Cytomegalovirus ; genetics ; Cytomegalovirus Infections ; virology ; DNA, Viral ; genetics ; Female ; Genotype ; Hepatitis, Viral, Human ; virology ; Humans ; Infant ; Infant, Newborn ; Male ; Polymorphism, Restriction Fragment Length ; Viral Envelope Proteins ; genetics

AIM　To develop a sensitive and specific LC/MS/MS method for determination of amlodipine in human plasma. METHODS　Amlodipine and internal standard 4′-hydroxypropafenone were extracted from plasma using liquid-liquid extraction, then separated on a Zorbax C8 column. The mobile phase consisted of acetonitrile-water-formic acid (75∶35∶1), at a flow-rate of 0.4 mL*min-1. A Finnigan TSQ tandem mass spectrometer equipped with electrospray ionization source was used as detector and was operated in the positive ion mode. Selected reaction monitoring (SRM) using the precursor → product ion combinations of m/z 409 → 238 and m/z 358 → 116 was used to quantify amlodipine and internal standard, respectively. RESULTS　The linear calibration curves were obtained in the concentration range of 0.4-16.0 ng*mL-1. The limit of quantification was 0.4 ng*mL-1. Each plasma sample was chromatographed within 3.7 min. The method was successfully used in several pharmacokinetic studies for amlodipine. More than 1 500 plasma samples were assayed within two weeks. CONCLUSION　The method is proved to be suitable for clinical investigation of amlodipine pharmacokinetics, which offers advantages of specificity, speed, and greater sensitivity over the previously reported methods.

Objective To evaluate the clinical value of sentinel lymph nodes (SLN) in predicting pelvic lymph node status for early cervical squamous cell carcinoma,and approach the clinical significance of SLN detection for guiding radical abdominal trachelectomy (RAT).Outcomes of follow up and fertility were also observed.Methods A total of 31 patients with stage Ⅰ a2-Ⅰ bl squamous cell carcinoma planned to be given RAT and pelvic lymphadenectomy were enrolled.99mTe-labeled phytate was injected before surgery.Intraoperatively,SLN were identified,excised,and submitted to fast frozen section.Systematic bilateral pelvic lymphadenectomy was performed,and then RAT was performed in patients with negative SLN.All nodes were sent for routine pathological examination and immunostained with anti-cytokeratin antibody to detect micrometastases.Results SLN were detected in all patients (100％,31/31).A total of 109 SLN were identified with a mean number of 3.5 per patient.Of these,SLN of 2 patients were positive on frozen sections and proved to be metastasis by final pathologic examination and quitted the RAT.No missed micrometastasis was found using immunohistochemical staining in SLN and other lymph nodes using histologically node-negative cases.No false negative cases was found and the negative value was 100％ (31/31).The sensitivity,accuracy,and false negative rates were 100％,100％,and 0,respectively.Perioperative complications occured in 5 patients including 2 cases of bladder injury and 3 cases of uterine artery injury.No relapses occurred during follow-up.Five of 19 patients with procreative desire conceived pregnancies (4 spontaneous abortion and 1 premature birth) after surgery.Conclusions The identification of SLN using 99mTc-labeled phytate could predict the pelvic lymph node status in early stage cervical cancer.Under the guidance of SLN detection,RAT is a feasible operative modality with well prognosis and low complications for young patients who desire to preserve reproductive function.

AIM: To isolate targeted the peptides that binding and internalizing into endometrial carcinoma cell line EAC. METHODS: The tumor cells were screened five rounds of whole cell screen through the Ph.D.-12 phage display library. Analysis of the displayed peptides specific binding efficiency to the tumor cells was proceeded. The DNA of phages was extracted, sequenced and translated to the sequences of amino acid and analysis using computer software. RESULTS: After five biopannings, the isolated phages showed high specificity and strong affinity for their cognate cell types relative to different cell lines. Through sequencing, the sequences of displayed peptides were obtained. CONCLUSIONS: Whole cell screening against EAC cells through random phage peptide library can obtain phage peptides with a highly tissue specific binding and internalizing ability. The peptides do provide a basis for tumor's targeted delivery as therapy vector.

Objective To provide an experimental evidence for the clinical applications of glyceryl trinatrate(GTN)and isosorbide dinitrate(ISDN)against Coxsackievirus B(CVB)-related myocarditis.Methods Coxsackievirus B3 was propagated in HeLa cells.Virus yields were determinded by 50% tissue culture infective dosage(TCID50).BALB/c mice were attacked with 5000 TCID50 of CVB3,meanwhile,the mice were administrated with GTN and ISDN.All mice were killed at the 14th day.The myocardial tissues were harvested for histologic evaluation.Results The infection plaques in the myocardial tissues obtained from CVB3-infected BALB/c mice treated with GTN were siginificantly reduced(0.89?0.18 in GTN group and 1.25?0.22 in ISDN group)compared with that of the untreated CVB3-infected mice(P

Objective To detect the expression of stem cell marker Sox2 in gastric cancer (GC). Methods The mRNA and protein expressions of Sox2 in paired primary tumor tissues and their matching, adjacent non-cancerous tissues in a series of 60 cases of human GC were examined by reverse transcription-PCR (RT-PCR) and immunohistochemistry (IHC). χ2 test was used to analyze the correlation of Sox2 expression with clinicopathological parameters of GC tissues including age, gender, tumor size, histological type, TNM stage, differentiation degree, depth of invasion and lymph node metastasis. Results RT-PCR results showed that the positive rate of Sox2 expression was significantly increased in gastric tumor tissues (53.3%, 32/60) compared with that of matching, adjacent non-cancerous tissues (20.0%, 12/60, P<0.01). Semi-quantitative analysis showed that the relative intensity of Sox2 mRNA expression was significantly higher in gastric cancer tissues (0.724±0.209) than that in tissues adjacent to carcinoma (0.256±0.065,P<0.01). The positive expression of Sox2 was significantly higher in gastric tumor tissues (50.0%, 30/60) than that of matching, adjacent non-cancerous tissues (16.7%, 10/60,P<0.01). The positive expression of Sox2 was significantly higher in gastric tumor patients with TNM stage (Ⅲ+Ⅳ) than that of TNM stage (Ⅰ+Ⅱ). The positive expression of Sox2 was significantly higher in gastric tumor patients with low differentiation and undifferentiated tumor cells than that of patients with middle and high differented cells. The positive expression of Sox2 was also significantly higher in gastric tumor patients with the depth of invasion T3-T4 than that of patients with T1-T2. The positive expression of Sox2 was significantly higher in gastric tumor patients with lymph node metastasis than that of patients without lymph node metastasis (P<0.05 or P<0.01). Conclusion The elevated expression of Sox2 is associated with the initiation, invasion, progression, and metastasis of GC. Sox2 may serve as a novel diagnostic and therapeutic marker for human GC.

A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.
Chromatography, Liquid ; Humans ; Tandem Mass Spectrometry ; gamma-Aminobutyric Acid ; blood

OBJECTIVETo observe the therapeutic effect of preconditioning acupuncture (PA) on the treatment of primary dysmenorrhea.

METHODSEighty patients suffered from primary dysmenorrhea were randomly assigned on the ratio of 1:1:2 into three groups, 20 in Group A, 20 in Group B, and 40 in Group C. Group A and B were treated with PA and immediate acupuncture (IA), respectively, while Group C received no acupuncture treatment and be taken as a blank control. The treatment was lasted for three menstrual cycles. The therapeutic effects were observed and compared.

RESULTSComparing the scores of the symptoms in the different groups, the therapeutic effect in different groups showed that the symptom scores in Group A at the second and third cycle of treatment, as well as at the first and third cycle of follow-up, were significantly lower than the scores in Group B, respectively (P<0.05 or P<0.01), and the duration of symptoms was shorter in Group A than in Group B at the third cycle of treatment and the first and third cycle of follow-up (P<0.05). Symptom scores in Groups A and B in the treatment and follow-up periods were all lowered as compared to those before treatment (P<0.01), and the scores were also lower than those in Group C measured at corresponding times (P<0.01).

CONCLUSIONThe therapeutic effect of PA, either the short-term or the long-term effect, was superior to that of IA in treating dysmenorrhea.

Acupuncture Therapy ; methods ; Adolescent ; Adult ; Age of Onset ; Dysmenorrhea ; diagnosis ; epidemiology ; pathology ; therapy ; Female ; Follow-Up Studies ; Humans ; Prognosis ; Research Design ; Severity of Illness Index ; Time Factors ; Treatment Outcome ; Young Adult

OBJECTIVE Toobservetheeffectoftemozolomide(TMZ)incombinationwithtetran-drine(TET)on cell viability,colony formation,migration and cell apoptosis of human glioblastoma U87 cells.METHODS TheviabilityofU87cellstreatedwithTET(8-64μmol·L-1),TMZ(50-400 μmol·L-1 )and TMZ combined with TET (3.2,6.4 μmol·L-1 )was detected by cytotoxicity assays with Cell Counting Kit-8 (CCK-8),the colony formation was detected by Giemsa staining,cell migration ability was detected by Transwell migration assay,cell apoptosis was assayed by flow cytometry using Annexin Ⅴ /PI double staining,and the expression of apoptosis-related proteins expression was detec-tedbyWesternblotting.RESULTS ThedataofCCK-8showedthatTET(r=0.903,P<0.05)orTMZ (r=0.995,P<0.05)could inhibit U87 cell viability alone in a concentration-dependent manner.The cell viability inhibition rate of U87 cells by TMZ co mbined with TET was higher than by TMZ or TET alone. Data showed that the effect of TMZ combined with TET was additive.TMZ 100 μmol·L-1 inhibited U87 cell colony formation and migration ablility compared with normal control.The inhibition rate of U87 cells by TMZ 100 μmol·L-1 combined with TET (3.2 and 6.4 μmol·L-1 )was more significant than by TMZ alone (P<0.05).Compared with TMZ alone,TMZ combined with TET (3.2 and 6.4 μmol·L-1 )signifi-cantly down-regulated the expression of anti-apoptotic protein Bcl-XL,but significantly up-regulated the expression of cleaved caspase 3 protein and cleaved poly(ADP-ribose)polymerase.CONCLUSION TET combined with TMZ can inhibit U87 cell viability,colony formation and migration by activating caspase-dependent apoptotic pathway,resulting in apoptosis.

A chiral LC-MS/MS method for the simultaneous analysis of desvenlafaxine (DVS) enantiomers in human plasma was developed and applied to a pharmacokinetic study on 12 Chinese healthy volunteers. d6-Desvenlafaxine was used as internal standard (IS). Chromatographic separation was performed on the Astec Chirobiotic V chiral column (150 mm x 4.6 mm, 5 μm). The assay was linear over the concentration range of 0.500-150 ng x mL(-1) for both enantiomers (r2 > 0.99). The method was successfully applied to a stereoselective pharmacokinetic study of 100 mg desvenlafaxine sustained release tablets on 12 Chinese healthy volunteers under fasting conditions. The results showed that the pharmacokinetic parameters were similar to both enantiomers in Chinese healthy volunteers. The AUC(0-t), and C(max) of the two enantiomers were about 1.5 times higher than those of blacks and whites reported in the literature.
Administration, Oral ; Area Under Curve ; Asian Continental Ancestry Group ; Chromatography, Liquid ; Cyclohexanols ; blood ; pharmacokinetics ; Delayed-Action Preparations ; Desvenlafaxine Succinate ; Dose-Response Relationship, Drug ; Healthy Volunteers ; Humans ; Male ; Plasma ; chemistry ; Stereoisomerism ; Tablets ; Tandem Mass Spectrometry