To explore the effects of Shenmai Injection (SMI), a compound traditional Chinese herbal medicine, on pharmacokinetics and serum concentration of digoxin when applied together with digoxin.

BACKGROUNDDiffusion-weighted imaging has been widely used to differentiate the character of lymphadenopathy. But there are significant differences between prior studies. The aim of the study was to compare the benefit of apparent diffusion coefficient (ADC) and rADC in the differentiation of metastatic and benign lymph nodes in a rabbit model.

METHODSTwo observers independently measured ADCs in quadriceps of every rabbit on diffusion-weighted images with different sizes of regions of interest (ROI). The appropriate ROI for rADC was determined using the interobserver coefficient. rADC was calculated by ADC(lesion)/ADC(reference site). Receiver operating characteristics (ROC) analysis was performed to compare the diagnostic value of the ADC values and rADC values in differentiating metastatic from benign lymph nodes.

RESULTSWhen the ROIs included five pixels, the ICC was 0.816, indicating a good interobserver agreement. The differences of ADC and rADC values between metastatic and benign lymph nodes were both statistically significant. The area under the ROC curve was greater for the rADC than for the ADC. With the rADC criteria of 0.640, the sensitivity and specificity for differentiating metastatic from benign lymph nodes were 93% and 90%, respectively.

CONCLUSIONAs a reference site, muscles reveal a good repeatability using a small ROI and the rADC may improve diagnostic accuracy for detecting metastatic nodes in animal models.

Animals ; Diffusion Magnetic Resonance Imaging ; methods ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; Rabbits ; Sensitivity and Specificity

OBJECTIVEThis study aimed at exploring the feasibility of using dried blood spots (DBS) to detect HIV drug resistance genotyping in China by comparing the results of drug resistance from DBS, plasma and whole blood samples.

METHODSBlood samples were collected from 39 AIDS patients from Anhui (10), Yunnan (13), Hunan (6) and Xinjiang (10) provinces and autonomous regions. The HIV strains that infected these patients covered all the major HIV-1 subtypes prevailing in China (B, CRF01_AE, CRF07_BC). HIV drug resistance genotyping assay was performed on DBS as well as on the whole blood and plasma samples from the same patients simultaneously by using an in-house nest RT-PCR method. Drug resistance levels were determined based on Stanford University HIV drug resistance database, and the results from these three types of samples were compared.

RESULTSThe percentages of successful amplification of protease and reverse transcriptase regions in the pol gene were 95% (37/39) from DBS, 92% (36/39) from whole blood and 100% (39/39) from plasma samples. The sequences from the three types of samples showed more than 99% identity.86% (31/36) of the DBS samples had the same set of drug resistance mutations as those which were detected from plasma samples. The differences probably resulted from mixed bases.

CONCLUSIONSThere was no major difference in detecting HIV drug resistance genotyping among DBS, plasma and whole blood samples. Therefore, DBS is useful for detection of HIV drug resistance genotyping and is particularly valuable in developing countries like China, especially in remote rural regions.

Dried Blood Spot Testing ; Drug Resistance, Viral ; genetics ; Feasibility Studies ; Genotype ; HIV Infections ; blood ; genetics ; virology ; HIV Seropositivity ; blood ; genetics ; virology ; HIV-1 ; drug effects ; genetics ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Load

OBJECTIVETo summarize the clinical characteristics of secondary coagulation disorders caused by exposure to poison (raticide) in children and to investigate the diagnosis and corresponding treatment.

METHODThe process of diagnosis, clinical characteristics, response to treatment and the prognosis were analyzed.

RESULTSThe main clinical manifestation was mucosal bleeding (66.6%), including epistaxis, gingival bleeding, hematomas and so on. All these children were previously well and had no history of bleeding. Activated partial thromboplastin time (APTT) and prothrombin time (PT) were prolonged, factor II was undetectable and the levels of factors VII, IX, and X were lower. The fibrinogen was normal. A raticide was detected in blood and urine of 13 children although 12 of the patients had no definite history of raticide ingestion. Prothrombin complex, fresh frozen plasma and vitamin K(1) were effective in these cases. However, 2 - 3 weeks later, 6 patients presented with recurrent bleeding.

CONCLUSIONFor children with secondary coagulation disorders of unknown cause, intoxication of raticide should be considered. The administration of blood coagulation factors and vitamin K(1) are effective in early treatment, and the treatment period should be more than 2 months. The PT and APTT should be followed up. Vitamin K(1) should be stopped when PT and APTT are normal.

Blood Coagulation Disorders ; chemically induced ; diagnosis ; therapy ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Rodenticides ; poisoning ; Vitamin K 1 ; administration & dosage ; therapeutic use

PURPOSE: Cutaneous lymphocyte-associated antigen (CLA)-expressing CD8+T cells have been known to play an important role in the pathogenesis of atopic dermatitis (AD). However, the mechanisms underlying the loss of self-tolerance remain unclear. Regulatory T cells (Tregs) play a key role in the development of homeostasis in the immune system. We, therefore, hypothesized that a reduced ability of Tregs to inhibit autologous CD8+CLA+T cells might be underlying mechanism in AD. MATERIALS AND METHODS: CD8+CLA+T cells and Tregs were obtained from the peripheral blood of AD patients and control volunteers. The frequencies of CD8+CLA+T cells were evaluated. The proliferative responses of CD8+CLA+T cells were assessed by flow cytometry, and the levels of transforming growth factor-beta1 (TGF-beta1) and interleukin-10 (IL-10) in culture supernatants were detected by enzyme-linked immunosorbent assay. RESULTS: Our results revealed higher frequency and increased expression of perforin and granzyme-B in peripheral CD8+CLA+T cells in AD, and lower inhibitory ability of Tregs on proliferation of CD8+CLA+T cells in AD. Meanwhile, the levels of TGF-beta1 produced by Tregs were significantly lower in AD, and anti-TGF-beta1 abolished such suppression. CONCLUSION: The attenuated inhibitory ability of Tregs on hyper-activated autologous CD8+CLA+T cells, mediated by TGF-beta1, plays an important role in the pathogenesis of AD.
Adult ; Aged ; CD8-Positive T-Lymphocytes/drug effects/*immunology ; Case-Control Studies ; Cell Proliferation ; Cell Separation ; Dermatitis, Atopic/*immunology/pathology ; Female ; Granzymes/metabolism ; Humans ; Interleukin-10/metabolism ; Lymphocyte Count ; Male ; Perforin/metabolism ; Skin/*immunology/pathology ; T-Lymphocytes, Cytotoxic/drug effects/immunology ; T-Lymphocytes, Regulatory/drug effects/*immunology ; Transforming Growth Factor beta1/pharmacology

Objective To explore the expression of FAT1 in esophageal squamous cell carcinoma ( ESCC) tissues, and its effect on cell proliferation. Methods The expression levels of FAT1 protein in human ESCC tissues and matched adjacent normal tissues were determined by immunohistochemistry ( IHC) . Lentivirus based knockdown of FAT1 was carried out in YSE2 and Colo680N cell lines and 3?( 4,5?dimethyl?2?thiazolyl)?2,5?diphenyl?2H tetrazolium bromide ( MTT) assays was performed to examine the effect of FAT1 on the proliferation of these ESCC cells. Colony formation assay was used to detect the colony formation ability. Flow cytometry was performed to analyze the cell cycle and apoptosis. The expression levels of cell cycle markers in FAT1 knock out ESCC cell lines were detected by real?time quantitative reverse transcription polymerase chain reaction( qRT?PCR) and Western blot. Results The relative expression of FAT1 in ESCC tissues was 66. 97 ± 21. 53, significantly lower than 78. 13 ± 16. 76 of adjacent normal tissues ( P<0.05) . Knockdown of FAT1 promoted cell proliferation and colony formation. In YSE2 cell, the division time in negative control (NC) group was (1570±51) min, significantly longer than (1356±31) min in shFAT1 group. In Colo680N cell, division time in NC group was (1532±53) min, significantly longer than (1290±30) min in shFAT1 group (P<0.05). Knockdown of FAT1 promoted G1?to S?phase transition and resulted in the upregulation of CDK4/CDK6/CCND1. Conclusion FAT1 inhibits the proliferation and G1?to S?phase transition of ESCC cells through regulating the protein expression of CDK4/CDK6/CCND1 complex.

Objective To explore the expression of FAT1 in esophageal squamous cell carcinoma ( ESCC) tissues, and its effect on cell proliferation. Methods The expression levels of FAT1 protein in human ESCC tissues and matched adjacent normal tissues were determined by immunohistochemistry ( IHC) . Lentivirus based knockdown of FAT1 was carried out in YSE2 and Colo680N cell lines and 3?( 4,5?dimethyl?2?thiazolyl)?2,5?diphenyl?2H tetrazolium bromide ( MTT) assays was performed to examine the effect of FAT1 on the proliferation of these ESCC cells. Colony formation assay was used to detect the colony formation ability. Flow cytometry was performed to analyze the cell cycle and apoptosis. The expression levels of cell cycle markers in FAT1 knock out ESCC cell lines were detected by real?time quantitative reverse transcription polymerase chain reaction( qRT?PCR) and Western blot. Results The relative expression of FAT1 in ESCC tissues was 66. 97 ± 21. 53, significantly lower than 78. 13 ± 16. 76 of adjacent normal tissues ( P<0.05) . Knockdown of FAT1 promoted cell proliferation and colony formation. In YSE2 cell, the division time in negative control (NC) group was (1570±51) min, significantly longer than (1356±31) min in shFAT1 group. In Colo680N cell, division time in NC group was (1532±53) min, significantly longer than (1290±30) min in shFAT1 group (P<0.05). Knockdown of FAT1 promoted G1?to S?phase transition and resulted in the upregulation of CDK4/CDK6/CCND1. Conclusion FAT1 inhibits the proliferation and G1?to S?phase transition of ESCC cells through regulating the protein expression of CDK4/CDK6/CCND1 complex.

Objective To evaluate the amplification rate and the lowestlower detection limit of an in-house HIV-1 Drug resistant (HIVDR) genotyping test.Methods A total of 30 plasma samples were selected,which covered allmajor HIV-1 subtypes predominating prevailing in china (B',CRF07_BC,CRF01 _AE ).The viral loads of the 30 selected samples were detected in triplicate by Easy Q method and the average values were taken as the viral loads of the samples.Each sample was diluted to the concentration of ＞ 1000copies/ml,401-1000 copies/ml,101-400 copies/ml,50-100 copies/ml and ＜ 50 copies/ml with HIV-negative plasma. After extraction of nucleic acids, RT-PCR and nested PCR amplification were performed,the efficiency of amplification of each subtype and the minimum detection limit were determined statistically based on the PCR results.Results The viral loads of the selected samples ranged from 2.03 × 102 - 5.92 × 104 copies/ml.The sample of 50 - 1000 copies/ml have a high amplification rate (86％).Conclusion The In-house method for HIV-1 drug resistance genotyping has a high sensitivity with a high successful amplification rate,especially in the samples with low viral load.This method can be used to the detectionof drug-resistant virus and to provide scientific data to treatment options for patients.

To explore the possible mechanism of liver injury, the effects of Ploygoni Multiflori Caulis and its extractive on the function of bilirubin-associated transporters were investigated in normal (N) and idiosyncratic (LPS) rats (M). The normal and LPS rats were respectively administrated powder of Ploygoni Multiflori Caulis, its extractive and same volume of 0.5% CMC-Na solution for 7 d. BSP, a substrate of the transporters of Oatp1a1 and Oatp1b2 was selected, and its pharmacokinetic parameters of intravenous injection were determined to examined the activity these transporters. Meanwhile the mRNA expressions of transporters were detected. Compared with N-blank control group, besides M-powder group, the Cmax has no significantly different from other groups, t1/2, AUC0-t and AUC0-∞ were significantly increased, and CL were significantly decreased. However, compared with N- blank control group, AST and ALT decreased significantly. The expression of Oatp1a1, Oatp1b2 and MRP2 mRNA was significantly decreased (P<0.05), but there was no act synergistically when Ploygoni Multiflori Caulis and extractive were combined with LPS. The function of Oatp1a1, Oatp1b2 and MRP2 in rats were significantly inhibited by Ploygoni Multiflori Caulis and extractive, which may be an important cause of hepatotoxicity.

MicroRNAs (miRNAs) are known to regulate post-transcriptional gene expression. They are involved in carcinogenesis and tumor progression. The aim of this study was to explore the microRNA-mRNA regulatory network in esophageal squamous cell carcinoma (ESCC) using comprehensive computational approaches. In this study we have selected a total of 11 miRNAs from one previously reported study in ESCC. The mRNA targets of these miRNAs were predicted using various algorithms. The expression profiles of these mRNA targets were identified on DNA microarray experiment dataset across ESCC tissue samples. Based on the miRNA-mRNA regulatory relationships, the network was inferred. A total of 23 miRNA-mRNA regulatory interactions, with 11 miRNAs and 13 mRNA targets, were inferred in ESCC. The miRNA-mRNA regulatory network with increased confidence provides insights into the progression of ESCC and may serve as a biomarker for prognosis or the aggressiveness of ESCC. However, the results should be examined with further experimental validation.
Carcinoma, Squamous Cell ; genetics ; Case-Control Studies ; Esophageal Neoplasms ; genetics ; Gene Regulatory Networks ; Humans ; MicroRNAs ; genetics ; RNA, Messenger ; genetics