Objective To investigate the effects of baicalin on the apoptosis and cell migration of intestinal epithelial cells 6 (IEC-6) induced by lipopolysaccharide (LPS) . Methods LPS was adopted to establish LPS-induced cells injury model. Methyl thiazolyl tetrazolium ( MTT) assay and Elisa kits were used to evaluate the effect of LPS on the tumor necrosis factor alpha ( TNF-α) and interleukin 6 ( IL-6) concentrations of IEC-6. Hoechst 33258 staining and flow cytometry were used to evaluate the effect of baicalin ( in the dosage of 2.5, 5.0, 10.0 μg/mL ) on the cell apoptosis. Cell migration was observed and calculated on a wounding model of IEC-6 cells induced by a pipette tip. Results Compared with the control group, the survival rate of IEC-6 in 1.0 μg·mL-1 LPS group was much lowered, TNF-α and IL-6 concentrations were significantly increased, the apoptotic rate of IEC-6 was increased and the cell migration activity was significantly decreased ( P<0.05) . Compared with the model group, IEC-6 apoptosis was relieved to various degrees in baicalin groups, the apoptotic rate of IEC-6 was obviously decreased (P<0.05) and cell migration activity was increased significantly (P<0.01) in 10.0 μg·mL-1 baicalin group. Conclusion Baicalin has the capacity of protecting IEC-6 from LPS-induced injury and increasing cell migration activity.

Objective To evaluate the dynamic changes of cell mobility of renal tubular epithelial cells in the course of epithelial-mesenchymal transition(EMT) and their effect on cell cycle.Methods NRK-52E cells were cultured in vitro and treated with 5 μg/L transforming growth factor(TGF)-β1 to induce EMT.The cell mobility was assessed by using Transwell chamber assay and flow cytometry (FCW) after being treated with TGF-β1 for 4 h,8 h,12 h,24 h and 48 h.The proliferative cell cycle of NRK-52E cells were evaluated by using the FCW.Results 1.EMT was successfully induced by TGF-β1.After being treated by TGF-β1 (5 μg/L),the morphological changes of NRK-52E cells were found with loose cell arrangement and elongated fusiform change in cells body.Meanwhile,after getting treated by TGF-β1,the expressions of E-cadherin protein(epithelial marker) of NRK-52E cells were significantly decreased with time-dependent (P ＜ 0.05),while the expressions of α-smooth musle actin (α-SMA) (mesenchymal cell marker)were significantly increased with time-dependent (P ＜ 0.05).2.The Transwell chamber assay showed that compared with the control group,the cell mobility in the group treated with TGF-β1 was significantly enhanced from 12 h after getting treated with TGF-β1 (P ＜ 0.01).3.The proliferative cell cycle of NRK-52E cells showed no significant difference after being treated with TGF-β1 (P ＞ 0.05).Conclusions The migration ability of the NRK-52E cells are increased incessantly in the course of EMT,which is induced by TGF-β1 without the influence of cell proliferation in vitro.

AIM To establish a UPLC-QTOF/MS method for the analyses of chemical constituents and constituents absorbed into blood from Qili Qiangxin Capsules by UPLC-QTOF/MS.METHODS The rats with heart failure after myocardial infarction were given intragastric administration of 0.5%CMC-Na suspension of Qili Qiangxin Capsules(crude drug dosage was 0.9 g/mL)for 4 weeks,after which blood collection was made.The analysis was performed on a 40℃thermostatic Acquity UPLC BEH C18 Column(100 mm×2.1 mm,1.7 μm),with the mobile phase comprising of 0.1%formic acid-acetonitrile(positive ion manner)or water-acetonitrile(negative ion manner)flowing at 0.3 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive and negative ion scanning.RESULTS Total 151 constituents were identified,containing 38 flavonoids,36 terpenoids,23 alkaloids,11 saponins,11 phenylpropanol,10 phenolic acids,4 organic acids,4 cardiac glycosides and 14 others,along with 35 constituents absorbed into blood.CONCLUSION This accurate and stable method can provide data support for the rational clinical application of Qili Qiangxin Capsules,and lay foundation for the research on related effector substances of other Chinese traditional patent medicines.

BACKGROUNDPenicillium marneffei (P. marneffei) is an emerging pathogenic fungus that can cause invasive mycosis in patients with AIDS. The epidemiological features of P. marneffei infection in AIDS patients in Guangdong province remain unclear so far. This study aimed to investigate the genetic diversity within a population of 163 P. marneffei isolates obtained from AIDS patients and search for the dominant clinical strains in Guangdong province.

METHODSOne hundred and sixty-three P. marneffei isolates obtained from AIDS patients in Guangdong province during January 2004 and December 2009 were studied by randomly amplified polymorphic DNA (RAPD) using two random primers (H2 and H22). The degree of similarity between samples was calculated through similarity coefficients from RAPD fragment data and the dendrogram was assessed using the unweighted pair group method with arithmetic mean (UPGMA).

RESULTSTwo primers showed a high degree of discrimination and good stability. Primer H2 yielded eight different patterns (H2-1 to H2-8) among 163 isolates with the discriminatory power being 0.413. Primer H22 identified seven types (H22-1 to H22-7) among 163 isolates with the discriminatory power being 0.467. Genetic similarity coefficients based on RAPD data among 163 P. marneffei isolates ranged from 0.681 to 0.957, 61.96% of which were no less than 0.83. The discriminatory power of the two primers was 0.524. One hundred and sixty-three P. marneffei isolates were clustered into nine distinct groups (groups I to IX) at the similarity coefficient value of 0.83 and group I was the most common, including 101 strains (61.96%).

CONCLUSIONThe RAPD analyses could provide important information as to the degree of genetic diversity and the relationship among clinical P. marneffei isolates, revealing genetic polymorphism and dominant strains.

Acquired Immunodeficiency Syndrome ; microbiology ; Genetic Variation ; genetics ; Humans ; Penicillium ; classification ; genetics ; Random Amplified Polymorphic DNA Technique ; methods

OBJECTIVETo evaluate the relationship of chimerism status of cell subsets with engraftment, occurrence of acute graft versus host disease (aGVHD), graft rejection and disease relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSChimerism status in peripheral blood (PB) and bone marrow (BM) of 65 patients received allo-HSCT were monitored at regular intervals post-transplant. Fluorescence-activated cell sorter (FACS) was used to sort CD3(+)T lymphocytes in 65 cases, CD3(-)CD56(+)CD16(+)NK cells in 52 cases, CD15(+) granulocytes in 32 cases and CD19(+)B lymphocytes in 20 cases post transplants. The chimerism status of different lineage cells was analyzed by polymerase chain reaction amplification of short tandem repeats (PCR-STR).

RESULTSOn day +7, NK-cells donor chimerism (DC 55.5%) was higher than other cell subsets. T lymphocyte was the latest one to reach complete donor chimerism (CDC) with a median on day +21. Patients whose T lymphocytes donor chimerism was more than 70% on day +7 and more than 95% on day +14 had a high risk for acute aGVHD. In all cases except those with ALL, the decreased DC of T lymphocytes were observed before molecular or hematological relapse occurred.

CONCLUSIONSerial and quantitative T cell chimerism analysis provides a reliable and rapid screening method for the early detection of engraftment, graft rejection, disease relapse and occurrence of aGVHD, therefore, is a prognostic tool to identify patients at high risk of aGVHD and disease relapse following allo-HSCT.

Adolescent ; Adult ; Child ; Chimerism ; Female ; Graft Rejection ; immunology ; Graft vs Host Disease ; immunology ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Middle Aged ; Recurrence ; T-Lymphocytes ; immunology ; Young Adult

This study was aimed to investigate the distribution of abnormal clone in marrow cell lineages and apoptosis cells in myelodysplastic syndrome (MDS) with deletion of chromosome 20q. Monoclonal antibodies recognizing myeloid precursors (CD15), erythroid precursors (GPA), T cells (CD3(+)CD56(-)CD16(-)), B cells (CD19), NK cells (CD3(-)CD56(+)CD16(+)) were used to sort bone marrow cells in a MDS patient with del (20q) by fluorescence activated cell sorting (FACS). Annexin V-FITC and PI were used to sort bone marrow Annexin V(+)PI(-) and Annexin V(-)PI(-) cells by FACS. The sorted positive cells were detected by interphase dual-color fluorescence in situ hybridization (D-FISH) using a LSI D20S108 probe (Spectrum Orange) and a Telvysion TM 20p probe (Spectrum Green). FACS and FISH analysis were also performed on the samples from 4 cases with normal karyotype. The results showed that the proportions of MDS clone in the myeloid and erythroid precursors were 70.50% and 93.33% respectively, in the RAEB-1 patient with del (20q) and were obviously higher than that in control group (5.39% and 6.17%). The proportions of abnormal clone in T, B and NK cells were 3.23%, 4.32% and 5.77% respectively and were less than that in control group (5.76%, 4.85%, 6.36%). The percentage of apoptotic cells in the bone marrow nucleated cells was 16.09%. The proportions of MDS clone in Annexin V(+)PI(-) and Annexin V(-)PI(-) cells were 32.48% and 70.11%, respectively. It is concluded that most myeloid and erythroid precursors are originated from the abnormal clone in MDS with del (20q). A little part of apoptotic cells are derived from the abnormal clone.
Apoptosis ; genetics ; Bone Marrow Cells ; metabolism ; pathology ; Cell Lineage ; genetics ; Chromosome Deletion ; Chromosomes, Human, Pair 20 ; Clone Cells ; metabolism ; pathology ; Humans ; Myelodysplastic Syndromes ; genetics ; pathology

[Objective]To explore the evaluation value of ultrasomics based on contrast-enhanced ultrasound (CEUS)imaging in the therapy response of microRNA-122(miR-122)in hepatocellular carcinoma(HCC).[Method]Mice bearing subcutaneous HCC xenografts were injected intratumorally with microRNA-122 mimics(miR-122 mimics) and negative control mimics(NC mimics)in treatment group(n=6)and control group(n=6),respectively. The injec-tions were performed every 3 days for five times.Before each injection,two-dimension ultrasound(2D-US)imaging was performed.At 24 h after the last injection,2D-US and CEUS images of tumors were acquired,and then mice scarified for tumor miR-122 expression analysis by qRT-PCR.To evaluate the therapy response by RECIST,tumor volumes were mea-sured based on each 2D-US image. To analyze the tumor perfusion by mRECIST,perfusion parameters(maximum of intensity,rise time,time to peak,mean transit time,quality of fit)were analyzed off-line based on dynamic CEUS videos using SonoLiver?software. For ultrasomics,CEUS images at 10,30,60,90 second were used for features extraction, respectively. The corresponding ultrasomics formulas were built to evaluate the therapy response for miR-122.[Result]The tumors treated with miR-122 mimics resulted in a(763±60)folds increase in miR-122 levels compared to the tumors in control group(P<0.05).Effectively therapeutic response evaluated by tumor sizes change was detected after the third injection(P<0.05).For assessment using mRECIST,all the parameters of treatment group did not show significant difference from the ones of control group(P>0.05).Analysis using ultrasomics fail to detect different features of the static images of CEUS at 10 s,and models can be successfully built based on the rest of the three phases of CEUS images.The ultrasomics Scores between control group and treatment group were statistically different(P<0.05).The ultrasomics score at 30s were significantly lower than those at 60 s and 90 s,while there was no statistical difference between scores at 60 s and 90 s.[Conclusion]Ultrasomics analysis based on CEUS imaging is a useful method in evaluating the therapy response of miR-122 in HCC,and showed greater value than dynamic perfusion parameter.

Purpose: This study aimed to establish whether computed tomography (CT)–determined sarcopenia is a useful imaging biomarker for postoperative outcome in elderly colorectal cancer (CRC) patients, and construct sarcopenia-based nomograms to predict individual outcomes after surgery. Materials and Methods: CT imaging data of 298 elderly CRC patients who underwent surgery in 2012-2014 were retrospectively analyzed. Skeletal muscle mass was determined by CT, and sarcopenia was diagnosed based on the optimal cutoff value determined by X-tile program. The correlation between sarcopenia and risk of preoperative nutrition and postoperative complications was evaluated. A Cox proportional hazards model was used to determine survival predictors. Sarcopenia-based nomograms were developed based on multivariate analysis, and calibrated using concordance index and calibration curves. Results: A total 132 patients (44.3%) had sarcopenia based on the optimum cutoff values (29.9 cm2/m2 for women and 49.5 cm2/m2 for men). Sarcopenia was an independent risk factor for preoperative nutrition (p < 0.001; odds ratio [OR], 3.405; 95% confidence interval [CI], 1.948 to 5.954) and postoperative complications (p=0.008; OR, 2.192; 95% CI, 1.231 to 3.903). Sarcopenia was an independent predictor for poor progression-free survival (p < 0.001; hazard ratio [HR], 2.175; 95% CI, 1.489 to 3.179) and overall survival (p < 0.001; HR, 2.524; 95% CI, 1.721 to 3.703). Based on multivariate analysis, we produced four nomograms that had better predictive performance. Conclusion CT-determined sarcopenia is a useful imaging biomarker for predicting preoperative nutritional risk, postoperative complications, and long-term outcomes in elderly CRC patients. The sarcopenia-based nomograms can provide a scientific basis for guiding therapeutic schedule and follow-up strategies.

OBJECTIVETo observe the therapeutic effect of eye-needling combined with medication for treatment of ophthalmoplegia and explore the possible mechanism.

METHODSOne hundred and twenty cases were randomly divided into a treatment group and a control group. According to etiological factors, the control group were treated with medication and the treatment group with the medication plus eye-acupuncture at main point ocular muscles. Changes of the rima oculi, the range of ocular movement and the dialopia angle after treatment were recorded and statistically analyzed in the two groups.

RESULTSThe total effective rate was 93.4% and the cured rate was 54.1% in the treatment group, and 74.6% and 18.6% in the control group, with significant difference between the two groups (P < 0. 01).

CONCLUSIONEye-needling combined with medication has an obvious therapeutic effect which is better than simple medication for ophthalmoplegia.

Acupuncture Therapy ; methods ; Adolescent ; Adult ; Aged ; Child ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Ophthalmoplegia ; therapy

BACKGROUNDAttention deficit hyperactivity disorder (ADHD) is one of the most common mental disorders during childhood, characterized by the core symptoms of hyperactivity, impulsivity and inattention and puts great burden on children themselves, their families and the society. Osmotic release oral system methylphenidate (OROS-MPH) is a once-daily controlled-release formulation developed to overcome some of the limitations associated with immediate-release methylphenidate (IR-MPH). It has been marketed in China since 2005 but still lacks data from large-sample clinical trials on efficacy and safety profiles. The aim of this study was to evaluate the effectiveness and safety of OROS-MPH in children aged 6 to 16 years with ADHD under naturalistic clinical setting.

METHODSThis 6-week, multi-center, prospective, open-label study enrolled 1447 ADHD children to once-daily OROS-MPH (18 mg, 36 mg or 54 mg) treatment. The effectiveness measures were parent-rated Inattention and Overactivity With Aggression (IOWA) Conners I/O and O/D subscales, physician-rated CGI-I and parent-rated global efficacy assessment scale. Blood pressure, pulse rate measurement, adverse events (AEs) and concomitant medications and treatment review were conducted by the investigator and were served as safety measures.

RESULTSA total of 1447 children with ADHD (mean age (9.52 ± 2.36) years) were enrolled in this trial. Totally 96.8% children received an OROS-MPH modal dose of 18 mg, 3.1% with 36 mg and 0.1% with 54 mg at the endpoint of study. The parent IOWA Conners I/O score at the end of week 2 showed statistically significant (P < 0.001) improvement with OROS-MPH (mean: 6.95 ± 2.71) versus the score at baseline (10.45 ± 2.72). The change in the parent IOWA Conners O/D subscale, CGI-I and parent-rated global efficacy assessment scale also supported the superior efficacy for OROS-MPH treatment. Fewer than half of 1447 patients (511(35.3%)) reported AEs, and the majority of the events reported were mild (68.2%). No serious adverse events were reported during the study.

CONCLUSIONThis open-label, naturalistic study provides further evidence of effectiveness and safety of OROS-MPH in school-aged children under routine practice.

Adolescent ; Attention Deficit Disorder with Hyperactivity ; drug therapy ; Child ; Delayed-Action Preparations ; Female ; Humans ; Male ; Methylphenidate ; administration & dosage ; adverse effects ; therapeutic use ; Prospective Studies ; Treatment Outcome