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.

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 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.

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

To characterize the complete genome sequence of coxsackievirus B1 (CVB1) MSH/KM9/2009 strain isolated from Yunnan, China,2009. Eight overlapping clones covering the whole viral genome (excluding the poly-A tail) were obtained by RT-PCR and sequenced, and their nucleotide and amino acid sequences were compared with other known CVB1 strains. The genome of the CVB1 MSH/KM9/2009 strain had 7384 nucleotides in length, and contained a 741nt non-translated region (NTR) at the 5' end and a 94nt NTR at the 3' end. The entire open reading frame contained 6 549 nt, encoding a 2 183-aa polyprotein. In the coding region, there was no nucleotide deletion or insertion, but some changes of amino acid were unique. The complete genome sequence alignments showed that the CVB1 isolate MSH/KM9/2009 strain shared the highest nucleotide (80.9%, 81.6%, 80.5% and 80%) and amino acid (95.6%, 95.8%, 96.2% and 95.6%) identities to the CVB1 M16560, pmMC, Tucson B1 and CVB1Nm strain, respectively. Phylogenetic tree analysis showed that the MSH/KM9/2009, CVB1 M16560, pmMC, Tucson B1 and CVB1Nm strain clustered into same group. The newly isolated CVB1 strain MSH/KM9/2009 from Yunnan Province belonged to genotype CVB1.
Base Sequence ; Child, Preschool ; China ; Coxsackievirus Infections ; virology ; Enterovirus ; classification ; genetics ; isolation & purification ; Female ; Genome, Viral ; Genotype ; Humans ; Infant ; Male ; Open Reading Frames ; Phylogeny ; Viral Proteins ; genetics

To analyze the genomic sequence characteristics of a human Echovirus 9(ECHO-9) strain isolated from a child with Hand-foot-mouth disease (HFMD) in Kunming, Yunnan Province, in 2010. The complete genome sequence of a human echovirus 9 strain, MSH-KM812-2010 was determined. As other human enterovirus, its genome was 7,424 nucleotides (nts) in length and encoded for 2,203 amino acids (aas). In comparison to other human enteroviruses, MSH-KM812-2010 strain had the highest homology with other strains of human echovirus 9 in structural genomic regions and more homologous to other serotypes of B specie than to human echovirus 9 in non-structural genomic regions. Phylogenetic analysis based on complete VP1 gene revealed that the sequences of human echovirus 9 segregated into three distinct clades A, B and C with more than 15. 0% diversity between clades. All Chinese isolates belonged to the same clade. RDP3 and Blast revealed evident recombination in non-structural genomic regions. This report is the first to, describe the complete genome of the human echovirus 9 in China and provide an overview of the diversity of genetic characteristics of a circulating human echovirus 9.
Base Sequence ; China ; Echovirus 9 ; classification ; genetics ; isolation & purification ; Female ; Genome, Viral ; Humans ; Infant ; Molecular Sequence Data ; Phylogeny ; Viral Proteins ; genetics

OBJECTIVETo study the clinicopathologic features, immunophenotype and gene rearrangement of primary cutaneous diffuse large B-cell lymphoma, leg type (PCLBCL).

METHODSSeven cases of PCLBCL were enrolled into the study. Clinicopathologic analysis, immunohistochemical staining and gene rearrangement for IgH and Igκ were undertaken in the study.

RESULTSAll the seven cases were male, and the median age was 72 years. Patients usually presented with multiple purple tumors, nodules, papules and infiltrative plaques. Two patients had a history of leg injury before onset, and one had mosquito bites. Histologically, the tumor involved the dermis and subcutis with dense and diffuse infiltrative pattern composing of centroblasts and/or immunoblasts. Immunohistochemical staining showed that seven cases (7/7) expressed CD20, six (6/6) expressed bcl-2, four (4/4) expressed MUM-1, four (4/5) expressed CD79a, four (4/5) expressed PAX-5 and four (4/6) expressed bcl-6, respectively. All cases did not express CD3ε, CD45RO, CD10 and CD30. IgH gene rearranged bands were detected in three (3/6) cases and Igκ was detected in one (1/5) case. Six of the seven cases died and the remaining patient, who was 44-year-old, was alive after 22 months of follow-up.

CONCLUSIONSPCLBCL is rare, predominantly affects elderly male patients. PCLBCL has poor prognosis and high mortality, but younger patients seem to have better prognosis. Some cases had a history of trauma or mosquito bites. The relationship between the history and the onset of PCLBCL needs further evaluation.

Aged ; Aged, 80 and over ; Animals ; Antigens, CD ; analysis ; Culicidae ; Gene Rearrangement ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin kappa-Chains ; genetics ; Immunophenotyping ; Insect Bites and Stings ; complications ; Leg ; Leg Injuries ; complications ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; Proto-Oncogene Proteins c-bcl-6 ; metabolism ; Skin Neoplasms ; genetics ; pathology

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