each patient, serum alanine aminotransferase (ALT) was measured biochemically, serum HBV DNA level detected with quantitative fluorescent PCR, and HBeAg level with enzyme immuoassay every 1-3 months during therapy and every 3-6 months during the follow-up period.</p><p><b>RESULTS</b>Early response to rIFN-alpha treatment was observed in 302 (57.7%) patients at the end of treatment, among whom 39.4% (119/302) suffered relapse during the follow-up for 39.2-/+21.5 months. Age, HBeAg status before treatment, and follow-up duration were the predictive factors for post-treatment relapse. The mean age of patients with CHB relapse was significantly higher than that of the sustained responders (P<0.001), and the relapse rates in HBeAg-negative group (55.8%, 43/77) were significantly higher than that in HBeAg-positive group (33.8%, 76/225) at the end of follow up (P<0.001). The relapse rate and accumulative relapse rates at each year during the follow-up (for 5 years as the longest) differed significantly (P<0.001, P=0.000), but the accumulative relapse rates differed little between the years after the initial 2 of the follow-up (P=0.670). The relapse was not related to the patient's gender, pretreatment serum ALT, HBV DNA, grade of liver inflammation, stage of liver fibrosis, or duration of treatment. In HBeAg-positive patients, however, the mean HBV DNA was significantly higher in relapse group than in sustained response group (P=0.017).</p><p><b>CONCLUSION</b>Age, pretreatment HBeAg status, and follow-up duration are independent predictive factors for post-treatment CHB relapse. In HBeAg positive patients, pretreatment serum HBV DNA is also one of the risk factors for relapse.</p>


Subject(s)
Adult , Female , Humans , Male , Age Factors , Alanine Transaminase , Blood , DNA, Viral , Blood , Follow-Up Studies , Hepatitis B e Antigens , Blood , Hepatitis B, Chronic , Blood , Drug Therapy , Therapeutics , Interferon-alpha , Therapeutic Uses , Logistic Models , Recurrence , Treatment Outcome
14.
Article in Chinese | WPRIM | ID: wpr-245767

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the efficacy and investigate the influencing factors of the interferon (IFN) retreatment for patients with chronic hepatitis C relapsed after a previous IFN treatment.</p><p><b>METHODS</b>A retrospective study was designed to analyze the retreatment with IFN of 60 relapsed chronic hepatitis C patients. All patients were from a randomized, opened and multi-center clinical trial about the efficacy and security of PEG-IFNalpha-2a compared to CIFNalpha-2a in the treatment of chronic hepatitis C in China. There were 35 patients treated with PEG-IFNalpha-2a and 25 with CIFNalpha-2a. The main parameter to evaluate the efficacy was sustained viral response (SVR) rate. The influence of viral concentration in serum, genotype and drug categories on the responses to IFN were analyzed.</p><p><b>RESULTS</b>For all the patients, the end of treatment virus response (ETVR) and SVR rates were 55.00% and 35.00% respectively. ETVR rate of PEG-IFNalpha-2a was significantly higher than that of CIFNalpha-2a (74.29% and 28.00% respectively, P < 0.01). SVR rate of PEG-IFNalpha-2a was also markedly higher than that of CIFNalpha-2a (45.71% and 20.00% respectively, P < 0.05). However, there was no significant difference between the high and low viral load groups. Among the patients with genotype 1, ETVR and SVR rates of PEG-IFNalpha-2a (75.00%, 45.83%) were significantly higher than those of CIFNalpha-2a (22.22%, 11.11%), (P < 0.01, P < 0.05 respectively), but in patients with genotype non-1, there were no such differences between the two groups.</p><p><b>CONCLUSION</b>Some relapsed patients were not responsive to the IFN retreatment. The efficacy of PEG-IFNalpha-2a was superior to CIFNalpha-2a. The conventional IFN was not suggested to be used in the relapsed cases with genotype 1. The viral load was not associated with the efficacy of IFN retreatment.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Antiviral Agents , Therapeutic Uses , Hepatitis C, Chronic , Therapeutics , Interferon-alpha , Therapeutic Uses , Interferon-beta , Interferons , Therapeutic Uses , Polyethylene Glycols , Therapeutic Uses , Recombinant Proteins , Recurrence , Retrospective Studies
15.
Chinese Journal of Biotechnology ; (12): 451-456, 2006.
Article in Chinese | WPRIM | ID: wpr-286268

ABSTRACT

Human tumstatin(hTumstatin)cDNA was amplified from recombinant plasmid pET-3c-tum, cloned in frame with the signal sequence in yeast vector pPICZalphaA and transformed into Pichia pastoris GS115 by electroporation. The expression of hTumstatin in GS115(pPICZalpha-tum)was then induced by methanol and secreted into the culture medium, with a yield of 25mg/L as shown by SDS-PAGE and Western blotting. The expressed hTumstatin was purified to more than 85% purity using a simple one-step SP-Sepharose cation exchange chromatography. The MTT and chick chorioallantoic membrane assay showed that the yeast produced hTumstatin could inhibit the proliferation of human umbilical vein endothelial cells and the neovascularization induced by bFGF. Hoechst 33258 fluorescent staining also demonstrated the apoptotic change in endothelial cellular nuclear morphology.


Subject(s)
Humans , Angiogenesis Inhibitors , Metabolism , Autoantigens , Genetics , Metabolism , Cell Proliferation , Cells, Cultured , Collagen Type IV , Genetics , Metabolism , DNA, Complementary , Genetics , Electroporation , Endothelial Cells , Cell Biology , Pichia , Genetics , Metabolism , Recombinant Proteins , Genetics , Metabolism , Umbilical Cord , Cell Biology
16.
Article in Chinese | WPRIM | ID: wpr-282932

ABSTRACT

<p><b>OBJECTIVE</b>To provide an cell model of immortalized lymphoblstoid B-cell lines for studying the biological characteristics of full-length hepatitis B virus (HBV) genome carrying the hot-spot mutations V60, G87, and L97.</p><p><b>METHODS</b>V60, G87, and L97 mutation points were introduced into HBV p3.8 II plasmid containing 1.2 copy of HBV genome by means of site-directed mutagenesis. The HBV genome was amplified by PCR from p3.8 II and p3.8 II-V60, G87, L97 plasmid, and the PCR product was inserted into EBO-plpp eukaryotic expression vector. The recombinant vectors and the EBO-plpp vector were transfected into immortalized human lymphoblasts with lipofectamine 2000 and selected with hygromycin. Steady expression of the target genes was determined by RT-PCR, Western blotting and microparticle enzyme immunoassay.</p><p><b>RESULTS</b>DNA sequence analysis indicated that the desired mutation was introduced into wild-type HBV DNA. HBsAg, HBeAg and HBcAg could be detected in EBO-HBV-transfected cell lysate or culture supernatant.</p><p><b>CONCLUSION</b>Transfectants that stably express HBV mutant antigen may provide a cell model to study the biological characteristics of HBV carrying hot-spot mutation in vitro.</p>


Subject(s)
Humans , B-Lymphocytes , Cell Biology , Virology , Base Sequence , Blotting, Western , Cell Line, Transformed , Cell Transformation, Viral , DNA, Viral , Genetics , Eukaryotic Cells , Metabolism , Gene Expression Regulation, Viral , Genetic Vectors , Genome, Viral , Genetics , Hepatitis B Core Antigens , Genetics , Metabolism , Hepatitis B virus , Genetics , Metabolism , Molecular Sequence Data , Point Mutation , Reverse Transcriptase Polymerase Chain Reaction , Transfection
17.
Chinese Journal of Hepatology ; (12): 565-568, 2006.
Article in Chinese | WPRIM | ID: wpr-341300

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the relationship between hepatitis C virus (HCV) genotype, serum viral load and ALT levels, and the factors associated with the viral relapse after IFN treatment in patients with chronic hepatitis C.</p><p><b>METHODS</b>The HCV RNA levels were determined with Cobas Amplicor Monitor Test, version 2.0, and HCV genotypes were examined by means of PCR products of 5' NTR digested with restriction endonucleases. The patients with chronic hepatitis C were treated with PEG-IFN alpha -2a and Roferon-A for 24 weeks. Those with a viral response after 24 week treatment were followed for an additional 24 weeks. The association of clinical characteristics, such as sex, age, the way of the HCV infection, IFN treatment history and platelet counts, and the HCV genotype, virus load and medicine used for the viral relapse after IFN treatment were analyzed.</p><p><b>RESULTS</b>Of the 208 chronic hepatitis C patients, the ALT levels were not related to HCV RNA levels (r = 0.093, P > 0.05). No difference of ALT levels between HCV genotypes was found, and the HCV RNA load was also of no difference between HCV genotype 1 patients and non 1 patients. Of the 119 patients with viral response after 24 week treatment, 58 cases (48.7%) relapsed after another 24 week's follow-up. Relapse was not significantly related to the clinical characteristics, such as sex, age, mode of the infection, treatment history of IFN, AST/ALT ratio, platelet counts and the baseline viral load. Among patients with genotype 1 virus, the relapse rate was significantly higher than those patients with non-genotype 1 virus (54.5% vs 32.1%, P=0.039). The relapse rate after PEG-IFN alpha -2a treatment was lower than that of Roferon-A treatment (47.0% vs. 52.8%), but not significantly.</p><p><b>CONCLUSION</b>The viral relapse of chronic hepatitis C patients after IFN treatment was significantly associated with the genotypes of the HCV.</p>


Subject(s)
Female , Humans , Male , Middle Aged , Antiviral Agents , Therapeutic Uses , Genotype , Hepacivirus , Genetics , Hepatitis C, Chronic , Drug Therapy , Virology , Interferon-alpha , Therapeutic Uses , RNA, Viral , Blood , Recombinant Proteins , Recurrence , Treatment Outcome , Viral Load
18.
Chinese Medical Journal ; (24): 56-61, 2005.
Article in English | WPRIM | ID: wpr-257324

ABSTRACT

<p><b>BACKGROUND</b>It is still unclear whether viral genetic variability influences response to interferon (IFN)-alpha treatment. Recent reports suggest that IFN-alpha effects may be associated with hepatitis B virus (HBV) post-transcriptional regulation. This study was designed to explore the heterogeneity of HBV post-transcriptional regulatory elements (HPRE) and the relationship between the diversity of HPRE and the response to IFN-alpha treatment.</p><p><b>METHODS</b>The HPRE sequences from 31 Chinese patients infected with HBV were determined by directly sequencing of polymerase chain reaction (PCR) product, and comparing them to those from Caucasian patients. Subsequently, eukaryotic expression vectors containing HPRE at various points were constructed and transfected into HepG2 cells, which were then exposed to recombinant human cytokines.</p><p><b>RESULTS</b>The T to C point mutation at nt 1504 and the C to T (G) at nt 1508 in HPRE were found in 21 and 19 patients with chronic hepatitis B, respectively; the C to T point mutation at nt 1509 was found in 17 patients. These point mutations did not exist in the HPRE of the Caucasian patients. The activity of the CAT gene obviously increased in the case of T to C point mutation at nt 1504, but did not change in the case of the C to T (G) mutations at nt 1508 and 1509. The activity of the CAT gene at these point mutations of HPRE could be inhibited by IFN-alpha/gamma and tumor necrosis factor (TNF)-alpha except for the point mutations at nt 1508 of HPRE which may escape the suppression role of IFN-alpha on HPRE.</p><p><b>CONCLUSIONS</b>There are point mutations between the HPRE of Chinese and Caucasian HBV patients, which might be correlated with response to IFN-alpha. The variation of HPRE might affect the function of HPRE and influence the regulative function of IFN-alpha other than that of IFN-gamma or TNF-alpha on HPRE.</p>


Subject(s)
Humans , Chloramphenicol O-Acetyltransferase , Metabolism , Genes, Regulator , Hepatitis B virus , Genetics , Hepatitis B, Chronic , Drug Therapy , Virology , Interferon-alpha , Pharmacology , Interferon-gamma , Pharmacology , Plasmids , Point Mutation , Tumor Necrosis Factor-alpha , Pharmacology
19.
Chinese Journal of Hepatology ; (12): 488-490, 2005.
Article in Chinese | WPRIM | ID: wpr-348758

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of genotypes of HBV and HBeAg on the response to PEG-interferon alpha (PEG-IFN) in chronic hepatitis B (CHB) patients.</p><p><b>METHODS</b>PCR-RFLP and S gene sequencing were conducted in 42 CHB patients.</p><p><b>RESULTS</b>The sustained response (SR) rates were 66.7% in genotype B and 27.3% in genotype C group. The P value was 0.039 by the Pearson Chi-square test, while it was 0.06 by the Fisher's exact test. The results suggested a trend that patients with genotype B HBV compared to genotype C had better SR to PEG-IFN therapy, although the difference was not significant. Results also showed that SR rate in patients with HBeAg-negative CHB (7/8 87.5%) was significantly higher than that in HBe+ CHB patients (8/21 38.1%, P < 0.05).</p><p><b>CONCLUSION</b>Our results indicate that HBV genotype and HBeAg, especially the later, are main factors for predicting PEG-IFN therapy response in CHB patients.</p>


Subject(s)
Adult , Female , Humans , Male , Antiviral Agents , Therapeutic Uses , Genotype , Hepatitis B e Antigens , Blood , Hepatitis B virus , Genetics , Allergy and Immunology , Hepatitis B, Chronic , Drug Therapy , Virology , Interferon-alpha , Therapeutic Uses , Polyethylene Glycols , Therapeutic Uses , Recombinant Proteins , Treatment Outcome
20.
Article in Chinese | WPRIM | ID: wpr-281809

ABSTRACT

<p><b>OBJECTIVE</b>To study the mechanism of hepatitis B virus infected patients who is negative for HbsAg.</p><p><b>METHODS</b>DNA sequences of 46 patients were analyzed. In these patients, HBsAg was negative but HBV DNA was positive and six new HBsAg variants were identified. Four of the six variants were combined point mutants and two were insertion variants. These S genes were subcloned into eukaryotic expression vector EBO-plpp, and the recombinant eukaryotic expression plasmids were transfected into COS7 cells. Cell lines expressing mutant type HBsAg were obtained. The supernatants were detected by ELISA and RIA.</p><p><b>RESULTS</b>Only the two-amino acid-insertion variants could be detected and the others failed to react with polyclonal and monoclonal antibodies against HbsAg.</p><p><b>CONCLUSION</b>The results indicated that the point mutations and insertions may result in a conformational change of the S gene, which affect HBsAg antigenicity, suggesting a possible relationship between the variants and the negative conversion of HBsAg of the patients.</p>


Subject(s)
Animals , Humans , Antigenic Variation , COS Cells , Chlorocebus aethiops , Hepatitis B Surface Antigens , Genetics , Allergy and Immunology , Hepatitis B virus , Genetics , Allergy and Immunology , Hepatitis B, Chronic , Allergy and Immunology , Virology , Plasmids , Genetics , Point Mutation , Transfection
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