Brucellosis ; diagnosis ; drug therapy ; Child ; Child, Preschool ; Female ; Humans ; Male

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of butoconazole in human plasma. Human plasma samples of 0.2 μL were pretreated by a single step protein precipitation procedure and analyzed using a high performance liquid chromatography (HPLC) electrospray tandem mass spectrometer system. The compounds were eluted isocratically on an Inertsil ODS-SP column (100 mm×2.1 mm, 3 μm), ionized using a positive ion atmospheric pressure electrospray ionization source and analyzed using multiple reaction monitoring (MRM) mode. The ion transitions monitored were m/z 412.8→165.1 for butoconazole and m/z 453.4→230.3 for the internal standard. The chromatographic run time was 3.5 min per injection, with retention time of 2.47 min and 2.15 min for butoconazole and repaglinide, respectively. The method was validated to be linear over the range of 20 to 8000 pg/mL (r>0.999) by using a weighted (1/x(2)) quadratic regression. The mean recovery rate was more than 86.7%, and the intra- and inter-day precision of the quality control samples (QCs) was less than 8.3% and the accuracy ranged from 96.0% to 110.2%, which indicated that the quantitative method was reliable and accurate. The method is simple, rapid, and has been applied successfully to a pharmacokinetics study of butoconazole nitrate suppositories in healthy Chinese females.

ObjectiveTo explore the effect of metabolic syndrome (MS) on the occurrence and development of benign prostate hyperplasia (BPH).Methods 101 elderly BPH patients were divided into two groups:BPH (n = 45) and BPH with MS (n= 56)group.The effects of metabolic indexes,including body mass index (BMI),waist,high density lipoprotein cholesterol (HDL-C),fasting blood glucose (FBS) and insuline resistance index (H()MA-IR),on prostate volume(PV),prostate-specific antigen (PSA),international prostate symptom score (IPSS) and lower urinary tract symptoms (LUTS) were surveyed in BPH patients.Results BPH with MS group showed significantly higher values of PV (t = 3.22,P= 0.003)and longer course of LUTS (t= 2.02,P =0.046) than BPH group.The BPH patients with overweight and obesity had significantly higher levels of PV(49.44±26.83 ml and 51.7±22.2 ml,P=0.021 and 0.043) than BPH patients with normal weight (38.10 ± 10.64 ml).Additionally,BPH patients with abdominal obesity had significantly higher levels of PV than BPH patients without abdominal obesity(50.26±26.51 ml vs.38.99± 11.25ml,P=0.005).BPH patients with low HDL-C had significantly higher PV than BPH patients with normal HDL-C[(54.23±28.92)ml vs.(40.40± 14.87) ml,P=0.009].The values of PV,PSA in the BPH patients with elevated FBS were significantly higher than in BPH patients with normal FBS (t=3.17 and 2.4I,P= 0.035 and 0.013).BPH patients with insuline resistance (IR) had higher values of PV and longer courses of LUTS than BPH patients without IR (t= 3.43 and 3.58,P-0.001).The PV was positively correlated with BMI(r= 0.459.P= O.OOO),FINS (r= 0.42,P=O.OOI),HOMA-IR (r= 0.49,P= 0.003) and gatively correlated with HDL-C (r= 0.38,P-0.000)- Multiple linear stepwise regression analysis showed that PV was closely correlated with HOMA-IR.ConclusionsMS has evident effects on the occurrence and development of BPH.

Objective:To prepare rabbit polyclonal antibody against human argonaute 3(AGO3) protein,to identify its properties and investigate the tissue distribution of AGO3 using tissue array.Methods:AGO3 peptide was synthesized using chemical method,and then conjugated to Keyhole limpet hemocyanin(KLH) as immunogen.The AGO3-KLH conjugate were injected into rabbits subcutaneously to produce polyclonal antibodies.The specificity and sensitivity of antibodies were identified by ELISA and Western blot after purification using affinity chromatography.Then the distribution of AGO3 in tissues was examined through immuno-stainning by tissue array.Results:Rabbit polyclonal antibodies against AGO3 were raised after immunization with AGO3-KLH conjugates.The anti-serum titer after the last inoculation was up to 1∶20 000.The preparations of the antibody were confirmed to raised recognize AGO3 peptides specially by ELISA and Western blot.AGO3 protein was stained positively in the cytoplasm of tumor cells and epithelial cells in many normal tissues.Conclusion:The polyclonal antibody against AGO3 protein has been achieved successfully,and it provides an efficient tool for further studying the roles of AGO3 in the pathway of miRNA and RNA interference and in the pathogenesis of human disease.

Natural killer (NK) cells, as an essential part of innate immunity, can directly identify and kill tumor cells after being activated by the synergistic action of surface inhibitory receptors and activated receptors. It can secrete cytokines to recruit dendritic cells (DCs), induce DCs maturation and enhance adaptive immune response. It can target cancer stem cells (CSCs) and circulating tumor cells (CTCs) to inhibit cancer metastasis. NK cells have a unique inflammatory tendency, which can respond to cytokines and chemokines released from tumor sites and migrate to tumor sites, making them occupy an important advantage in cancer targeted therapy. The research on cancer targeted therapy of NK cells as drug delivery carriers, NK cell membrane-coated biomimetic nanoparticles, and NK cell extracellular vesicles (NKEVs) has attracted more and more attention. The article will focus on the mechanism of NK cells inhibiting cancer, and summarize the research progress of cancer targeted therapy of NK cells.

Objective To prepare polyclonal antibodies against human PIWIL and to identify their property and tissue distribution of PIWIL.Methods PIWIL polypeptide was synthesized and conjugated to Keyhole limpet hemocyanin(KLH) as an immunogen.Then PIWIL-KLH conjugations were injected into rabbits subcutaneously to produce polyclonal antibodies.The specificity and sensitivity of antibodies were identified by ELISA and Western blot after purification by affinity chromatography.PIWIL were then stained on the tissue chip to study their distribution.Results Rabbit antibodies against PIWIL were prepared after injection of PIWIL-KLH conjugation.These antibodies specially recognized PIWIL peptides.Expression of PIWIL was found in the cytoplasm of epithelia cells of varied normal tissues and tumor tissues.Conclusion The successful preparation of the polyclonal antibody against PIWIL will provide an efficient reagent for further study of its role in the pathway of miRNA and RNA interference and in the pathogenesis of human disease.

Objective To investigate the mechanism and effect of the spinal cord outlet of the skull base on Chiari Ⅰ malformation with syringomyelia.Methods The cervical spinal cord stem angle (Anbc),slope angle of cervical vertebra (Ansc) of Chiari Ⅰ malformation were measured.In foramen magnum (Llf) and anterior vertebral canal level (Laf),spinal canal(Ac),spinal cord (As) and inferior hernia area (Ah) were measured.Angle,area and ratio were compared in Chiari Ⅰ malformation with syringomyelia,Chiari Ⅰ malformation without syringomyelia and normal control group.Results Ansc,Anbc-Ansc had significant differences among control group and Chiari Ⅰ malformation patients (all P＜0.001).In Llf,Laf,As had significant differences among three groups (all P＜0.05),further comparison of the two showed there were significant differences between Chiari Ⅰ malformation with syringomyelia patients and control group,Chiari Ⅰ malformation without syringomyelia patients and control group in Llf(all P＜0.05).In Llf,Laf,Ac in Chiari Ⅰ malformation with syringomyelia was smaller than control group (P＜0.05).Ah in Llf,Lafand Lh in Llf had no statistical significant difference between Chiari Ⅰ malformation with and without syringomyelia patients (all P＞0.05).In Llf,Laf,As/Ac had statistical significant difference among Chiari Ⅰ malformation with and without syringomyelia patients,control group (all P＜0.001),further comparison of the two showed As/Ac in Llf had statistical significance difference between Chiari Ⅰ malformation with syringomyelia patients and control group (P＜0.05),As/Ac in La had statistical significance difference between Chiari Ⅰ malformation with syringomyelia patients and control group,between Chiari Ⅰ malformation without syringomyelia patients and control group (all P＜0.05),Conclusion The cervical spinal cord,Ansc reducing,narrow vertebral proportion increase are important factors to promote Chiari Ⅰ malformation syringomyelia.

Objective To study the high-field MRI findings of gastrointestinal stromal tumors(GIST), investigate the diagnostic value of the tumor.Methods The image findings and clinical date of 45 cases with GIST proved by pathology were analyzed retrospectively.Results In 45 cases of GIST,tumors were located in the sacroanterior 2 cases, in the perineum 5 cases, in the small bowel 5 cases, in the rectum 10 cases and in the stomach 23 cases.Of 45 cases, there were 11 cases of recurrence,and 34 cases of new-onset.High-field MRI showed the tumors with inhomogeneous signal intensity, the small lesions with even homogeneous signals, the larger with non-uniform signalwhich may be cystic degeneration, necrosis, hemorrhage.After enhanced scan,the solid part of the tumor were abnormal enhancement,necrosis andcystic degeneration no enhancement.Clear or not clear of tumor boundaries andadjacent tissue pressure were prone to metastasis or intraperitoneal planting.Pathology results showed: the tumors cut surface was gray, gray-red, visible envelope.The microscopic morphology was varied, ranging from the number of spindle cells.Immunohistochemical findings were as following: CD34(+) in 43 cases;CD117(+) in 40 cases;Desmin(+) in 18 cases;Vimentin(+) in 15 cases.Conclusion High-field MRI can accurately position gastrointestinal stromal tumor, and has great value for its diagnosis and differential diagnosis.

OBJECTIVETo observe the changes of urinary deoxypyridinoline crosslink/creatinine (UDpd/Cr) in rats after OVX and intervention by estrogen and bisphosphonate and investigate the possible application of deoxypyridinoline in osteoporosis diagnosis and treatment.

METHODS40 female 6-month-old virginal Wistar rats were divided into 5 groups, ovariectomized or sham ovariectomized. (1) Ovxb (n = 8): sacrificed at 6 weeks after OVX; (2) Sham (n = 8): sham ovariectomized; (3) Ovxe (n = 8): sacrificed at 14 weeks after OVX; (4) O + E (n = 9):OVX + 17 beta estradiol [20 micrograms/(kg.d) ih]; (5) O + C (n = 7):OVX + cimadronate [0.2 mg/(kg.d)]; Treatment started 6 weeks after OVX and lasted 8 weeks. Rats in group 2-5 were sacrificed at 14 weeks after OVX. Urinary and serum biochemical parameters were measured, pQCT scanning of femur, bone biomechanical test in femur were determined.

RESULTSOVX resulted in increasing of UDpd/Cr 133.3% (P < 0.01). The ratio of UCa/Cr also increased in OVX groups but without any significant compared with Sham (P > 0.05). UDpd/Cr were reduced by 54.6% and 51.8% (P < 0.01) in O + E, O + C group respectively compared with Ovxe. The significant negative correlationships were found between UDpd/Cr and bone mass, BMD and biomechanic characteristics.

CONCLUSIONSUDpd/Cr ratio is a sensitive bone resorption marker, a marked changes were observed when the rats ovariectomized or treated with estradiol and cimadronate. There were best correlation between UDpd/Cr and bone mineral density and bone biomechanic characteristics. It is fair to apply UDpd/Cr ratio for osteoporosis diagnosis and treatment.

Amino Acids ; urine ; Animals ; Bone Density ; Creatinine ; urine ; Diphosphonates ; therapeutic use ; Estradiol ; therapeutic use ; Female ; Osteoporosis ; drug therapy ; urine ; Ovariectomy ; Rats ; Rats, Wistar