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Disorders of gastrointestinal motility (DGIM) are very common diseases in the department of gastroenterology and the prevalence is increasing gradually. The pathogenesis of DGIM is complex and closely related to Cajal interstitial cells of gastrointestinal tract, brain-intestinal axis and intestinal microecology. So far, a variety of diagnostic techniques have emerged, including esophageal 24 h pH monitoring and impedance analysis, electrogastrography, radionuclide scanning, ultrasound, 13C gastric emptying breath test, hydrogen breath test, defecation contrast, X-ray marker method, high resolution manometry and wireless motility capsules. According to the different pathogenesis of DGIM, treatment methods emerge in an endless stream, such as adjustment of lifestyle, drugs and surgical treatment and so on. Individual treatment should be provided clinically for different patients.
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【Objective】 To study the current situation of apheresis platelets collection in various regions of Gansu province by comparing and analyzing relevant data from blood stations in 14 prefecture-level cities of Gansu province. 【Methods】 The units of collected platelets and rate of double-dose collection in 13 regional blood stations and 1 provincial blood center from 2016 to 2020, as well as the clinical supply and demand was statistically analyzed. 【Results】 From 2016 to 2020, the total units of platelets collected by 13 blood stations and 1 blood center in Gansu increased from 11 255 U to 15 270 U, with the increase rate at 35.7% in 5 years, and mainly were collected by the provincial blood center (74.57%, 50 253/ 67 392). Although the rate of double-dose collection in the province showed a steady upward trend, only 3 blood stations realized annual double-dose collection more than 20%. There was still a gap of about 10% between supply and clinical needs. 【Conclusion】 Although the number of platelet collections and units in each blood station in Gansu is on the rise in general, the units collected varies in each blood station. Therefore, further measures need to be taken from the aspects of publicity, recruitment, optimizing the collection process, improving the rate of double-dose collection, retention of regular blood donors and regional coordination to increase the collection units, narrow down the regional gap and ensure the balance between supply and demand.
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【Objective】 To analyze the comprehensive factors causing adverse reactions to apheresis platelet donation(ARAPD), so as to provide references for effective prevention of ARAPD. 【Methods】 The 272 cases of ARAPD from 2012 to 2019 in Lanzhou were statistically analyzed, and factors that induced ARAPD were studied. Statistical analysis were performed according to the gender, nationality, occupation, age, weight, donation units, and number of donations. 【Results】 As to the factors inducing ARAPD, anticoagulant reactions accounted for the first(32.4%, 88/272). Women and students were prone to develop ARAPD. Among all age groups, 18~25 years old were most likely to develop ARAPD(53.68%, 10 572/35 265). The incidence of ARAPD were significantly different by ages and weights(P<0.05), and donors with lighter weight were more prone to develop ARAPD(P<0.05). The incidence of ARAPD were also significantly different between first-time and repeated donors(P<0.05), but not among the donation units. 【Conclusion】 The anticoagulant reactions are the leading reason for ARAPD. For female, student, young, light-weight, and first-time blood donors, special attention should be paid and corresponding interventions taken to them.
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Objective@#To evaluate the disinfection efficacy of peracetic acid disinfectant (type Ⅲ ) on gastrointestinal endoscopy.@*Methods@#Endoscopes were disinfected respectively by 2% glutaraldehyde (GA group) and peracetic acid disinfectant (type Ⅲ ) (PAA group) according to the process by the 2016 version of "Regulation for cleaning and disinfection technique of flexible endoscope" , and then samples were collected through biopsy channel at the specified steps. The bacterial count and pathogenic bacteria of these samples were detected. Hepatitis B virus surface antigen (HBsAg), hepatitis C virus (HCV) antibody and Treponemia pallidum antibody (TP-Ab) were detected by chemiluminesent microparticle immunoassay (CMIA) in the PAA group. The PAA group were continuously sampled for 5 days.@*Results@#A total of 56 gastroscopes and 16 colonoscopes were disinfected in the GA group, and 46 gastroscopes and 15 colonoscopes were disinfected in the PAA group. Compared with pre-disinfection, the bacterial count was both significantly reduced in the two groups after disinfection (P<0.05). The qualified rate of gastroscopes disinfection and total qualified rate of the PAA group were higher than those of the GA group [the qualified rate of gastroscopes: 97.83% (45/46) VS 92.86% (52/56), P>0.05; total qualified rate: 98.36% (60/61) VS 94.44% (68/72), P>0.05]. The qualified rate of colonoscopes in the two groups were both 100.00% (15/15, 16/16). After disinfecting by peracetic acid disinfectant (Type Ⅲ), HBsAg, anti-HCV and TP-Ab were negative. There were no significant differences on colonies number at different steps in a 5-day continuous sampling (P>0.05).@*Conclusion@#Peracetic acid disinfectant (type Ⅲ) can provide a satisfied disinfectant effect, and be applied in clinic to meet the requests of high-level disinfection for gastrointestinal endoscopy.
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Objective To evaluate the disinfection efficacy of peracetic acid disinfectant (type Ⅲ ) on gastrointestinal endoscopy. Methods Endoscopes were disinfected respectively by 2％ glutaraldehyde (GA group)and peracetic acid disinfectant (type Ⅲ )(PAA group)according to the process by the 2016 version of "Regulation for cleaning and disinfection technique of flexible endoscope",and then samples were collected through biopsy channel at the specified steps. The bacterial count and pathogenic bacteria of these samples were detected. Hepatitis B virus surface antigen (HBsAg),hepatitis C virus (HCV)antibody and Treponemia pallidum antibody (TP-Ab) were detected by chemiluminesent microparticle immunoassay (CMIA)in the PAA group. The PAA group were continuously sampled for 5 days. Results A total of 56 gastroscopes and 16 colonoscopes were disinfected in the GA group,and 46 gastroscopes and 15 colonoscopes were disinfected in the PAA group. Compared with pre-disinfection,the bacterial count was both significantly reduced in the two groups after disinfection (P<0. 05). The qualified rate of gastroscopes disinfection and total qualified rate of the PAA group were higher than those of the GA group [the qualified rate of gastroscopes:97. 83％ (45/ 46)VS 92. 86％ (52/ 56),P>0. 05;total qualified rate:98. 36％ (60/ 61)VS 94. 44％ (68/ 72),P> 0. 05]. The qualified rate of colonoscopes in the two groups were both 100. 00％(15/ 15,16/ 16). After disinfecting by peracetic acid disinfectant (Type Ⅲ),HBsAg,anti-HCV and TP-Ab were negative. There were no significant differences on colonies number at different steps in a 5-day continuous sampling (P>0. 05). Conclusion Peracetic acid disinfectant (type Ⅲ)can provide a satisfied disinfectant effect,and be applied in clinic to meet the requests of high-level disinfection for gastrointestinal endoscopy.
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The quality control process throughout the Ebola virus nucleic acid detection in Sierra Leone-China Friendship Biological Safety Laboratory (SLE-CHN Biosafety Lab) was described in detail, in order to comprehensively display the scientific, rigorous, accurate and efficient practice in detection of Ebola virus of first batch detection team in SLE-CHN Biosafety Lab. Firstly, the key points of laboratory quality control system was described, including the managements and organizing, quality control documents and information management, instrument, reagents and supplies, assessment, facilities design and space allocation, laboratory maintenance and biosecurity. Secondly, the application of quality control methods in the whole process of the Ebola virus detection, including before the test, during the test and after the test, was analyzed. The excellent and professional laboratory staffs, the implementation of humanized management are the cornerstone of the success; High-level biological safety protection is the premise for effective quality control and completion of Ebola virus detection tasks. And professional logistics is prerequisite for launching the laboratory diagnosis of Ebola virus. The establishment and running of SLE-CHN Biosafety Lab has landmark significance for the friendship between Sierra Leone and China, and the lab becomes the most important base for Ebola virus laboratory testing in Sierra Leone.
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Humans , China , Ebolavirus , Classification , Genetics , Hemorrhagic Fever, Ebola , Diagnosis , Virology , Laboratories , Workforce , Reference Standards , Laboratory Infection , Quality Control , RNA, Viral , Genetics , Sierra LeoneABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Fuzhenghuayu compound (FZHc) on expression of nuclear factor E2-related factor 2 (Nrf2) in hepatocytes under conditions of hepatic fibrosis using a mouse model.</p><p><b>METHODS</b>Mice were randomly assigned to a control group and a hepatic fibrosis model group. The control group was further divided into three subgroups for use as normal controls (A1), mineral oil-treated controls (A2), and FZHc-treated controls (A3); the hepatic fibrosis model group was administered carbon tetrachloride (CC14 dissolved in mineral oil and injected intraperitoneally) and further divided into four subgroups for use as 6-weeks models (B1), 10-weeks models (B2), low-dose (L)-FZHc models (C1), and high-dose (H)-FZHc models (C2). The FZHc (capsule powder diluted with double-distilled water to 0.1 g/mL) was administered via gastric perfusion to groups A3, C1, and C2 starting at week 7 of the experiment. At the end of week 6 and 10, hepatic specimens were collected and evaluated for degree of hepatic fibrosis and inflammation using routine haematoxylin-eosin staining and Masson staining. Immunohistochemical analysis was performed to measure the hepatocyte expression of Nrf2, NAD(P)H quinine oxidoreductase 1 (Nqol), a-smooth muscle actin (a-SMA) and fibronectin (FN). Real-time fluorescence quantitative PCR was used to measure Nrf2 mRNA expression. Western blotting was used to detect Nrf2 and Nqol total protein expression and Nrf2 nuclear translocation. F test, LSD test and ridit test were used for statistical analyses.</p><p><b>RESULTS</b>Compared with the B2 group (ridit value: 0.09), the model groups treated with FZHc showed significantly lower degrees of hepatic inflammation and fibrosis for both the low (C1 group, ridit value: 0.32) and high doses (C2 group, ridit value: 0.40) (F =82.927, P less than 0.05). In addition, compared with the B2 group, the model groups treated with FZHc showed significantly decreased expression of a-SMA and FN proteins, with a dose-dependent trend (by immunohistochemistry: C 1 group at the end of 10 weeks, F =77.421, 118.262, P less than 0.05; C2 group, P =0.002, 0.013) and significantly increased expression of Nrf2 and Nqol proteins (by immunohistochemistry:C1 and C2 groups at the end of 10 weeks, F =182.537, 75.615, P less than 0.05 and by westen blotting: F =45.664, 127.673, P less than 0.05), which also showed a dose-dependent trend (C2 group, P =0.000, 0.014; 0.005, 0.014). Western blotting also indicated that the amount of nuclear transported Nrf2 was higher in the C1 and C2 groups at the end of 10 weeks (vs. B2 group, F =94.787, P less than 0.05), and the amount of nuclear transported Nrf2 was significantly higher in the C2 group (vs. C1 group, P =0.044). Nrf2 mRNA expression was significantly higher in the C1 group than in the B2 group (F =3230.105, P less than 0.05), and the C2 group had more substantially increased expression (P =0.001); there was no statistical difference found between groups B1 and B2 (P =0.094).</p><p><b>CONCLUSION</b>Fuzhenghuayu compound increased the expression of Nrf2 mRNA and protein under conditions of hepatic fibrosis in mice and stimulated Nrf2 nuclear transport, as well as increased expression of the Nrf2 target gene Nqol that is known to suppress activation of hepatic stellate cells and decrease the deposition of FN. Therefore, Fuzhenghuayu compound may ameliorate hepatocyte injury in hepatic fibrosis in mice by exerting an antihepatic fibrosis effect.</p>
Subject(s)
Animals , Female , Mice , Drugs, Chinese Herbal , Pharmacology , Hepatocytes , Metabolism , Liver Cirrhosis, Experimental , Metabolism , Mice, Inbred Strains , NAD(P)H Dehydrogenase (Quinone) , Metabolism , NF-E2-Related Factor 2 , MetabolismABSTRACT
Matrix protein 2(M2) is an integral tetrameric membrane protein of influenza A virus, which functions as ion channel. M2 sequence has shown remarkable conservation, so there has been growing interest in it as "universal" vaccine. In order to establish a stable 293 cell line that express M2 protein under the control of the tetracycline operator, M2 gene was obtained by PCR amplification from the plasmid containing the segment 7 of influenza A virus strain A/PR/8/34 firstly. The PCR product was cloned into BamH I/Not I restriction site of pcDNA5/FRT/TO vector, and cotransfected with pOG44 which express Flp recombinase into Flp-In T-REx-293 cell. Integration of pcDNA5/FRT/TO-M2 into the cell genome at the Flp Recombination Target (FRT) site brought the SV40 promoter and the initiation codon in frame with the hygromycin resistance gene. Thus, stable cell lines were selected for hygromycin resistance. The expression of M2 protein from hygromycin-resistant cell was induced by addition of tetracycline into the cell culture media, and then tested by indirect immunofluorescence assay (IFA). 16 strains with high expression of M2 were selected. After subculturing for more than ten passages, the cell lines still stably expressed M2 protein. No M2 protein could be detected without tetracycline induction, suggesting that the expression was strictly controlled by tetracycline operator. The cell lines expressing M2 will be useful for further functional studies of M2 protein, detection of immune response against natural structure M2 protein and development of live attenuated influenza virus vaccine with reverse genetics technique.
Subject(s)
Animals , Humans , Cloning, Molecular , Gene Expression , Genetic Vectors , Genetics , HEK293 Cells , Influenza A virus , Genetics , Metabolism , Influenza Vaccines , Genetics , Operator Regions, Genetic , Recombinant Proteins , Genetics , Tetracycline , Pharmacology , Transfection , Viral Matrix Proteins , GeneticsABSTRACT
In order to evaluate the response to vector-expressed M1 and HA genes of influenza virus in mice, we prepared recombinant plasmid pStar-M1/HA and recombinant adenovirus Ad-M1/HA containing both the full-length matrix protein 1(M1) and hemagglutinin (HA) genes of human H5N1 influenza virus strain A/Anhui/1/2005. We then combined the DNA vaccine and adenoviral vaccine in immunization of BALB/c mice with a prime-boost regime. We immunized the mice with DNA vaccine at day 0 and 28 and with recombinant adenoviral vaccines at day 14 and 42. We took blood samples before each injection and 14 days after the final injection for detection of humoral immune responses. At day 56, we sacrificed the mice and collected splenocytes for detection of cellular immune responses. ELISA and hemagglutination inhibition (HI) assay showed that specific IgG Abs against H5N1 influenza virus was induced in serum of the immunized mice. ELISPOT results confirmed that the specific cellular immune responses were successfully induced against the M1 and HA proteins of H5N1 influenza virus. This study provides new strategy for development of novel influenza vaccines.
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Animals , Mice , Adenoviridae , Genetics , Metabolism , Antibodies, Viral , Blood , Hemagglutinin Glycoproteins, Influenza Virus , Genetics , Allergy and Immunology , Immunization , Influenza A Virus, H5N1 Subtype , Allergy and Immunology , Influenza Vaccines , Allergy and Immunology , Mice, Inbred BALB C , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Vaccines, DNA , Allergy and Immunology , Viral Matrix Proteins , Genetics , Allergy and ImmunologyABSTRACT
We developed vectors expressing two antigen of H5N1 influenza virus. Based on the human H5N1 avian influenza virus strain A/Anhui/1/2005 isolated in China, we amplified the matrix protein 2 (M2) and Hemagglutinin (HA) genes by PCR and subcloned them into pStar vector to construct two genes co-expressing recombinant DNA vaccine pStar-M2/HA. After transfection of 293 cells with the plasmid, we confirmed with indirect immunofluorescence assay (IFA) that M2 and HA genes cloned on plasmid pStar co-expressed successfully. Using Ad-Easy adenovirus vector system, by homologous recombination in bacteria and packaging in 293 cells, we constructed two recombinant adenoviruses, namely Ad-M2 and Ad-HA. After infection of 293 cells with the recombinant adenoviruses, we confirmed with IFA that M2 and HA genes cloned into adenoviruses expressed successfully. We then combined the recombinant DNA vaccine and adenoviral vector vaccines in immunization of BALB/c mice with a prime-boost regime. On day 0 and day 28, we immunized the mice with DNA vaccine and on day 14 and day 42, with recombinant adenovirus vaccines. We took blood samples before each injection and 14 days after the final injection. On day 56, we collected splenocytes from the mice. ELISA and hemagglutination inhibition (HI) assay showed that the vaccines successfully induced specific IgG antibodies against HA protein in serum of the immunized mice. ELISPOT confirmed that the vaccines successfully induced the special cellular immune response to M2 and HA protein of H5N1 influenza virus. The study on combined immunization with M2 and HA genes provided basis for development of novel influenza vaccine.
Subject(s)
Animals , Female , Mice , Adenoviridae , Genetics , Metabolism , Genetic Vectors , Genetics , Hemagglutinin Glycoproteins, Influenza Virus , Genetics , Influenza A Virus, H5N1 Subtype , Genetics , Allergy and Immunology , Influenza Vaccines , Allergy and Immunology , Mice, Inbred BALB C , Recombinant Proteins , Genetics , Allergy and Immunology , Vaccination , Vaccines, DNA , Allergy and Immunology , Viral Matrix Proteins , GeneticsABSTRACT
Virus-like particles (VLPs) structurally mimic the authentic virus whereas contain no viral genome. VLPs technique plays an important role in basic research such as virus assembly and virus morphology diversity. With special immunology properties, VLPs vaccine can induce immune response effectively. VLPs can act as adjuvant by regulating dendritic cells. Other adjuvant or polypeptide can be integrated into VLPs to construct chimeric vaccines. With the capability of packaging nucleic acid or other small molecules, VLPs can be used for vehicles to deliver these substances under suitable conditions. VLPs can substitute natural virus in immunology assay.