Objective To evaluate the diagnostic value of fluorescent quantitative polymerase chain reaction(PCR) for Mycoplasma pneumoniae (MP) in children with MP pneumonia(MPP).Methods From Jun.2008 to Jan.2009,153 cases hospitalized with pneumonia were enrolled,and 30 cases without respiratory infection were enrolled as control group.Their respiratory secretion (including nasopharyngeal secretion,sputum,bronchialalveolar lavage fluid or pharyngeal swab) samples were collected for fluorescent quantitative PCR for MP.And their single or paired serums were collected for specific MP antibody detection.Results There were 123 cases confirmed with MPP by serology,among whom 114 cases were MP PCR positive.The quantitation of MP DNA was among 1.20?106-3.66?1010 gene copys/L. There were 30 cases with pneumonia negative with MP by the paired serum serology,among whom 2 cases were MP PCR positive,and the quantitation of MP DNA was (1.08-3.02)?107gene copys/L.All cases of control group were MP PCR negative.During the first and second weeks of the MPP onset,the sensitivity of MP-IgM from the first single blood samples were 66.7% and 83.9%,respectively.While the sensitivity and specificity of MP PCR were 92.7% and 93.3%,respectively.From the third week of the disease onset,the sensitivity of MP-IgM from the first single blood samples increased to 90.9%-100%.The clinical manifestations of MPP were nonspecific.Conclusions PCR is superior to serology for early diagnosis on MP infection.Combination of the 2 methods may be helpful to early and accurate diagnosis on MP infection.

Objective To investigate the effects of enteral nutrition with galactooligosaccharides (GOS) on serum inflammatory cytokines in rats with severe acute pancreatitis (SAP). Methods SD rats were randomly divided into sham operation control group, SAP with enteral nutrition (EN) group and SAP with EN supplemented with GOS (GOS-EN) group, and each group was divided into 4 d and 7 d subgroups according to the time that animals were sacrificed (n=8 in each subgroup). Rat SAP models were established by injection of 38 g/L sodium taurocholate beneath the pancreatic capsule. The serum amylase, inflammatory cytokines TNF-α, IL-2 and IL-10 were detected. Results At each time point, the levels of serum amylase in all SAP groups were significantly higher than those in sham operation control group (P < 0.01), and the levels in GOS-EN group were significantly lower than those in EN group (P < 0.01). The levels of serum TNF-α and IL-10 in all SAP groups were significantly higher than those in sham operation control group (P < 0.01), while the levels of IL-2 in all SAP groups were significantly lower than those in sham operation control group. The levels of TNF-α in GOS-EN group were significantly lower than those in EN group (P <0.05), while the levels of IL-2 and IL-10 and the ratio of IL-10/TNF-α were significantly higher than those in GOS-EN group (P < 0.05). Conclusion Early EN supplemented with GOS could modulate the balance of pro- and anti-inflammatory response.

Objective To analyze the clinical features,laboratory results,treatment and prognosis of children with idiopathic pulmonary hemosiderosis (IPH).Methods The documents of 25 children with IPH,who were hospitalized in Children's Hospital Affiliated to Capital Institute of Pediatrics from Apr.1992 to Nov.2008 were reviewed.Then some of the patients were followed up.Results The median age of onset was 4 years old (6.5 months to 8 years old).Those patients aged from 3 to 6 years old ranked the first as 48％ (12 cases).The median age at diagnosis was 5.17 years old.The course of desease was between 10 days and 6 years,the median course was 1 year.Cough,pallor,fever and hemoptysis were the common clinical features of IPH patients.The chest radiological patterns for IPH were diverse.The common features of high resolution CT scan in 15 patients included declined transparency and ground glass shadows in 11 cases,cloudy patchy infiltrate in 9 cases,reticular changes in 6 cases,and nodular changes in 2 children.Twentythree cases were once misdiagnosed and 60％ of them were delayed in diagnosis as IPH for more than 1 year.Glucocorticoid therapy was effective in improving symptoms.Some patients suffered from rheumatoid arthritis later in their life.Conclusions The manifestations of IPH in children are nonspecific,therefore it is easily to be misdiagnosed.Combined chest radiographic presentations with repeatedly looking for hemosiderin-laden macrophages in sputum,gastric aspirate or bronchoalveolar lavage fluid are helpful in diagnosis.Glucocorticoid therapy can control the symptoms.Some relationships may exist between IPH and rheumatoid arthritis.

Objective·To extract serum IgA1 from patients with IgA nephropathy (IgAN) (end stage renal disease vs long-term stable renal function) to explore the effect on proliferation rate of human mesangial cells (HMCs) and the level of transforming growth factor-β1 (TGF-β1). Methods?·?Nine patients with primary IgAN were divided into rapidly progressive group (n=6) and long-term stable group (n=3). Jacalin affinity chromatography and sephacryl S-200HR molecular sieves were used to distract serum IgA1. HMCs were cultured and co-cultivated with different IgA1 concentration (10, 50, 250 and 1?000?μg/mL)at point of 12?h and 24?h respectively. The proliferation rate was measured by cell counting kit-8 (CCK8). The expression of TGF-β1 mRNA was measured via quantitative real-time PCR (qRT-PCR). Enzyme-linked immunosorbent assay (ELISA) was used to detect TGF-β1 protein. Results?·?Serum IgA1 from IgAN patients with different renal functions (end stage renal disease vs long-term stable renal function) activated proliferation of HMC significantly, presenting with time-dependence and concentration-dependence. The highest value showed at 250?μg/mL and 1?000?μg/mL respectively. Serum IgA1 in two groups of patients statistically increased the expression of TGF-β1 mRNA and protein. In group with end stage renal disease, the summit stood at 10?μg/mL and started to decrease by degrees afterwards; while in group with long-term stable renal function, the level of TGF-β1 increased in a parallel manner with the serum IgA1. Conclusion?·?Serum IgA1 from IgAN patients with different renal functions (end stage renal disease vs long-term stable renal function) can both promote the proliferation of HMC. There is no dramatical difference observed with in10-50?μg/mL, but the IgA1 from group with end stage renal disease reveals a stronger effect on TGF-β1, in accordance with the pathological type of the patients (IgA sclerosis), suggesting the prognostic value of serum IgA.

OBJECTIVETo investigate the quantity and function of circulating dendritic cells (DC) in patients with chronic idiopathic thrombocytopenic purpura (ITP).

METHODSHigh dose dexamethasone (HD-DXM) at a dose of 40 mg orally per day for four consecutive days was the initial treatment for chronic ITP patients. Flow cytometry was used to analyze the number of myeloid DC (mDC), plasma cytoid DC (pDC) and CD4+FOXP3+ T cells in patients before and after the treatment, meanwhile the co-stimulatory molecules on circulating DCs were assayed as well. Monocyte-derived DCs and CD4+ T cells were co-cultured with autologous or allogeneic normal fresh platelets and after 6 days of incubation H-TdR was used to assay the proliferation of CD4+ T cells.

RESULTSThe absolute numbers of circulating mDC and pDC were not significantly different between pre-treatment patients and healthy controls (P > 0.05 and P >0.05). However, percentage of CD4+ FOXP3+ T cells was decreased (P < 0.01), and their percentage was inversely correlated with the number of pDC and mDC (r = -0.396, P =0.045 and r = -0.410, P =0.037). The initial response rate to HD-DXM was 92.3%. After 4-days treatment, CD4 FOXP3+ Treg cells increased (P <0.01) while pDCs decreased (P <0.01). Although mDCs increased after HD-DXM (P <0.05), their CD11c expression level was decreased (P < 0.01), the mean fluorescence intensity (MFI) decreased from 340 +/- 30 before treatment to 199 +/- 21 after treatment. The inverse correlation between pDCs and CD4+ FOXP3+ Treg cells remained (r= -0.524, P =0.006) while that between mDCs and Treg cells disappeared (r = - 0.360, P =0.071). The MFI of CD86 on DCs was higher in ITP patients than in healthy controls (P <0.05), while the proportions of CD86, CD40, CD80 and the MFI of CD40, CD80 in ITP patients were normal (P > 0.05). DCs from chronic ITP patients co-cultured with autologous or allogeneic platelets were highly efficient in stimulating autologous CD4+ T cells proliferaton as compared to those derived from healthy donors (P < 0.05 and P <0.05).

CONCLUSIONDCs may play a role in the pathogenesis of chronic ITP in relation with CD4+CD25+ Treg cells.

Adult ; CD4-Positive T-Lymphocytes ; immunology ; Dendritic Cells ; immunology ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; blood ; immunology

The ethyl acetate extract of the flower of Albizia julibrissin was isolated and purified by silica gel, Sephadex LH-20 and MCI GEL CHP-20P column chromatography to yield 29 compounds. Their structures were elucidated as 8-hydroxy-2, 6-dimethyl-2E, 6Z-octadienoic acid (1), 8-O-formyl-2, 6-dimethyl-2E, 6Z-octadienoic acid (la), 8-hydroxy-2, 6-dimethyl-2E, 6E-octadienoic acid (2), 8-O-formyl-2, 6-dimethyl-2E, 6E-octadienoic acid (2a), (2E, 6S)-2, 6-dimethyl-6-O-beta-D-xylpyranosyloxy-2, 7-menthia-folic acid (3), clovan-2beta, 9alpha-diol (4), 2beta-O-formyl-clovan-9alpha-ol (4a), 2beta, 9alpha-O-diformyl-clovan (4b), vomifoliol (5), (6S, 9R)-roseoside (6), vanillin (7), 4-O-ethylgallic acid (8), 3-ethoxy4-hydroxy-benzoic acid (9), p-hydroxybenzaldehyde (10), gallic acid (11), protocatechoic acid (12), stearic acid (13), palmitic acid (14), 2, 3-dihydroxypropyl hexadecanoate (15), linoleic acid (16), scopoletin (17), indole-3-carboxaldehyde (18), 2-furoic acid (19), 5-(hydroxymethyl)-2-furaldehyde (20), (22E, 24R)-5alpha, 8alpha-epidioxy-ergosta-6, 22-dien-3beta-ol (21), (22E, 24R)-5alpha, 8alpha-epidioxy-ergosta-6, 9, 22-trien-3beta-ol (22), (+)-lariciresinol 9'-stearate (23), formononetin (24) and uridine (25). Compounds 1a, 2a, 4a and 4b were new artifacts from the separation process, and others were obtained from A. julibrissin for the first time.
Albizzia ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flowers ; chemistry ; cytology ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

ABSTRACT:OBJECTIVE The aim of this study was to identify Alisma orientale (Sam.)Juzep.and its adulterants using the ITS2 barcode.METHODS The total DNAs were extracted from twenty-eight samples of A.orientale as well as adulterants. The ITS2 sequences of these samples were amplified and sequenced.The acquired sequences were submitted to the GenBank and the other ITS2 sequences of 17 samples from ten species were downloaded from the GenBank.Total 45 ITS2 sequences were aligned and the genetic distances among them were analyzed with MEGA 5.1.The sequences analyses were performed u-sing the BLAST1 and the nearest distance methods,and were presented intuitively by constructing Neighbor-joining(NJ)tree. RESULTS The lengths of all ITS2 sequences of A.orientale and A.plantago-aquatica were 311 bp.Only one mutation exis-ted among them.There was significant divergence between the interspecific and intraspecific genetic distances of the ITS2 se-quences.The NJ tree showed that A.orientale is obviously different from its adulterants,which showed high monophyly. CONCLUSION As a DNA barcode,ITS2 sequences can stably and accurately distinguish A.orientale from its adulterants and also provide a new clue to identify the medical germplasm resources and ensure the clinical safety in utilization of Chinese mate-ria medica.

To assess the quality of randomized controlled trials (RCTs) published in Chinese Acupuncture and Moxibustion. Manually retrieve RCT papers published in Chinese Acupuncture and Moxibustion from 2000 to 2006 and use "the extraction form of acupuncture and moxibustion randomized clinical trial report data" to evaluate the quality of RCTs. Six hundred and eighty-six RCTs were enrolled. The methodological quality was lower than the international standard and only one RCT paper described the trial technological process. And there were a less reports about influencing RCT quality, such as acupuncturist's qualification and education background (3.06%), adverse reaction (4.23%), blind method (5.98%) and follow-up (14.43%). RCTs on acupuncture and moxibustion published in Chinese Acupuncture and Moxibustion need to be improved in trial design, conduction and report.
Acupuncture Therapy ; Humans ; Moxibustion ; Periodicals as Topic ; standards ; statistics & numerical data ; Randomized Controlled Trials as Topic ; standards ; statistics & numerical data ; Time Factors

OBJECTIVETo study the effect of aqueous extract of several kinds of herbs on human platelet aggregation and expression of P-selectin in vitro.

METHODSBlood was collected from volunteers. Effects of the prepared water extracts of herbs on platelet aggregation were monitored on a Packs-4 aggregometer. The fluorescence intensity of water extracts of Caulis Spatholobi, Flos Carthami and Rhizoma Curcumae on the expression of P-selectin in human platelets of healthy persons was measured with flow cytometry.

RESULTSOut of several herbs investigated, Flos Carthami and Rhizoma Curcumae potently inhibited platelet aggregation after incubation with platelet-rich plasma (PRP) for 15 min. Caulis Spatholobi Flos Carthami and Rhizoma Curcumae inhibited adenosine-5'-diphosphate (ADP) or platelet activating factor (PAF)-induced platelet aggregation in PRP in a dose-dependent manner. In contrast to Flos Carthami and Rhizoma Curcumae, Caulis Spatholobi could not inhibit thrombin-induced platelet aggregation. Despite its inability to inhibit thrombin-induced platelet aggregation in PRP, Caulis Spatholobi had a greater anti-aggregating activity in PRP induced by ADP or PAF. Caulis Spatholobi and Flos Carthami showed significant inhibitory effects on the expression of P-selectin.

CONCLUSIONSCaulis Spatholobi, Flos Carthami and Rhizoma Curcumae have potent anti-platelet properties, and their inhibitory actions are mediated via different mechanisms. Caulis Spatholobi inhibited ADP-induced platelet aggregation but not by thrombin, indicating that its mechanism of action might be independent of the thromboxane pathway. The effect of Caulis Spatholobi and Flos Carthami were associated with suppressing the expression of P-selectin.

Adult ; Blood Platelets ; drug effects ; metabolism ; Curcuma ; chemistry ; Fabaceae ; chemistry ; Humans ; P-Selectin ; metabolism ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Function Tests ; Water ; chemistry ; Young Adult

OBJECTIVETo investigate the role of CD4+ CD25+ regulatory T (Treg) cells in patients with ITP.

METHODSFlow cytometry was used to examine the number of CD4+ CD25+, CD4+ CD25 high, CD4+ Foxp3+ and CD4+ CD25+ Foxp3+ T cells. The level of Foxp3 mRNA expression was analyzed by realtime quantitative reverse transcriptase polymerase chain reaction (RT-PCR) . CD4+ CD25 high T cells was cocultured with CD4+ CD25 - T cells from patients or controls for assessing the regulatory properties of CD4+ CD25 Treg cells.

RESULTSThe proportion of CD4+ CD25+ T cells in the peripheral blood of patients with ITP [(15.64 +/- 5.82) %] was significantly higher than that in normal control group [(9.30 +/- 3.95)%] (P <0.01). There was no significant difference in the percentages of CD4+ CD25 high T cells between ITP patients and controls [(1.53 +/- 0.66)% versus (1.36 +/- 0.55)% (P = 0.317)]. But the number of CD4 Foxp 3+ T cells and CD4 + CD25+ Foxp3+ T cells in patients were significantly lower than that in control group (P <0.01). The expression of Foxp3 mRNA reduced (P < 0.01) and the suppressive activity of CD4+ CD25 high T cells is lower than that of healthy controls (P <0.01).

CONCLUSIONIn patients with ITP, both the number and immuno-regulative function of CD4+ CD25+ Treg cells are reduced.

Adolescent ; Adult ; Aged ; Female ; Forkhead Transcription Factors ; metabolism ; Humans ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; metabolism ; RNA, Messenger ; metabolism ; T-Lymphocytes, Regulatory ; immunology