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OBJECTIVE：To identify Amomum villosum from different habitats and its adulterants. METHODS ：Through the identification methods of microscopic characteristics ，microscopic characteristics maps of 9 batches of A. villosum from genuine producing areas ，domestic commercially available A. villosum and its adulterants were obtained. The feature maps were extracted digitally and analyzed by SPSS 21.0 software. RESULTS ：Commercially available A. villosum was mainly from Guangdong ， Guangxi，Yunnan and Fujian ；the collected adulterants of A. villosum included A. villosum Lour. var. xanthioides T.L.Wu et Senjen ， A. aurantiacum H. T. Tsai et S. W. Zhao and other A. species from Yunnan Xishuangbanna ， Laos and Myanmar. Under the microscope，it was observed that microscopic characteristics of surface （such as exocarp color ，prickle，non-glandular hairs ， endocarp color ，endocarp oil chamber ） of A. villosum from different habitats and its adulterants were different. There was statistically significant difference in fruit width values and endocarp oil point diameter among all samples （P＜0.05）. CONCLUSIONS：The microscopic characteristics maps of A. villosum from different habitats and its adulterants by the microscopic characteristics identification methods will make up for the deficiency of traditional experience identification. The quantitative analysis of micro-property and the establishment of micro-property database of A. villosum can provide reference for the property identification and quality control of this medicinal material.
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Objective:To investigate the clinical characteristics and pathogen spectrum of acquired immunodeficiency syndrome (AIDS) patients complicated with pulmonary filamentous fungal infection in Guangdong Province, so as to provide evidences for improving the diagnosis and treatment.Methods:A total of 143 AIDS patients with pulmonary filamentous fungal infection hospitalized in Guangzhou Eighth People′s Hospital, Guangzhou Medical University from January 2016 to December 2018 were included. The filamentous fungi cultured in bronchoalveolar lavage fluid of these patients were identified with morphological and molecular biological methods. And their clinical characteristics were analyzed. Nonparametric Kruskal-Wallis H test and chi-square test were used for statistical analysis. Results:Among the 143 patients, 116(81.1%) had fever, 104(72.7%) had cough, 83(58.0%) had expectoration, and 59(41.3%) had anhelation. The CD4 + T lymphocyte count was 22.0(9.3, 60.8) cells/μL and 118(82.5%) cases were below 100.0 cells/μL. The white blood cell counts decreased in 52(36.4%) cases and increased in 18(12.6%) cases, anemia was found in 109(76.2%) cases, platelet count decreased in 29(20.3%) cases. Sixty-four (44.8%) cases were positive for galactomannan test. Chest computed tomography showed diffuse infection of both lungs in 114(79.7%) cases, miliary changes in 12(8.4%) cases, pleural effusion in 44(30.8%) cases, and enlargement of pleural and (or) mediastinal lymph nodes in 45(31.5%) cases. After receiving antifungal therapy, 124 (86.7%) cases were cured or improved, and 19 (13.3%) cases were discharged automatically or died of disease deterioration. Among the 143 strains of filamentous fungi, there were 56 strains of Aspergillus species pluralis (39.2%, including 24 strains of Aspergillus fumigatus), 37 strains of Talaromyces marneffei ( T. marneffei) (25.9%), 22 strains of Penicilium species pluralis (15.4%), and 28 strains of other genera of filamentous fungi (19.6%). The median CD4 + T lymphocyte counts in patients infected with Aspergillus species pluralis, T. marneffei, Penicilium species pluralis and other genera were 24.5, 15.0, 53.5 and 22.0 cells/μL, respectively, and the difference was statistically significant ( H=11.282, P=0.010). The proportions of AIDS patients with different pulmonary filamentous fungal infection of CD4 + T lymphocyte count ≤50.0 cells/μL in descending order were T. marneffei group (89.2%(33/37)), Aspergillus species pluralis group and other genera group (67.9%(38/56), 67.9%(19/28)), and Penicillium species pluralis group (54.5%(12/22)), and the difference was statistically significant ( χ2=9.296, P=0.026). Conclusions:The clinical manifestations of pulmonary filamentous fungal infection in AIDS patients in Guangdong Province are not specific. The pathogenic spectrum contains various genera, and T. marneffei and Aspergillus fumigatus are dominant, which could be correlated with CD4 + T lymphocyte count.
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Objective To explore the pathogen spectrum, drug resistance rate and clinical characteristics of pneumonia caused by non-tuberculous mycobacteria (NTM) in acquined immuno-deficiency syndrome (AIDS) patients.Methods The clinical data of 31 hospitalized AIDS patients with bronchoalceolar lavage flind (BALF) culture confirmed NTM pulmonary disease in Guangzhou No.8 People′s Hospital from January,2008 to February,2015 were retrospectively analyzed, including pathogen spectrum, drug resistance rate and clinical characteristics.The clinical characteristics and drug resistance were compared between Mycobacterium avmm-intracellulare complex (MAC) pneumonia and the non-MAC pneumonia, and t test and chi-square test were used.Results Of the 31 AIDS patients,28 were male and 3 were female, with the mean age of 40.9 years old.The 31 NTM strains were consisted of 14 MAC strains and 17 non-MAC strains (including 4 M.kansasii strains,3 M.lentiflavumstrains, 2 M.szulgai strains, 2 M.yongonense strains etc).There was no significant difference between two groups in sex ratio, mean age, clinical manifestations, laboratory tests and treatment outcome (all P>0.05).The major clinical manifestations included fever, productive cough, weight loss, anemia and low CD4+ count (<50/μL).Most patients showed thoracic lymphadenectasis and patchy shadows in lungs, and few patients had millet shadows and pericardial effusion.Compared with non-MAC strains, MAC strains had higher drug resistant rate of moxifloxacin (10/14 vs 4/17), levofloxacin (14/14 vs 8/17), and clarithromycin (11/14 vs 7/17).More extensively drug resistance strains were seen in non-MAC strains compared with MAC strains (11/14 vs 7/17).Conclusions MAC is the most common pathogen of NTM pulmonary disease in AIDS patients.The clinical features of pneumonia caused by MAC and non-MAC are similar, but drug resistance of MAC strains are more severe.
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Objective To explore the diagnostic value of Talaromyces marneffei (T.marneffei)-specific mannose glycoprotein Mp1p antigen for T.marneffei infection in acquired immune deficiency syndrome (AIDS) patients.Methods All cases were recruited in this study from January 2012 to June 2015 in Guangzhou No.8 People′s Hospital, including 184 AIDS patients with T.marneffei infection confirmatively diagnosed by culture, and 205 controls including 176 AIDS patients without T.marneffei infection and 29 health controls.Double antibody sandwich enzyme linked immunosorbent assay and fluoroimmunoassay combined with double-antibody sandwich were both utilized to detect serum Mp1p antigen levels, and their sensitivity and specificity for diagnosing T.marneffei infection in patients with AIDS were analyzed.x2 test and t test were used for statistical analysis.Results The ratio of males to females and age of the study group were both comparable to those of the control group (x2=0.019, P=0.889;t=1.810,P=0.07, respecitvley).The sensitivities of double antibody sandwich enzyme linked immunosorbent assay and fluoroimmunoassay combined with double-antibody sandwich were 82.07%(151/184) and 83.15%(153/184), respectively (x2=0.076, P=0.783).The specificities were 93.17%(191/205) and 92.68%(190/205), respectively (x2=0.037, P=0.847).The accuracy values were 87.92%(342/389) and 88.17%(343/389), respectively (x2=0.012, P=0.912).The false positive rates were 6.83%(14/205) and 7.32%(15/205), respectively.The false negative rates were 17.93%(33/184) and 16.85%(31/184), respectively (x2=0.049, P=0.829).The positive predictive values were 91.52%(151/165) and 91.07%(153/168), respectively (x2=0.021, P=0.886).The negative predictive values were 85.27%(191/224) and 85.97%(190/221), respectively (x2=0.045, P=0.832).The Kappa values were 0.83 and 0.80, respectively.Conclusion Detection of serum Mp1p antigen of T.marneffei possesses high specificity and sensitivity, which may be utilized for rapid and early diagnosis of T.marneffei infection in patients with AIDS.
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OBJECTIVE:To establish a method for the content determination of glabridin in Clycyrrhizae Radix et Rhizoma from different regions. METHODS:HPLC was performed on the column of Inertsil ODS-SP with mobile phase of acetoni-trile-0.05%phosphoric acid(gradient elution)at a flow rate of 0.8 ml/min,detection wavelength was 280 nm,the column tempera-ture was 35 ℃,and the injection volume was 20 μl. RESULTS:The linear range of glabridin was 0.906-18.12 μg/ml(r=0.999 7), RSDs of precision,stability and reproducibility tests were lower than 3%;recovery was 98.73%-101.90%(RSD=1.25%,n=6). There were obvious differences among the glabridin contents in G. uralensis from different regions. CONCLUSIONS:The method is simple and accurate with high precision,and can be used for the content determination of glabridin in Clycyrrhizae Radix et Rhi-zoma.
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Objective To investigate gene polymorphisms of drug targets and mutations associated with drug resistance in Pneumocystis jiroveci (P.jiroveci) isolates.Methods Among 148 samples isolated from human immunodeficiency virus (HIV)infected patients with pneumonia in Guangdong,mitochondrid larg subunit rRNA (mtLSUrRNA) gene was amplified from 51 samples.Dihydropteroate synthase (DHPS),dihydrofolate reductase (DHFR) and Cytochrome b (CYB) genes of P.jiroveci were detected by gene sequencing,and compared with the reference sequences in GenBank to evaluate gene polymorphisms.Results P.jirovecii DHPS,DHFR and CYB genes were all successfully amplified from 51 samples.For DHPS gene,48 (94.1％) were wild-type and 3 (5.9％) had gene mutation associated with drug resistance.For DHFR gene,30 were wild-type,and 21 had a synonymous mutation at position 312,and 1 nonsynonymous mutation at position 188.There were no mutations associated with drug resistance.For CYB gene,polymorphisms of were detected at 5 sites,4 of which were synonymous mutations,1 was non-synonymous mutation.No mutation associated with drug resistance was found.Based on the gene polymorphism of CYB6,the strains can be classified into 6 genotypes,and 2 were first detected,including 25 CYB1,13 CYB2,2 CYB5,4 CYB8,as well as newly detected 4 CYB10 and 3 CYB11 strains.Conclusions The mutations associated with drug resistance in P.jiroveci isolates in Guangdong remain uncommon.CYB gene shows gene polymorphisms and can be selected as one of targeted genes for multilocus sequence typing.
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Objective To investigate the characteristics of mycobacteria species distribution in human immunodeficiency virus (HIV)-positive patients co-infected with mycobacteria in Guangzhou. Methods A total of 133 mycobacteria strains isolated from HIV-positive patients and 150 strains isolated from HIV-negative patients were included in this study. After DNA extraction of mycobacteria, mycobacteria species identification was performed by sequencing of multiple genes.Differences in the identified species were compared between patients with and without HIV infection and the correlation between CD4 + T cells level and the mycobacterial species distribution was analyzed.Chi-square test was used for statistical analysis.Results Of the 133 mycobacteria strains isolated from HIV-positive patients, 82 were identified as Mycobacterium tuberculosis complex (MTC ). Fifty-one were identified as nontuberculous mycobacteria (NTM),of which the main species was Mycobacterium avium complex (MAC,31/51).Of the 150 mycobacteria strains isolated from HIV-negative patients,126 were identified as MTC and 24 as NTM,of which the main species was Mycobacterium abscessus (9/24).In patients with CD4 + T cell counts ≤100/μL,the positive rate of mycobacteria was 75 .94%(101/133),93.55 %(29/31) of MAC and 85 .00%(17/20)of other NTM.When the CD4 + T cell counts >100/μL,the positive rate for mycobacteria were all obviously decreased.Conclusions The proportion of NTM infection is higher in HIV-positive patients than HIV-negative patients in Guangzhou. Among HIV-positive patients > the most prevalent NTM species is MAC, while Mycobacterium abscessus is the most common species in HIVnegative patients. Mycobacterial infection in acquired immunodeficiency syndrome patients is closely associated with low CD4+ cells level.
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Objective To observe the effect of acupoint magnetic therapy on coronary heart disease.Methods 80 patients with coronary heart disease were divided into treatment group( n =40) and control group( n =40).The treatment group was treated with acupoint magnetic therapy on the basis of conventional treatment,and the control group with conventional treatment.Clinical therapeutic effect,the scores of traditional Chinese medicine symptom were observed.Results There was significant difference between the two groups in the total effective rate and the scores of traditional Chinese medicine symptom[90.0％ vs 72.5％,(13.25 ± 3.68 ) vs (15.18 ± 4.16),all P ＜0.05].Conclusion Acupoint magnetic therapy for treatment of coronary heart disease has an obvious therapeutic effect.
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Objective To detect dengue virus infection by serological method and to determine the sequences of E gene of dengue virus isolated from Guangzhou in 2006.so as to clarify the possible origin of dengue fever.Methods IgM and IgG antibodies to dengue virus were detected by immunochromatographic test(ICT);NSI antigen and IgM antibody were detected by enzyme-linked immunosorbent assay(ELISA).The virus was cultured and isolated from the serum samples within 2 days using C6/36 cell lines and was identified by immuno-fluorescence assay(IFA)and RT-PCR.The E gene of isolated virus DV1-GZ42/06 was sequenced;homological analysis and phylogenetic tree analysis were performed by comparing with the reference strains and epidemic virus strains.Results The positive rates of IgM and IgG of dengue virus in patients were 89.5%(433/484)and 38.0%(184/484)by ICT,respectively.The positive rates of NS1 antigen were 92.7%(38/41)in day 1 to day 2,83.3%(70/84)in day 3 to day 5,and 10.9%(5/46)in day 6 to day 10;and the IgM detection rates were 2.4%(1/41),51.2%(43/84)and 97.8%(45/46)at the same period by ELISA.Twenty-five strains of dengue virus were isolated from 41 serum samples(6 1.O%)and were identified as type 1 dengue virus by IFA and RT-PCR.The sequencing and phylogenetic analysis of the E gene showed that the homology between the isolated Guangzhou/42/06 strain and standard strain Hawaii/45 was 94.6%.and it had a high homology with the Thailand/NI09V104,Vietnam/06.and Vietnam/07 isolates(99.0%,98.6%and 98.6%,respectively)and belonged to the same cladogram,but had low homology with the isolated strain from Guangdong before 2006.Conclusions The detection of NS1 antigen is important in the early diagnosis of dengue fever.The outbreak of dengue fever in Guangzhou in 2006 was possibly caused by the cases from neighboring countries.
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OBJECTIVE To find out the distribution and durg resistance of the pathogens in 2008 in the 8th People′s Hospital of Guangzhou.METHODS Disc diffusion test(KB methods) was employed to study the antimicrobial susceptibility.WHONET 5.3 was applied for date analysis.RESULTS 208 Gram positive coccus strains accounted for(46.3%),241 Gram negative bacillus strains(53.67%) in A total of 449 pathogenic strains.Meticillin resistant strains accounted for 41.7%and Staphylococcus aureus and CNS for 55%.All the Staphylococcus strains were not Vancomycin-resistance.Escherichia coli and Klebsiella pnenmoniae strains producing extended spectrum-beta-lactamas accounted for 41.4%and 44.2%,but were highly sensitive to Carbapenem.Acinetobacter baumannii and Pseudomonas aeruginosa were highly sensitive to Carbapenem.CONCLUSIONS The resistance of antibiotics should be continuously monitored to provide basis for reasonable use of drugs in clinical practice and take effective evidence for the control of the problem.
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Objective To investigate in vitro activities of 5 fungal agents(amphotericin B,ketoconazole,fluconasole,5-fluorocytoaine and itroconazole)against Penicillium marneffei,providing references for clinical treatment.Methods E-test was used to test the in vitro susceptibilities of 5 antifungal agents (amphotericin B,ketoconazole,fluconasole,5-fluorocytoaine and itroconazole)against yeast form and mycelial form of 52 Penicillium marneffei strains isolated from our hospital.Results The 90% minimal inhibitory concentrations(MIC90)of amphotericin B,ketoconazole,fluconasole,5-fluorocytoaine and itroconazole against yeast form of Penicillium marneffei were 0.250,0.160,24.000,4.000,0.006 mg/L,the minimal inhibitory concentration(MIC)ranges were 0.004-0.500,0.002-0.016,1.000-256.000,0.002-32.000 and 0.002-0.008 mg/L,respectively;the MIG90 of the 5 agents against mycelial form of Penicil lium marneffei were 1.500,0.125,256.000,24.000 and 0.012 mg/L,respectively;the MIC ranges were 0.064-4.000,0-006-0.940,1-000-256.000,0.125-32.000 and 0.002-0.064 mg/L respectively.Five antifungal agents had different susceptibility patterns against both forms of Penicillium marneffei.And itroconazole was the most susceptible one,ketoconazole follows as the second.There was significant difference between the MICs of the same agent against yeast form of Penicillium marneffei and mycelial form of Penicillium marneffei.Conclusion In vitro antifungal agents suscepbibility tests of Penicillium marneffei can provide important references information for clinical treatment.
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Objective To evaluate bone marrow smear examination in early diagnosis of AIDS complicated with disseminated Penicillium marneffei infection. Methods Seventy-three clinically suspected AIDS patients complicated with disseminated PeniciUium marneffei infection were included in the study. Peripheral blood and bone marrow smear examinations, and the fungal thermally dimorphic culture were performed in all cases. Results PeniciUium marneffei infection was identified in 44 patients by peripheral blood and bone marrow fungal thermally dimorphic culture. The features of the bone marrow smear were as follows : they were all hyperplastic or significantly hyperplastic; there were thickened and increased granules, vacuolization and band-formed in most granulocytes; there were increased and augmented histiocytes, and increased plasma cells. In 12 samples of bone marrow smear, there were phagncytized mulberry-like Penicillium marneffei organisms in the cytoplasm of the histiocytes or the organisms found extracellularly. One sample demonstrated the increased granulocytes and the phagocytized organisms in the neutrophils and monocytes. In 4 samples of peripheral blood smear, there were phagocytized Penicillium marneffe organisms in the neutrophils and monocytes. Conclusion Bone marrow smear examination is of value in early diagnosis of AIDS complicated with disseminated Penicillium marneffei infection, which is 7 to 10 days earlier than routine fungal thermally dimorphic culture.
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Objective To study the pathogens and drug resistance profiles of pulmonary infection in patients with AIDS. Methods The pathogens and their drug susceptibility of pulmonary infection diagnosed by fibrobronchescopy-induced brunchoalveolar lavage fluid (BAI.F) culture and/or transbronchial biopsy in 116 AIDS cases were analyzed. Results Monopathogenic infection in lungs were detected in 18 cases(15.5%) and mixed infection in 98 cases ( 84.5%). Of the 116 cases, bacteria were present in 91 patients, fungi in 62, tubercle bacillus in 49, pneumocystis jiroveci in 29, and cytomegalovirus in 11.Ninety-five bacterial strains were isolated from BALF, mainly including Streptococci (34), coagulase negative Staphylococcus (20), Klebsiella pneumoniae (10) and Escherichia (7). The isolated bacteria were resistant to β-lactam, macrolides, quinolones and aminoglycosides, of which were 14 methicillin-resistant Streptococci (MRS) strains and 12 extended spectrum β-lactamases (ESBL) strains. Sixty-eight fungal strains were isolated, including 36 Candida mycodermas, 19 Penicilliums, 6 Aspergilli and 5 Mold fungi;they were sensitive to amphotericin B but resistant to fluconazol (5.6% -50. 0% ) and itraconazole( 10. 5%-60. 0% ). Conclusion Pneumonia in AIDS patients are usually caused by multiple pathogens,predominantly consisting of multiresistant bacteria and fungi. Therefore, antibiotics should be rationally chosen according to drug susceptibility test.