As a new hypnotic, zopiclone is widely used in clinical treatment. There are many methods for determination of zopiclone, including spectrophotometry, chromatography and chromatography mass spectrum, etc. Present paper reviews different kinds of biological samples associated with zopiclone, ex-traction and purification methods, and determination and analysis methods, which aims to provide refer-ences for the relevant research and practice.

Objective: The current study was designed to distinguish Phase G0/G1， S， G2/M cell groups in the whole cell cycle by DNA flow cytometry. Method: The authors developed a multiparameter flow cytometry of cyclin E＋ A/DNA for MOLT-4. Result: The data showed that the method could distinguish Phase G0, early G1, late G1, S, G2,and M cells. The method could be used in molecular cell biology, especially in cell kinetic study.

OBJECTIVETo investigate the modulation of pilose antler extract (PAE) on rat osteogenic cells UMR-106 in vitro.

METHODComponent P2 of PAE was isolated by Sephacryl S-200HR gel filtration chromatography. The proliferative effects of P2 and other components isolated by Sephacryl S-200HR on UMR-106 cells were investigated by MTT assay.

RESULTThe P2 could significantly increase the proliferation rate of osteogenic cells. When the protein concentration of P2 was between 0.972 mg x L(-1) and 97.2 mg x L(-1), it could inhibit the proliferation of UMR-106 cells. But while the concentration was equal to or greater than 97.2 mg x L(-1), the P2 could increase the proliferation rate of cells, especially 477.92% at 9.72 g x L(-1), which was approximated to 499.62% of PAE. The molecular weight of the P2 was about 59 kDa determined by SDS-PAGE. On the other hand, inhibition was also observed in the sample of the P3, P4 and P5.

CONCLUSIONThose regulative factors in PAE which have different molecular weight can affect the proliferation of UMR-106 cells two-wayly. And this adjustment also relies on the dose of those factors. This finding may help us to understand the possible mechanism of Chinese traditional medicine from animal materials.

Animals ; Antlers ; chemistry ; Bone Neoplasms ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Deer ; Materia Medica ; isolation & purification ; pharmacology ; Osteosarcoma ; pathology ; Rats ; Tissue Extracts ; isolation & purification ; pharmacology

OBJECTIVETo analyze the existence and the dynamic cell frequencies of human cells in goats transplanted in utero with human hematopoietic stem cell (hHSC) by using fluorescence in situ hybridization (FISH) technique.

METHODSInterphase FISH (IFISH) with human-specific 17-chromosome satellite DNA and/or human-specific Y-chromosome satellite DNA as probes was performed to analyze the presence and proportions of human cells in 13 transplanted goats. Samples were peripheral blood cells, bone marrow smears and liver touch imprint preparations.

RESULTSOf the 13 transplanted goats, eleven were identified to present human cells. Among them, two goats transplanted with human male HSC were found to have human male cells. The results demonstrated that these transplanted goats were human/goat HSC xenogeneic chimeras. Human cell frequencies decreased with the goat age (months), but the longest survival reached 21 months. During the detected life periods of goats, human cell frequencies in peripheral blood, bone marrow and liver tissues were less than 1@1000, but local human cell frequencies of 207.92@1000 and 392.41@1000 were detected in the liver tissues of 2 transplanted goats.

CONCLUSIONSThe existence and long-term survival of human cells in transplanted goats detected by FISH indicated that goats were appropriate recipients for hHSC in utero transplantation. The lower human cell frequencies in blood and bone marrow, and the higher local human cell frequencies in liver tissues suggested that the microenvironment of goat liver tissues might favor the survival, proliferation and differentiation of human cells.

Animals ; Female ; Goats ; Hematopoietic Stem Cell Transplantation ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Transplantation, Heterologous ; Uterus ; surgery

OBJECTIVEHigh-sensitivity C-reactive protein (hs-CRP) may predict the development of type 2 diabetes mellitus (T2DM), metabolic syndrome (MS) and cardiovascular diseases (CVD) in adult, but few reports on relevant studies in children are available. The present study aimed to understand possible correlation between serum hs-CRP levels and some factors of obese children and adolescents with or without impaired glycometabolism.

METHODSSeventy obese children and adolescents (age 8 - 17 years) and 30 non-obese healthy controls (group 1, 20 boys and 10 girls, mean age 12.6 years) were enrolled into this study. The obese individuals were subdivided into two groups according to the results of oral glucose tolerance test: the obese subjects without IGR (group 2, 54 cases, 43 boys and 11 girls, mean age 11.3 years) and the obese subjects with impaired glycometabolism (group 3, 16 cases, 8 boys and 8 girls, mean age 12.8 years). The levels of serum parameters including hs-CRP, glucose, lipid, insulin, C-peptide and whole blood HbA1c were determined. SPSS 10.0 was used for statistical analysis.

RESULTS(1) There was significant increase of serum hs-CRP level in obese children and adolescents, the median was 2.44 (0.01 - 14.6) mg/L; the level of control group was 0.1 (0.01 - 2.1) mg/L. (2) Some of the following parameters, such as fasting plasma glucose (FPG), triglyceride (TG), fasting insulin (FINS), C-peptide (Cp) and insulin resistance index (IRI), were found increased in group 2 and 3 as compared to group 1. When FPG and TG were still in normal range in group 2, the levels of hs-CRP and IRI were significantly higher than those in group 1, the level of hs-CRP was 2.4 (0.01 - 9.0) mg/L. While FPG and TG were abnormal in group 3, the level of hs-CRP was 2.6 (0.1 - 14.6) mg/L, but the difference had no statistical significance. (3) Pearson correlation analysis showed that there was a moderate correlation between serum hs-CRP and BMI (r = 0.414, P = 0.000). There was a low correlation between hs-CRP and waist circumference, hip circumference and waist to hip ratio (WHR). The correlation of serum hs-CRP with blood pressure, TG, cholesterol, high density lipoprotein-cholesterol (HDL-C), HbA1c, FPG, FINS and Cp had no significant deviation. (4) Multiple linear regression analysis showed that body mass index (BMI) was the only indicator which had correlation with hs-CRP.

CONCLUSION(1) There may be a chronic low-grade inflammation and insulin resistance in obese children. (2) The level of hs-CRP might be independently correlated with BMI in children. (3) Hs-CRP and IRI elevated before FPG and TG did, which may suggest that the low-grade inflammation and insulin resistance may be a pathogenic base of DM rather than the outcome of it. (4) The elevation of hs-CRP may help predict impaired glucose and lipid metabolism.

Adolescent ; Blood Glucose ; analysis ; Body Mass Index ; C-Reactive Protein ; analysis ; Case-Control Studies ; Child ; Female ; Glucose Tolerance Test ; Humans ; Insulin Resistance ; Lipoproteins, HDL ; blood ; Male ; Obesity ; blood ; metabolism ; Triglycerides ; blood ; Waist Circumference ; Waist-Hip Ratio

Objective　To investigate the expression of HGF and TGF-α and their receptor, Met (HGF receptor) and EGFR (TGF-αreceptor) mRNA, in the regenerative liver/hepatocytes after 70% partial hepatectomy (70% PH) in noncirrhotic biliary obstruction rats. Methods　Wistar rats were divided randomly into N-PH group, BDO-RBF-PH group and BDO-RBF group. The expression of HGF/Met mRNA and TGF-α/EGFR mRNA was measured by RT-PCR in the liver/hepatocytes at the time point of 0, 6, 12, 24, 48 and 72 h after 70% PH or RBF. Results　In N-PH group, the expression of HGF/Met mRNA increased sharply and peaked at 6 h, and maintained at a high level until 24 h after 70% PH. In BDO-RBF-PH group however, the expression of HGF/Met mRNA increased slowly and peaked at 12 h after 70% PH. The peak level was lower in BDO-RBF-PH group than in N-PH one. The expression of TGF-α/EGFR mRNA increased sharply and peaked at 24 h after 70% PH in N-PH group. However, the expression of TGF-α/EGFR mRNA elevated slowly and peaked at 48 h after 70% PH in BDO-RBF-PH group with a lower peak level than that in N-PH group. Conclusion　The expression of HGF/Met mRNA and TGF-α/EGFR mRNA in the regenerative liver/hepatocytes after 70% PH decreases significantly in noncirrhotic biliary duct obstruction rats. There is a tendency that the expression of HGF mRNA and TGF-α mRNA is less than Met mRNA and EGFR mRNA.

Objective To study the characteristics of a new neuron-specific marker in the cerebral cortex and its application method.Methods According to immunofluorescence principles,the neurons in the cerebral cortex cultured in vitro were stained by fluorescein isothiocyanate (FITC)-labeled Tet1; and FITC-Tet1 with NSE or GFAP was employed to determine whether the neurons can be marked and whether they can be distinguished from astrocytes; the conditions of application of FITC-Tet1 were quantified.The HEK and C6 cell lines were performed the above experiments as controls.Results The final concentration was the key factor,which indicated that when the FITC-Tet1 was about 30 μg/mL,the neuron marking ability was the best,and the astrocytes,HEK and C6 were not stained.Neither high (100μg/mL) or low (10μg/mL) levels of FITC-Tet1 were suitableforstainingneurons.Conclusion Tet1 can specifically combine with the neuron in the cerebral cortex,which may make it to be a new staining method or a targeting molecule to neurons in the central nervous system.

Objective To observe the relationship between serum and monocyte-derivedmacrophages secreted adipocyte fatty acid binding protein(A-FABP), adiponectin(or A-FABP/adiponectin ratio)and coronary artery disease. Methods Three hundred and forty subjects underwent coronary angiography(CAG)were classified into CAD group(n = 211)and non-CAD group(n = 129)according to the CAG results. The severity of coronary artery stenosis was assessed by the numbers of involved coronary artery branches and the sum of the Gensini scores. Fasting venous blood was collected from all subjects and peripheral monocytes were isolated from 20 subjects(10 selected from each group with age-, gender-, and BMI-matched). Peripheral blood monocytes were obtained and stimulated into macrophages with PMA, cell culture supernatant was collected. The concentration of serum/supernatant A-FABP and adiponectin levels were assayed by enzyme-linked immunosorbent assays. Results(1)A-FABP levels tendend to be higher in CAD patients compared to non-CAD subjects[18. 3(13.2,22. 8)μg/L vs. 16. 4(13.5,20. 4)μg/L,P = 0. 088]. The concentration of adiponectin in CAD group was significantly lower than those in non-CAD group[13.9(9.8,17.1)mg/L vs. 19.7(14.5,27.6)mg/L,P ＜0.05].(2)The A-FABP levels increased and the adiponectin levels decreased as the number of stenotic vessels increased. Gensini scores were positively correlated with serum A-FABP(r = 0. 120, P = 0.043)and inversely correlated with adiponectin(r = - 0. 405, P = 0. 007).(3)The difference in A-FABP/adiponectin ratio was moreprominent between subjects with CAD and subjects without CAD[(1.51 ±0. 79)μg/mg vs.(0. 89 ±0. 30)μg/mg, P ＜ 0. 01]and there was a stronger positive correlation of Gensini score to A-FABP/adiponectin ratio(r =0. 531, P =0. 000).(4)Monocyte-derived-macrophages from patients with CAD had higher A-FABP/adiponectin ratio than that in patients without CAD[(0. 51 ± 0. 19)μg/mg vs.(0. 36 ± 0. 11)μg/mg, P ＜ 0. 05]. Conclusions Increased levels of serum A-FABP and reduced levels of adiponectin in CAD patients serves as a novel biomarker for the severity of the coronary stenosis. A-FABP/adiponectin ratio is superior to A-FABP or adiponectin alone on predicting CAD risks.

Objective To research the effects of chronic suppurative otitis media on bone conduction threshold in old patients. Methods The files of patients with unilateral chronic otitis media were retrospective analyzed, who were all oder than 60 years, who were inpatient in our department since January 2005 to March 2009. Conventional puretone audiometry test was carried out. Bone conduction thresholds were calculated for frequencies of 0. 5, 1,2, and 4 kHz, with comparison between the ear with chronic otitis media and controlateral ear. Thresholds were examined separately for each frequency. Results The bone conduction threshold for the normal side was lower than those for the ear with chronic otitis media. The threshold shift was statistically significant for each frequency(P ＜0. 01 ). There were no diverences between the groups when analyzed for the presence of cholesteatoma except at 2 kHz frequencies ( Z = - 1. 975,P = 0. 048). There were diverences between the groups when analyzed for an interruption of the ossicular chain only at 2 kHz frequencies ( Z = - 2. 721, P = 0. 007 ). There were diverences between the groups when the duration of middle ear disease was not same at 1 kHz and 2 kHz frequencies( Z value were - 2. 877,-2. 624, P＜0. 01, respectively). Conclusions This study shows that chronic otitis media can enhance bone conduction threshold for old patients. All measures for early cure should be considered as early as possible in oder patients with chronic otitis media to prevent advance of sensorineural hearing loss.

The clinical value of transvaginal ultrasound in clinical surgical treatment of cesarean scar pregnancy (CSP) was evaluated.The clinical data of 104 patients with CSP admitted at Tongji Hospital from 2013 to 2016 were collected and analyzed retrospectively,including the patients' age,gestational age,the size of gestational sac or uneven mass,the uterine scar thickness,β human chorionic gonadotropin (hCG) levels and so on.Of these 104 cases,30 cases were subjected to laparotomy,29 cases to laparoscopy,27 cases to hysteroscopy,16 cases to ultrasound-guided uterine curettage,and 2 cases to conservative treatment.The transvaginal ultrasound showed that uterine scar thickness and gestational sac or uneven mass size had significant difference (P＜0.05) among different surgical methods by comparatively analyzing the patients' data.It was suggested that transvaginal ultrasound may provide the valuable reference for choosing clinical surgical procedures for CSP.