OBJECTIVETo establish an HPLC method for the simultaneous determination of three major bioactive components in Jingzhi Guizhi Fuling capsules namely laetrile, paeoniflorin and paeonol.

METHODA LiChrospher C18 column (4.6 mm x 250 mm, 5 microm) was used. The chromatography was carried out with a stepwise gradient programming. The mobile phase was acetonitrile-water (containing 0.1% phosphorous acid) and the flow rate was 1.0 mL x min.

RESULTThe linear range of laetrile was 12.87-102.94 micron x mL(-1), r = 0.999 9, paeoniflorin 24.84 - 198.7 microg x mL(-1), r = 0.9999 and paeonol 12.57-100.56 microg x mL(-1), r = 0.999 9. The method is accurate with variation less than 1.5 % and recovery more than 95 %.

CONCLUSIONThe method was successfully applied to analyze three major bioactive components in Jingzhi Guizhi Fuling capsules.

Acetophenones ; analysis ; Amygdalin ; analysis ; Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; Capsules ; Chromatography, High Pressure Liquid ; methods ; Cinnamomum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Glucosides ; analysis ; Monoterpenes ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Polyporales ; chemistry ; Reproducibility of Results

Objective To study the expressions of vascular endothelial growth factor (VEGF) and Ki-67 antigen and their correlation in human brain astrocytorna. Methods Immunohistochemistry with SP method was employed to detect the expressions of VEGF and Ki-67 in 60 human brain astrocytoma tissue (grade Ⅰ-Ⅱ 28 cases, grade Ⅲ 20 cases, grade Ⅳ 12 cases) and 30 normal tissues adjacent to the tumors. Results VEGF and Ki-67 were not expressed in the normal brain tissues adjacent to the astrocytoma, but their expressions were detected in all the astroeytoma tissues of different grades (P<0.05). High-grade astrocytomas showed significantly higher VEGF and Ki-67 expression than low-grade astrocytomas (P<0.05), and a positive correlation was noted between VEGF and Ki-67 expressions (r=0.310, P<0.05). Conclusion VEGF and Ki-67 expressions can provide important evidence for evaluating the malignancy and clinicopathologieal grading of human astrocytoma.

Objective To understand the distribution and drug resistance of pathogenic bacteria from diabetic foot patients of different grades of Wagner classification,and provide basis for the rational use of antimicrobial agents in clinical practice.Methods Clinical data,pathogens isolated from wound secretion,and antimicrobial susceptibility testing result of diabetic foot patients who were admitted in the department of endocrinology in a hospital between January 2014 and December 2016 were analyzed retrospectively,patients were divided into gram-positive (G+) bacteria infection group,gram-negative (G) infection group,and mixed infection group according to the isolation of bacterial species,relationship between patients' clinical characteristics and Wagner grades was analyzed.Results There were 103 patients with diabetic foot,39 with G+ bacterial infection,50 with G-bacterial infection,and 14 with mixed infection.The age and disease course of patients with mixed infection was significantly elder and longer than the other two groups of patients(both P＜0.05);glycosylated hemoglobin (HbA1c) and procalciton in mixed infection group were both lower than the other two groups(both P＜0.05);the percentage of neutrophils in G+ infection group was higher than the other two groups(P＜0.01).A total of 132 specimens were detected,103 strains were isolated,the positive rate was 78.03％,isolation rates of G+ and G-bacteria were 33.33％ (44/132) and 44.70％ (59/132) respectively.The major G+ strains were Staphylococcus aureus (n =19) and Staphylococcus epidermidis(n=11),the major G bacteria were Citrobacterbraakii(n=12),Escherichia coli(n=11),and Pseudo monas aeruginosa (n =11).As the Wagner grade increased,the proportion of mixed infection continued to rise.The main multidrug-resistant organisms were methicillin-resistant Staphylococcus aureus and extended-spectrum betalactamase-producing G-strains.Conclusion For diabetic foot patients,especially those with higher Wagner grades,etiological examination and antimicrobal susceptibility testing of ulcer secretion should be timely performed,so as to provide reference for rational use of antimicrobial agents in clinic.

Trichosporon species now ranks as the second most common cause of disseminated yeast infections with a high mortality rate. Breakthrough trichosporonosis in patients receiving echinocandins therapy is being recognized recently. We present a case of breakthrough trichosporonosis with acute viral myocarditis while receiving caspofungin therapy. Trichosporon infection should be considered in patients, who have risk factors for invasive fungal infection and develop unexplained clinical manifestations of infection despite treatment with echinocandins.
Adult ; Antifungal Agents ; therapeutic use ; Echinocandins ; therapeutic use ; Female ; Humans ; Lipopeptides ; Treatment Outcome ; Trichosporonosis ; drug therapy ; microbiology

Objective To observe the risk factors and prevalence status of hypertension at the Guangxi Zhuang-rural region.Methods Through cross-sectional survey,Xinlan village (Ethnic-Zhuang tribe village) in Liangqing District of Nanning was chosen as survey site.Select Zhuang-ethnic villagers (age≥18 years,living time ≥ 6 months per year) as survey subjects.Questionnaire survey and examinations including blood pressure measurement,blood tests,and urine test were made by trained medical professionals.Multivariate logistic regression analysis was used to analyze the risk factors of hypertension.Results Complete data were obtained in 2036 residents.The hypertension prevalence rate was 11.6％ (237/2036) [12.5％ (110/883) for male,l1.0％ (127/1153) for female],adjusted prevalence rate was 10.7％ (11.3％ for male,10.4％ for female) based on the population of Guangxi in 2010.Hypertension awareness rate was 36.3％ (86/237),therapy rate was 22.8％ (54/237),and control rate was 11.0％ (26/237).Multivariate logistic regression analysis showed that age (OR =1.06,95％ CI:1.05-1.08,P ＜ 0.01),alcohol drinking habit (OR =1.92,95％ CI:1.25-2.95,P ＜ 0.01),high salt diet(OR =1.48,95％ CI:1.04-2.10,P ＜ 0.05),diabetes (OR =7.03,95％ CI:1.94-25.44,P ＜0.01),obesity (OR =1.20,95 ％ CI:1.15-1.25,P ＜ 0.01) and hypertriglyceridemia (OR =1.12,95％ CI:1.03-1.22,P ＜0.01) were risk factors for hypertension in this cohort.Conclusions Prevalence rate is slightly lower in adult of the Guangxi Zhuang-rural areas than national or Guangxi Zhuang Autonomous Region levels,but hypertension awareness,therapy and control rates in this cohort are not satisfactory.Targeted strategies are needed to improve the prevention and therapy of hypertension in this region.

OBJECTIVETo study the alterations of follicular T helper cells (CD4(+)CXCR5(+)Tfh cells, Tfh) on circulating T lymphocytes in children with asthma, and to study the expression of transcription regulatory factors BCL-6 and BLIMP-1 mRNA.

METHODSSixty-four children with asthma and 25 healthy controls were enrolled in this study. On the basis of the disease, the children with asthma were classified into acute phase group (n=36) and remission phase group (n=28). The flow cytometry was used to detect the proportion of CD4(+)CXCR5(+)Tfh cells on CD4(+)T lymphocytes. Real-time PCR was performed to detect the levels of BCL-6 mRNA and BLIMP-1 mRNA. The double -antibody Sandwich ELISA was used to detect plasma concentrations of total IgE, IL-2, IL-6 and IL-21.

RESULTSThe proportion of CD4(+)CXCR5(+)Tfh cells was significantly higher in the acute group than in the control group and the remission group (P<0.05). Transcription levels of BCL-6 mRNA were significantly higher, while the inhibitory factors BLIMP-1 mRNA was significantly lower in the acute group than in the remission group and control group (P<0.05). The plasma concentration of IL-6 in the acute group increased significantly compared with the control group (P<0.05). Plasma concentrations of total IgE and IL-21 increased significantly, in contrast, plasma IL-2 concentration decreased significantly in the acute group, compared with the control group and the remission group (P<0.05). Correlation analysis showed that both IL-21 and IL-6 concentrations were positively correlated with the proportion of CD4(+)CXCR5(+)Tfh cells (r=0.76, r=0.46 respectively; P<0.05), while IL-2 level was negatively correlated with the proportion of Tfh cells (r=-0.68, P<0.05).

CONCLUSIONSThe abnormal proportion of CD4(+)CXCR5(+)Tfh cells might be involved in the immunological pathogenesis of acute asthma in children. The increased expression of BCL-6 mRNA and decreased expression of BLIMP-1 mRNA as well as the alterations of plasma total IgE, cytokines IL-2, IL-6 and IL-21 in microenvironment might be account for the increased proportion of CD4(+)CXCR5(+)Tfh cells in children with acute asthma.

Asthma ; immunology ; Child ; Child, Preschool ; DNA-Binding Proteins ; genetics ; Female ; Humans ; Immunoglobulin E ; blood ; Infant ; Interleukins ; blood ; Male ; Positive Regulatory Domain I-Binding Factor 1 ; Proto-Oncogene Proteins c-bcl-6 ; RNA, Messenger ; analysis ; Receptors, CXCR5 ; analysis ; Repressor Proteins ; genetics ; T-Lymphocytes, Helper-Inducer ; immunology

Objective To study the genotypic drug-resistant mutation among treat-naive or treated patients infected with HIV-1 CRF01_AE in Zhejiang province during 2004-2007. Methods HIV-i pol amplicons (PR+RT) from 13 treated and 43 treat-naive patients were obtained by reverse transcription-polymerase chain reaction (RT-PCR). The sequences were analyzed for genotypic antiretroviral resistance through online tools (http://hivdb.stanford.edu). Results The median count of CD44+ T lymphocytes in 43treat-naive patients was 229 cells/mm3 and the median log10 viral load was 3.41. Some drug-resistant mutations were seen in these samples including amino acid 10, 46, 71, in the genes of protease (PR) and 103, 118, in the genes of reverse transcriptase (RT) whereas twenty-nine resistance mutations in the genes of PR and RT were obtained in the 13 treated patients (8/13, 61.5% ). The high prevalence of drug-resistant mutations was observed in patients who had been receiving HAART (hight active antiretroviral therapy). Among them, cross drug resistance was dominant. Correspondingly, the median counts of CD44+ T lymphocytes and the log10 viral load were 186 cells/mm3 and 3.91. Conclusion There was a low prevalence of genotypic drug-resistant mutations in treat-naive patients, but higher drug-resistant mutation in treated patients. More attention should be paid to the transmission of drug-resistant HIV strains and the antiretroviral therapy recipe should be adjusted correspondingly for the development of ART drugs, intervention as well as clinical therapy programs.

Objective To examine the prevalence and correlates for needle-sharing among new and long-term injection drug users (IDUs) in Sichuan province, China. Methods Three consecutive cross-sectional surveys were conducted in 6 prefectures of Sichuan province, from 2003 to 2005.Questionnaire-based interviews provided information including socio-demographics, drug-use, sexual behaviors, and HIV-related services. Multivariable logistic regression methodology was used for data analysis. Results Of 3852 eligible participants, the rates of needle-sharing in the last 6 months for IDUs with a less than one year, one to three year, and more than three year injection history were 19.9%,29.1%, and 36.3%, respectively. Needle-sharing for IDUs with a less than one-year injection history was independently associated with factors as: being female, having minority background, higher frequency of injection, sharing injecting equipments, ever having had a non-regular sex partner, or a regular sex partner who injected drugs, never had a test on HIV, being recruited in 2005, and from Zigong prefectm'es of Sichuan. Needle-sharing for IDUs with a 1-3 year injection history were independently associated with receiving less education, having higher frequency of injecting behavior, sharing injecting equipments,ever having had a non-regular sex partner/a regular sex partner who injected drugs, ever having had unprotected sex with a non-regular sex partner, ever having had unprotected sex with a commercial sex partner or client, never attended skill training for HIV/STD prevention, being recruited in 2005 and 2004,and being recruited from Deyang, Zigong, and Liangshan prefectures. Needle-sharing for IDUs with a more than three-year injection history was independently associated with factors as: less education, higher frequency of injecting, sharing injecting equipment, ever had a non-regular sex partner, ever had unprotected sex with a non-regular sex partner, ever had unprotected sex with a commercial sex partner or client, having had regular sex partners who inject drugs, never attended needle exchange programs, being recruited in 2005 and 2004, and being recruited from Deyang, Zigong, and Liangshan prefectures. Conclusion The rates for both needle/paraphernalia sharing and unprotected sex behaviors were high in the IDUs in Sichuan province. These rates were higher among longer-term IDUs than in new IDUs. Our data underlined that better targeted, in-depth and sustained comprehensive intervention packages which including needle exchange, condom promotion along with distribution, and education, were urgently needed to reduce both IDU-related and sex-related risk behaviors among both new and longer term IDUs.

ObjectiveTo observe the morphological changes of bone in the progress of chronic fluorosis.MethodsWistar rats were randomly divided into three groups,30 rats in each group:normal control group,experimental group Ⅰ and experimental group Ⅱ according to body weight.Rats in normal control group drank distilled water freely.Experimental group Ⅰ and group Ⅱ drunk distilled water with sodium fluoride preparation of fluorine containing ion 100,150 mg/L solution for six months,respectively.Bone mineral density was detected by X-ray,bone morphological changes were observed under light microscope and bone histomorphometric parameters were calculated using image analysis software.ResultsThe bone mineral density values were different statistically between the three groups after feeding for 2 and 4 months(F =19.79,3.28,all P ＜ 0.05).However no significant difference was found after feeding for 6 months(F =1.80,P ＞ 0.05).The bone mineral density of experimental group Ⅰ (0.20 ± 0.03,0.21 ± 0.03) was significantly higher than that of the normal control group(0.17 ± 0.03,0.20 ± 0.04) after feeding for 2 and 4 months.The bone mineral density of experimental group Ⅱ (0.21 ± 0.02) was lower than that of normal control group(0.22 ± 0.03) after feeding for 6 months.The bone lamella in experimental group Ⅰ was arranged disorderly,the number of osteocytes increased with their nucleus atrophy and the osteoblasts were more than that of control grouo which arranged in layers observed under light microscooy.In exoerimental group Ⅱ,the bone lamella was bent deformation,the number of osteocytes had decreased with their nucleus shrinking or even disappeared and the number of osteoclasts had increased significantly observed under light microscopy.In experimental group Ⅰ,the mean trabecular density [(0.33 ± 0.03)％] increased and the mean trabecular separation,thickness [( 163.57 ± 1.99),(59.26 ± 7.18 ) μm] decreased compared with that of normal control group [(0.31 ± 0.02)％,(186.60 ± 2.90)μm,(86.42 ± 1.48)μm,all P ＜ 0.05].In experimental group Ⅱ,the mean trabecular density[(0.26 ± 0.02)％] decreased,the mean trabecular thickness[(71.42 ± 10.77)μm] reduced compared with that of normal control group[(0.31 ± 0.02)％,(86.42 ± 1.48)μm].ConclusionsExcess fluoride can damage bone tissue.Low doses of fluoride can stimulate osteoblast activity and enhance osteogenesis.The activity of osteoblasts is great than that of osteoclasts.High doses of fluoride can stimulate both osteoblasts and osteoclasts activity,but mainly the activity of osteoclasts,and bone resorption increases.

OBJECTIVETo determine whether 3'phosphorothioate-modified-2 terminal mismatched primers can turn off DNA polymerization mediated by Exo(+) polymerase.

METHODSTwo-directional primer extension was performed using polymerase with and without 3' exonuclease activity. The effects of unmodified primers and 3' phosphorothioate-modified primers on primer extension were evaluated.

RESULTSExo(-) polymerase yielded products from matched and mismatched primers regardless of their modification. However, 3' phosphorothioate-modified primers with a single base mismatch at -2 position worked similarly to the terminal (-1) mismatched primers in triggering the novelly reported "off-switch" of Exo(+) polymerase.

CONCLUSIONThese data suggested that the "off-switch" can be of enormous application in the diagnosis of single gene diseases and in the association studies by single nucleotide polymorphism screening.

DNA Primers ; chemistry ; genetics ; Exonucleases ; metabolism ; Humans ; Phosphorothioate Oligonucleotides ; chemistry ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide